bims-bicyki Biomed News
on Bicaudal-C1 and interactors in cystic kidney disease
Issue of 2021–07–04
forty-four papers selected by
Céline Gagnieux, École Polytechnique Fédérale de Lausanne
  1. Gender-Dependent Phenotype in Polycystic Kidney Disease Is Determined by Differential Intracellular Ca2+ Signals.
  2. PKD1-Associated Arachnoid Cysts in Autosomal Dominant Polycystic Kidney Disease.
  3. Primary results of the randomized trial of metformin administration in polycystic kidney disease (TAME PKD).
  4. Coregulation Analysis of Mechanistic Biomarkers in Autosomal Dominant Polycystic Kidney Disease.
  5. Molecular Pathophysiology of Autosomal Recessive Polycystic Kidney Disease.
  6. Biallelic Mutations in DNAJB11 are Associated with Prenatal Polycystic Kidney Disease in a Turkish Family.
  7. Aquaporin 2 regulation: implications for water balance and polycystic kidney diseases.
  8. The effects of intrinsic apoptosis on cystogenesis in PKD1-deficient ADPKD pig model.
  9. DEAD-Box RNA Helicases in Cell Cycle Control and Clinical Therapy.
  10. Primary prevention of cardiovascular disease events with renin-angiotensin system blockade in autosomal dominant polycystic kidney disease dialysis patients: A nationwide cohort study.
  11. The ciliary impact of nonciliary gene mutations.
  12. Identification of Cilia in Different Mouse Tissues.
  13. Primary Cilium Is Involved in Stem Cell Differentiation and Renewal through the Regulation of Multiple Signaling Pathways.
  14. Increased expression of yes-associated protein/YAP and transcriptional coactivator with PDZ-binding motif/TAZ activates intestinal fibroblasts to promote intestinal obstruction in Crohn's disease.
  15. Blood pressure in children with renal cysts and diabetes syndrome.
  16. Oncogenic FGFR Fusions Produce Centrosome and Cilia Defects by Ectopic Signaling.
  17. Thrombospondin-1 mediates Rho-kinase inhibitor-induced increase in outflow-facility.
  18. Cyclic AMP does double duty to support parallel signaling in primary cilium and cytosol.
  19. Immortalization and Characterization of Rat Lingual Keratinocytes in a High-Calcium and Feeder-Free Culture System Using ROCK Inhibitor Y-27632.
  20. Hypothalamic primary cilium: A hub for metabolic homeostasis.
  21. [Paramecium, a model organism to study ciliogenesis and ciliopathies].
  22. Effect of Rho-Associated Kinase Inhibitor and Mesenchymal Stem Cell-Derived Conditioned Medium on Corneal Endothelial Cell Senescence and Proliferation.
  23. Centriolar Protein C2cd3 Is Required for Craniofacial Development.
  24. Multifaceted functions of Rab23 on primary cilium- and Hedgehog signaling-mediated cerebellar granule cell proliferation.
  25. Etoposide Triggers Cellular Senescence by Inducing Multiple Centrosomes and Primary Cilia in Adrenocortical Tumor Cells.
  26. Trace Amine-Associated Receptor 1 Trafficking to Cilia of Thyroid Epithelial Cells.
  27. ROCK Inhibition as Potential Target for Treatment of Pulmonary Hypertension.
  28. UA-Zero as a Uranyl Acetate Replacement When Diagnosing Primary Ciliary Dyskinesia by Transmission Electron Microscopy.
  29. Regulation of the Actin Cytoskeleton via Rho GTPase Signalling in Dictyostelium and Mammalian Cells: A Parallel Slalom.
  30. Microfluidic chip grafted with integrin tension sensors for evaluating the effects of flowing shear stress and ROCK inhibitor on platelets.
  31. Gene and epigenetic editing in the treatment of primary ciliopathies.
  32. MRTF: Basic Biology and Role in Kidney Disease.
  33. Exposure to 16 Hz Pulsed Electromagnetic Fields Protect the Structural Integrity of Primary Cilia and Associated TGF-β Signaling in Osteoprogenitor Cells Harmed by Cigarette Smoke.
  34. Early perturbation of Wnt signaling reveals patterning and invagination-evagination control points in molar tooth development.
  35. New Structural Perspectives in G Protein-Coupled Receptor-Mediated Src Family Kinase Activation.
  36. CEP104 and CEP290; Genes with Ciliary Functions Cause Intellectual Disability in Multiple Families.
  37. MITF Promotes Cell Growth, Migration and Invasion in Clear Cell Renal Cell Carcinoma by Activating the RhoA/YAP Signal Pathway.
  38. Evaluation of a Clinical Index as a Predictive Tool for Primary Ciliary Dyskinesia.
  39. WIP, YAP/TAZ and Actin Connections Orchestrate Development and Transformation in the Central Nervous System.
  40. Concurrent YAP/TAZ and SMAD signaling mediate vocal fold fibrosis.
  41. PRP4 Promotes Skin Cancer by Inhibiting Production of Melanin, Blocking Influx of Extracellular Calcium, and Remodeling Cell Actin Cytoskeleton.
  42. Centriole and Golgi microtubule nucleation are dispensable for the migration of human neutrophil-like cells.
  43. [Erratum] Regulation and mechanism of YAP/TAZ in the mechanical microenvironment of stem cells (Review).
  44. Fluid flow-induced left-right asymmetric decay of Dand5 mRNA in the mouse embryo requires a Bicc1-Ccr4 RNA degradation complex.
  1. Int J Mol Sci. 2021 Jun 02. pii: 6019. [Epub ahead of print]22(11):
    Gender-Dependent Phenotype in Polycystic Kidney Disease Is Determined by Differential Intracellular Ca2+ Signals.
    Khaoula Talbi, Inês Cabrita, Rainer Schreiber, Karl Kunzelmann.
      Autosomal dominant polycystic kidney disease (ADPKD) is caused by loss of function of PKD1 (polycystin 1) or PKD2 (polycystin 2). The Ca2+-activated Cl- channel TMEM16A has a central role in ADPKD. Expression and function of TMEM16A is upregulated in ADPKD which causes enhanced intracellular Ca2+ signaling, cell proliferation, and ion secretion. We analyzed kidneys from Pkd1 knockout mice and found a more pronounced phenotype in males compared to females, despite similar levels of expression for renal tubular TMEM16A. Cell proliferation, which is known to be enhanced with loss of Pkd1-/-, was larger in male when compared to female Pkd1-/- cells. This was paralleled by higher basal intracellular Ca2+ concentrations in primary renal epithelial cells isolated from Pkd1-/- males. The results suggest enhanced intracellular Ca2+ levels contributing to augmented cell proliferation and cyst development in male kidneys. Enhanced resting Ca2+ also caused larger basal chloride currents in male primary cells, as detected in patch clamp recordings. Incubation of mouse primary cells, mCCDcl1 collecting duct cells or M1 collecting duct cells with dihydrotestosterone (DHT) enhanced basal Ca2+ levels and increased basal and ATP-stimulated TMEM16A chloride currents. Taken together, the more severe cystic phenotype in males is likely to be caused by enhanced cell proliferation, possibly due to enhanced basal and ATP-induced intracellular Ca2+ levels, leading to enhanced TMEM16A currents. Augmented Ca2+ signaling is possibly due to enhanced expression of Ca2+ transporting/regulating proteins.
    Keywords:  ADPKD; CFTR; TMEM16A; androgen; estrogen; polycystic kidneys
    DOI:  https://doi.org/10.3390/ijms22116019
  2. J Stroke Cerebrovasc Dis. 2021 Jun 24. pii: S1052-3057(21)00346-3. [Epub ahead of print]30(9): 105943
    PKD1-Associated Arachnoid Cysts in Autosomal Dominant Polycystic Kidney Disease.
    Kaori Shigemori, Eiji Higashihara, Masayuki Itoh, Hiroki Yoshida, Kouji Yamamoto, Kikuo Nutahara, Yoshiaki Shiokawa, Shinya Kaname, Mitsuhiro Tambo, Tsuyoshi Yamaguchi, Satoru Taguchi, Tatsuya Yoshioka, Kenichi Yokoyama, Hiroshi Fukuhara.
       OBJECTIVES: the prevalence of intracranial aneurysms and arachnoid cysts is higher in patients with autosomal dominant polycystic kidney disease (ADPKD) than in the general population. A genotype correlation was reported for intracranial aneurysms, but it is unclear for arachnoid cysts. Therefore, the genotype correlation with intracranial aneurysms and arachnoid cysts was investigated in ADPKD.
    MATERIALS AND METHODS: intracranial aneurysms and arachnoid cysts were screened by magnetic resonance imaging (MRI), and PKD genotypes were examined using next-generation sequencing for 169 patients with ADPKD.
    RESULTS: PKD1-, PKD2- and no-mutation were identified in 137, 24 and 8 patients, respectively. Intracranial aneurysms and arachnoid cysts were found in 34 and 25 patients, respectively, with no significant difference in frequency. Genotype, sex, estimated glomerular filtration rate and age at ADPKD diagnosis significantly affected the age at brain MRI. The proportional hazard risk analyzed using the age at brain MRI adjusted by these four variables was 5.0-times higher in the PKD1 group than in the PKD2 group for arachnoid cysts (P = 0.0357), but it was not different for intracranial aneurysms (P = 0.1605). Arachnoid cysts were diagnosed earlier in the PKD1 group than in the PKD2 group (54.8 vs 67.7 years, P = 0.0231), but no difference was found for intracranial aneurysms (P = 0.4738) by Kaplan-Meier analysis.
    CONCLUSIONS: this study demonstrated the correlation between arachnoid cysts and PKD1 mutation. The reported association of arachnoid cysts with advanced renal disease may be due to the common correlation of these factors with PKD1 mutation.
    Keywords:  Arachnoid cysts; Autosomal dominant polycystic kidney disease (ADPKD); Genotype; Intracranial aneurysms; Intracranial vascular event
    DOI:  https://doi.org/10.1016/j.jstrokecerebrovasdis.2021.105943
  3. Kidney Int. 2021 Jun 26. pii: S0085-2538(21)00601-3. [Epub ahead of print]
    Primary results of the randomized trial of metformin administration in polycystic kidney disease (TAME PKD).
    Ronald D Perrone, Kaleab Z Abebe, Terry Watnick, Andrew Althouse, Kenneth R Hallows, Christina M Lalama, Dana C Miskulin, Stephen L Seliger, Cheng Tao, Peter C Harris, Kyongtae Ty Bae.
      Autosomal dominant polycystic kidney disease (ADPKD) is characterized by growth of kidney cysts and glomerular filtration rate (GFR) decline. Metformin was found to impact cystogenesis in preclinical models of polycystic disease, is generally considered safe and may be a promising candidate for clinical investigation in ADPKD. In this phase 2 two-year trial, we randomly assigned 97 patients, 18-60 years of age, with ADPKD and estimated GFR over 50 ml/min/1.73 m2, in a 1:1 ratio to receive metformin or placebo twice daily. Primary outcomes were medication safety and tolerability. Secondary outcomes included estimated GFR decline, and total kidney volume growth. Thirty-eight metformin and 39 placebo participants still received study product at 24-months. Twenty-one participants in the metformin arm reduced drug dose due to inability to tolerate, compared with 14 in the placebo arm (not significant). Proportions of participants experiencing serious adverse events was similar between the groups. The Gastrointestinal Symptoms Rating Scale score was low at baseline and did not significantly change over time. The annual change for estimated GFR was -1.71 with metformin and -3.07 ml/min/1.73m2 per year with placebo (mean difference 1.37 {-0.70, 3.44} ml/min/1.73m2), while mean annual percent change in height-adjusted total kidney volume was 3.87% in metformin and 2.16% per year in placebo, (mean difference 1.68% {-2.11, 5.62}). Thus, metformin in adults with ADPKD was found to be safe and tolerable while slightly reducing estimated GFR decline but not to a significant degree. Hence, evaluation of efficacy requires a larger trial, with sufficient power to detect differences in endpoints.
    Keywords:  ADPKD; clinical trial; eGFR; metformin; total kidney volume
    DOI:  https://doi.org/10.1016/j.kint.2021.06.013
  4. Int J Mol Sci. 2021 Jun 26. pii: 6885. [Epub ahead of print]22(13):
    Coregulation Analysis of Mechanistic Biomarkers in Autosomal Dominant Polycystic Kidney Disease.
    Johannes Leierer, Paul Perco, Benedikt Hofer, Susanne Eder, Alexander Dzien, Julia Kerschbaum, Michael Rudnicki, Gert Mayer.
      Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary kidney disorder leading to deterioration of kidney function and end stage kidney disease (ESKD). A number of molecular processes are dysregulated in ADPKD but the exact mechanism of disease progression is not fully understood. We measured protein biomarkers being linked to ADPKD-associated molecular processes via ELISA in urine and serum in a cohort of ADPKD patients as well as age, gender and eGFR matched CKD patients and healthy controls. ANOVA and t-tests were used to determine differences between cohorts. Spearman correlation coefficient analysis was performed to assess coregulation patterns of individual biomarkers and renal function. Urinary epidermal growth factor (EGF) and serum apelin (APLN) levels were significantly downregulated in ADPKD patients. Serum vascular endothelial growth factor alpha (VEGFA) and urinary angiotensinogen (AGT) were significantly upregulated in ADPKD patients as compared with healthy controls. Arginine vasopressin (AVP) was significantly upregulated in ADPKD patients as compared with CKD patients. Serum VEGFA and VIM concentrations were positively correlated and urinary EGF levels were negatively correlated with urinary AGT levels. Urinary EGF and AGT levels were furthermore significantly associated with estimated glomerular filtration rate (eGFR) in ADPKD patients. In summary, altered protein concentrations in body fluids of ADPKD patients were found for the mechanistic markers EGF, APLN, VEGFA, AGT, AVP, and VIM. In particular, the connection between EGF and AGT during progression of ADPKD warrants further investigation.
    Keywords:  EGFR signaling; angiogenesis; autosomal dominant polycystic kidney disease; mechanistic biomarkers
    DOI:  https://doi.org/10.3390/ijms22136885
  5. Int J Mol Sci. 2021 Jun 17. pii: 6523. [Epub ahead of print]22(12):
    Molecular Pathophysiology of Autosomal Recessive Polycystic Kidney Disease.
    Adrian Cordido, Marta Vizoso-Gonzalez, Miguel A Garcia-Gonzalez.
      Autosomal recessive polycystic kidney disease (ARPKD) is a rare disorder and one of the most severe forms of polycystic kidney disease, leading to end-stage renal disease (ESRD) in childhood. PKHD1 is the gene that is responsible for the vast majority of ARPKD. However, some cases have been related to a new gene that was recently identified (DZIP1L gene), as well as several ciliary genes that can mimic a ARPKD-like phenotypic spectrum. In addition, a number of molecular pathways involved in the ARPKD pathogenesis and progression were elucidated using cellular and animal models. However, the function of the ARPKD proteins and the molecular mechanism of the disease currently remain incompletely understood. Here, we review the clinics, treatment, genetics, and molecular basis of ARPKD, highlighting the most recent findings in the field.
    Keywords:  ARPKD; cyst; nephrology; rare monogenic disease
    DOI:  https://doi.org/10.3390/ijms22126523
  6. Mol Syndromol. 2021 Jun;12(3): 179-185
    Biallelic Mutations in DNAJB11 are Associated with Prenatal Polycystic Kidney Disease in a Turkish Family.
    Esra Arslan Ateş, Ayberk Turkyilmaz, Kenan Delil, Ceren Alavanda, Mehmet Ali Söylemez, Bilgen Bilge Geçkinli, Pinar Ata, Ahmet Arman.
      Polycystic kidney disease (PKD) is a life-threatening condition resulting in end-stage renal disease. Two major forms of PKD are defined according to the inheritance pattern. Autosomal dominant PKD (ADPKD) is characterized by renal cysts, where nearly half of the patients suffers from renal failure in the 7th decade of life. Autosomal recessive PKD (ARPKD) is a rarer and more severe form presenting in childhood. Whole-exome sequencing (WES) analyses was performed to investigate molecular causes of the disease in the fetus. In this study, we present 2 fetuses prenatally diagnosed with PKD in a consanguineous family. WES analysis of the second fetus revealed a homozygous variant (c.740+1G>A) in DNAJB11 which is related to ADPKD. This study reveals that DNAJB11 biallelic mutations may cause an antenatal severe form of ARPKD and contributes to understanding the DNAJB11-related ADPKD phenotype. The possibility of ARPKD due to biallelic mutations in ADPKD genes should be considered in genetic counseling.
    Keywords:  DNAJB11; Genetic counseling; Nephrology; Polycystic kidney disease; Whole-exome sequencing
    DOI:  https://doi.org/10.1159/000513611
  7. Nat Rev Nephrol. 2021 Jul 01.
    Aquaporin 2 regulation: implications for water balance and polycystic kidney diseases.
    Emma T B Olesen, Robert A Fenton.
      Targeting the collecting duct water channel aquaporin 2 (AQP2) to the plasma membrane is essential for the maintenance of mammalian water homeostasis. The vasopressin V2 receptor (V2R), which is a GS protein-coupled receptor that increases intracellular cAMP levels, has a major role in this targeting process. Although a rise in cAMP levels and activation of protein kinase A are involved in facilitating the actions of V2R, studies in knockout mice and cell models have suggested that cAMP signalling pathways are not an absolute requirement for V2R-mediated AQP2 trafficking to the plasma membrane. In addition, although AQP2 phosphorylation is a known prerequisite for V2R-mediated plasma membrane targeting, none of the known AQP2 phosphorylation events appears to be rate-limiting in this process, which suggests the involvement of other factors; cytoskeletal remodelling has also been implicated. Notably, several regulatory processes and signalling pathways involved in AQP2 trafficking also have a role in the pathophysiology of autosomal dominant polycystic kidney disease, although the role of AQP2 in cyst progression is unknown. Here, we highlight advances in the field of AQP2 regulation that might be exploited for the treatment of water balance disorders and provide a rationale for targeting these pathways in autosomal dominant polycystic kidney disease.
    DOI:  https://doi.org/10.1038/s41581-021-00447-x
  8. Gene. 2021 Jun 24. pii: S0378-1119(21)00387-5. [Epub ahead of print] 145792
    The effects of intrinsic apoptosis on cystogenesis in PKD1-deficient ADPKD pig model.
    Runming Wang, Wenya Li, Suhong Zhang, Ya Song, Haiting Dai, Tan Tan, Xiaoxiang Hu, Yiming Xing.
       BACKGROUND: Apoptosis is a form of cell death that plays a critical role in the maintenance of tissue homeostasis involving the development and elimination of unwanted cells. Dysregulation of apoptosis appears to be associated in the pathogenesis of many human diseases. Autosomal dominant polycystic kidney disease (ADPKD) is a common monogenetic disease and is mainly caused by mutations in PKD1. Previous studies proved that increased cell death occurred in ADPKD patients and animal models. However, the role of apoptosis in kidney cystogenesis is not clear.
    METHODS: In current study, due to the high similarities between human and pig, PKD1-deficient (PKD1+/-) pigs and PKD1-knockdown (PKD1KD) pig kidney epithelial cells were used to investigate the mechanisms of apoptosis in driving cystogenesis.
    RESULTS: In PKD1+/- pigs, increased intrinsic and extrinsic apoptosis were found at ages of 1 month and 3 months, whereas the autophagy and pyroptosis were not altered. Meanwhile, the intrinsic apoptosis was activated along with untouched extrinsic apoptosis in PKD1KD pig kidney cells. Thus, the intrinsic apoptosis played important roles in cystogenesis.
    CONCLUSIONS: This work provides detail analysis of the roles of different cell death types during cystogenesis in ADPKD pig model. The results suggested a potential new strategy for the diagnosis and treatment of ADPKD by targeting intrinsic apoptosis.
    Keywords:  ADPKD; Intrinsic apoptosis; PKD1; Pig
    DOI:  https://doi.org/10.1016/j.gene.2021.145792
  9. Cells. 2021 Jun 18. pii: 1540. [Epub ahead of print]10(6):
    DEAD-Box RNA Helicases in Cell Cycle Control and Clinical Therapy.
    Lu Zhang, Xiaogang Li.
      Cell cycle is regulated through numerous signaling pathways that determine whether cells will proliferate, remain quiescent, arrest, or undergo apoptosis. Abnormal cell cycle regulation has been linked to many diseases. Thus, there is an urgent need to understand the diverse molecular mechanisms of how the cell cycle is controlled. RNA helicases constitute a large family of proteins with functions in all aspects of RNA metabolism, including unwinding or annealing of RNA molecules to regulate pre-mRNA, rRNA and miRNA processing, clamping protein complexes on RNA, or remodeling ribonucleoprotein complexes, to regulate gene expression. RNA helicases also regulate the activity of specific proteins through direct interaction. Abnormal expression of RNA helicases has been associated with different diseases, including cancer, neurological disorders, aging, and autosomal dominant polycystic kidney disease (ADPKD) via regulation of a diverse range of cellular processes such as cell proliferation, cell cycle arrest, and apoptosis. Recent studies showed that RNA helicases participate in the regulation of the cell cycle progression at each cell cycle phase, including G1-S transition, S phase, G2-M transition, mitosis, and cytokinesis. In this review, we discuss the essential roles and mechanisms of RNA helicases in the regulation of the cell cycle at different phases. For that, RNA helicases provide a rich source of targets for the development of therapeutic or prophylactic drugs. We also discuss the different targeting strategies against RNA helicases, the different types of compounds explored, the proposed inhibitory mechanisms of the compounds on specific RNA helicases, and the therapeutic potential of these compounds in the treatment of various disorders.
    Keywords:  DDX3; DDX5; DEAD-box RNA helicases; cell cycle; treatment
    DOI:  https://doi.org/10.3390/cells10061540
  10. Medicine (Baltimore). 2021 Jul 02. 100(26): e26559
    Primary prevention of cardiovascular disease events with renin-angiotensin system blockade in autosomal dominant polycystic kidney disease dialysis patients: A nationwide cohort study.
    Chien-Lin Lu, Chien-Yu Lin, Lian-Yu Lin, Pau-Chung Chen, Cai-Mei Zheng, Kuo-Cheng Lu, Dong-Feng Yeih.
       ABSTRACT: Although renin-angiotensin system (RAS) blockade has been shown to reduce cardiovascular disease (CVD) in the general population and high-risk subjects, their protective effect in autosomal dominant polycystic kidney disease (ADPKD) patients under dialysis was still unknown. By using the database from 1995 to 2008 Taiwan National Health Insurance Research Database (Registry for Catastrophic Illnesses), we included 387 ADPKD patients who received dialysis therapy, aged ≥ 18 year-old, and with no evidence of CVD events in 1997 and 1998. We utilized Cox proportional hazards regression analysis and propensity score matching to evaluate adjusted hazard ratios for all-cause mortality and CVD events in users (n=231) and nonusers (n = 156) of an angiotensin-converting enzymes inhibitor (ACEI) / angiotensin II receptor blocker (ARB) during the 12 years of follow-up. All study subjects were followed up for more than 3 months. There was no significant difference between the ACEI/ARB treatment group and the control group in incident CVD events except ischemic stroke and transient ischemic accident (TIA). The results remain similar between groups before and after propensity score matching. Moreover, there was no significant difference in outcomes between ACEI/ARB treatment over 50% of follow-up period and without ACEI/ARB treatment after propensity score matching. This nationwide cohort study failed to prove the protective effects of long-term ACEI or ARB on incident CVD events among APKD dialysis patients. Further larger scale, multicenter and randomized control trials are warranted to show the causal association.
    DOI:  https://doi.org/10.1097/MD.0000000000026559
  11. Trends Cell Biol. 2021 Jun 25. pii: S0962-8924(21)00117-3. [Epub ahead of print]
    The ciliary impact of nonciliary gene mutations.
    Marta Lovera, Jens Lüders.
      Mutations in genes encoding centriolar or ciliary proteins cause diseases collectively known as 'ciliopathies'. Interestingly, the Human Phenotype Ontology database lists numerous disorders that display clinical features reminiscent of ciliopathies but do not involve defects in the centriole-cilium proteome. Instead, defects in different cellular compartments may impair cilia indirectly and cause additional, nonciliopathy phenotypes. This phenotypic heterogeneity, perhaps combined with the field's centriole-cilium-centric view, may have hindered the recognition of ciliary contributions. Identifying these diseases and dissecting how the underlying gene mutations impair cilia not only will add to our understanding of cilium assembly and function but also may open up new therapeutic avenues.
    Keywords:  centriole; cilia; ciliopathy; microtubules; signaling
    DOI:  https://doi.org/10.1016/j.tcb.2021.06.001
  12. Cells. 2021 Jun 29. pii: 1623. [Epub ahead of print]10(7):
    Identification of Cilia in Different Mouse Tissues.
    Xinhua Li, Shuting Yang, Vishwa Deepak, Zahra Chinipardaz, Shuying Yang.
      Cilia are microtubule-based hair-like organelles that extend from the cell surface. However, the existence and distribution of cilia in each organ and tissue at the postnatal stage in vivo remain largely unknown. In this study, we defined cilia distribution and arrangement and measured the ciliary lengths and the percentage of ciliated cells in different organs and tissues in vivo by using cilium dual reporter-expressing transgenic mice. Cilia were identified by the presence of ARL13B with an mCherry+ signal, and the cilium basal body was identified by the presence of Centrin2 with a GFP+ signal. Here, we provide in vivo evidence that chondrocytes and cells throughout bones have cilia. Most importantly, we reveal that: 1. primary cilia are present in hepatocytes; 2. no cilia but many centrioles are distributed on the apical cell surface in the gallbladder, intestine, and thyroid epithelia; 3. cilia on the cerebral cortex are well oriented, pointing to the center of the brain; 4. ARL13B+ inclusion is evident in the thyroid and islets of Langerhans; and 5. approximately 2% of cilia show irregular movement in nucleus pulposus extracellular fluid. This study reveals the existence and distribution of cilia and centrioles in different tissues and organs, and provides new insights for further comprehensive study of ciliary function in these organs and tissues.
    Keywords:  ARL13B; brain; centrin; nucleus pulposus; primary cilia
    DOI:  https://doi.org/10.3390/cells10071623
  13. Cells. 2021 Jun 08. pii: 1428. [Epub ahead of print]10(6):
    Primary Cilium Is Involved in Stem Cell Differentiation and Renewal through the Regulation of Multiple Signaling Pathways.
    Sila Yanardag, Elena N Pugacheva.
      Signaling networks guide stem cells during their lineage specification and terminal differentiation. Primary cilium, an antenna-like protrusion, directly or indirectly plays a significant role in this guidance. All stem cells characterized so far have primary cilia. They serve as entry- or check-points for various signaling events by controlling the signal transduction and stability. Thus, defects in the primary cilia formation or dynamics cause developmental and health problems, including but not limited to obesity, cardiovascular and renal anomalies, hearing and vision loss, and even cancers. In this review, we focus on the recent findings of how primary cilium controls various signaling pathways during stem cell differentiation and identify potential gaps in the field for future research.
    Keywords:  Notch; TGF; Wnt; cancer stem cells; differentiation; mTOR; primary cilia; signaling; stem cells
    DOI:  https://doi.org/10.3390/cells10061428
  14. EBioMedicine. 2021 Jun 26. pii: S2352-3964(21)00245-0. [Epub ahead of print]69 103452
    Increased expression of yes-associated protein/YAP and transcriptional coactivator with PDZ-binding motif/TAZ activates intestinal fibroblasts to promote intestinal obstruction in Crohn's disease.
    Weijun Ou, Weimin Xu, Fangyuan Liu, Yuegui Guo, Zhenyu Huang, Tienan Feng, Chen-Ying Liu, Peng Du.
       BACKGROUND: Intestinal obstruction caused by intestinal fibrosis is a common and serious complication of Crohn's disease (CD). Intestinal fibroblasts, the main effector cells mediating gastrointestinal fibrosis, are activated during chronic inflammation. However, the mechanism of fibroblast activation in CD has not been well elucidated.
    METHODS: Fibroblasts isolated from stenotic and nonstenotic intestines of CD patients were used for RNA sequencing. Immunohistochemical and immunofluorescent staining was performed to evaluate the correlation between intestinal fibrosis and YAP/TAZ expression in our CD cohort and a DSS-induced chronic colitis murine model. A Rho-associated coiled-coil-containing protein kinase 1 (ROCK1) inhibitor was used to explore the ROCK1-YAP/TAZ axis in intestinal fibroblasts in vitro and DSS-induced chronic colitis murine model in vivo.
    FINDINGS: The expression of YAP/TAZ was significantly upregulated in stenotic fibroblasts, which was associated with the YAP/TAZ target gene signature. YAP/TAZ knockdown suppressed the activation of intestinal fibroblasts. In intestinal fibroblasts, YAP/TAZ were activated by the Rho-ROCK1 signalling pathway. High YAP/TAZ expression was positively correlated with ROCK1 expression, which is a prognostic marker for intestinal obstruction in CD patients.
    INTERPRETATION: YAP/TAZ activation can lead to fibroblast activation and intestinal obstruction in CD. The effect of ROCK1 inhibitor on alleviating intestinal fibrosis is associated with YAP/TAZ inhibition. Targeted inhibition of YAP/TAZ in fibroblasts may be a potential therapeutic strategy to suppress intestinal fibrosis in CD.
    FUNDING: This work was supported by the National Key R&D Program of China (2019YFC1316002), the NSFC (81873547, 82073201, 81874177, 82000481) and the Shanghai Sailing Program (20YF1429400).
    Keywords:  Crohn's disease; ROCK1; TAZ; YAP; intestinal fibroblasts
    DOI:  https://doi.org/10.1016/j.ebiom.2021.103452
  15. Eur J Pediatr. 2021 Jun 26.
    Blood pressure in children with renal cysts and diabetes syndrome.
    Tomáš Seeman, Friederike Weigel, Kveta Blahova, Filip Fencl, Stepanka Pruhova, Katharina Hermes, Richard Klaus, Bärbel Lange-Sperandio, Veit Grote, Ulrike John-Kroegel.
      Cystic kidney diseases such as autosomal recessive or dominant polycystic kidney disease (ARPKD and ADPKD) are associated with high prevalence of arterial hypertension. On the contrary, studies on hypertension in children with renal cysts and diabetes (RCAD) syndrome caused by abnormalities in the HNF1B gene are rare. Therefore, the primary aim of our study was to investigate the prevalence of high blood pressure in children with RCAD syndrome due to HNF1B gene abnormalities and secondary to search for possible risk factors for development of high blood pressure. Data on all children with genetically proven RCAD syndrome from three pediatric nephrology tertiary centers were retrospectively reviewed (office blood pressure (BP), ambulatory blood pressure monitoring (ABPM), creatinine clearance, renal ultrasound, echocardiography, albuminuria/proteinuria). High blood pressure was defined as BP ≥ 95th percentile of the current ESH 2016 guidelines and/or by the use of antihypertensive drugs. Thirty-two children with RCAD syndrome were investigated. Three children received ACE inhibitors for hypertension and/or proteinuria. High blood pressure was diagnosed using office BP in 22% of the children (n = 7). In the 7 performed ABPM, 1 child (14%) was diagnosed with hypertension and one child with white-coat hypertension. Creatinine clearance, proteinuria, albuminuria, body mass index, enlargement, or hypodysplasia of the kidneys and prevalence of HNF1B-gene deletion or mutation were not significantly different between hypertensive and normotensive children.Conclusion: High blood pressure is present in 22% of children with RCAD syndrome. What is Known: • Arterial hypertension is a common complication in children with polycystic kidney diseases. What is New: • High office blood pressure is present in 22% and ambulatory hypertension in 14% of children with renal cyst and diabetes (RCAD) syndrome.
    Keywords:  Albuminuria; Ambulatory blood pressure monitoring; HNF1B; Hypertension; Proteinuria; Renal function
    DOI:  https://doi.org/10.1007/s00431-021-04165-1
  16. Cells. 2021 Jun 09. pii: 1445. [Epub ahead of print]10(6):
    Oncogenic FGFR Fusions Produce Centrosome and Cilia Defects by Ectopic Signaling.
    Alexandru Nita, Sara P Abraham, Pavel Krejci, Michaela Bosakova.
      A single primary cilium projects from most vertebrate cells to guide cell fate decisions. A growing list of signaling molecules is found to function through cilia and control ciliogenesis, including the fibroblast growth factor receptors (FGFR). Aberrant FGFR activity produces abnormal cilia with deregulated signaling, which contributes to pathogenesis of the FGFR-mediated genetic disorders. FGFR lesions are also found in cancer, raising a possibility of cilia involvement in the neoplastic transformation and tumor progression. Here, we focus on FGFR gene fusions, and discuss the possible mechanisms by which they function as oncogenic drivers. We show that a substantial portion of the FGFR fusion partners are proteins associated with the centrosome cycle, including organization of the mitotic spindle and ciliogenesis. The functions of centrosome proteins are often lost with the gene fusion, leading to haploinsufficiency that induces cilia loss and deregulated cell division. We speculate that this complements the ectopic FGFR activity and drives the FGFR fusion cancers.
    Keywords:  FGFR; FGFR fusion; cancer; centrosome; centrosome cycle; cilia; fibroblast growth factor receptor; neoplastic transformation; oncogenic driver; primary cilia
    DOI:  https://doi.org/10.3390/cells10061445
  17. J Cell Physiol. 2021 Jun 27.
    Thrombospondin-1 mediates Rho-kinase inhibitor-induced increase in outflow-facility.
    Sze-Wan Shan, Chi-Wai Do, Thomas Chuen Lam, Hoi-Lam Li, W Daniel Stamer, Chi-Ho To.
      Rho-kinase (ROCK) inhibitors, a novel class of anti-glaucoma agents, act by increasing the aqueous humor outflow through the conventional trabecular meshwork pathway. However, the downstream signaling consequences of the ROCK inhibitor are not completely understood. Our data show that Y39983, a selective ROCK inhibitor, could induce filamentous actin remodeling, reduced cell motility (as measured by cell migration), and transepithelial resistance in primary human TM (hTM) cells. After 2 days Y39983 treatment of hTM cells, a proteomic study identified 20 proteins whose expression was significantly altered. Pathway analysis of those proteins revealed the involvement of the p53 pathway, integrin signaling pathway, and cytoskeletal pathway regulation by Rho GTPase. Thrombospondin-1 (TSP1), a matricellular protein that is increased in glaucoma patients, was downregulated fivefold following Y39983 treatment. More importantly, both TSP1 antagonist leucine-serine-lysine-leucine (LSKL) and small interfering RNA (siRNA) reduced TSP1 gene and protein expressions as well as hTM cell migration. In the presence of Y39983, no further inhibition of cell migration resulted after LSKL and TSP1 siRNA knockdown. Likewise, LSKL triggered a dose-dependent increase in outflow facility in ex vivo mouse eyes, to a similar extent as Y39983 (83.8% increase by Y39983 vs. 71.2% increase by LSKL at 50 µM). There were no additive effects with simultaneous treatment with LSKL and Y39983, supporting the notion that the effects of ROCK inhibition were mediated by TSP1.
    Keywords:  ROCK inhibitor; glaucoma; outflow facility; thrombospondin-1; trabecular meshwork
    DOI:  https://doi.org/10.1002/jcp.30492
  18. Cell Calcium. 2021 Jun 24. pii: S0143-4160(21)00090-7. [Epub ahead of print]97 102436
    Cyclic AMP does double duty to support parallel signaling in primary cilium and cytosol.
    Danielle T Arena, Aldebaran M Hofer.
      The primary cilium maintains all of the necessary machinery to generate and interpret cAMP signals within its tiny volume, leading to the supposition that ciliary cAMP provides unique biological instructions separate from those derived from the rest of the cell body. A new paper by Truong et al. has used optogenetic and chemogenetic tricks to selectively manipulate cAMP signaling within the primary cilium. Their data show that ciliary but not cytosolic message preferentially regulates transcriptional activity via the hedgehog pathway leading to actions on zebrafish development. Computer modeling provides a rational explanation as to how the geometry of this organelle enables it to tune out cAMP signals from the cell body in order to pick up messages generated in the cilium.
    Keywords:  Cyclic AMP microdomain; DREADDs; Photoactivatable adenylyl cyclase bPAC; Primary cilium; Protein Kinase A; Zebrafish
    DOI:  https://doi.org/10.1016/j.ceca.2021.102436
  19. Int J Mol Sci. 2021 Jun 24. pii: 6782. [Epub ahead of print]22(13):
    Immortalization and Characterization of Rat Lingual Keratinocytes in a High-Calcium and Feeder-Free Culture System Using ROCK Inhibitor Y-27632.
    Zixing Chen, Wenmeng He, Thomas Chun Ning Leung, Hau Yin Chung.
      Cultured keratinocytes are desirable models for biological and medical studies. However, primary keratinocytes are difficult to maintain, and there has been little research on lingual keratinocyte culture. Here, we investigated the effect of Y-27632, a Rho kinase (ROCK) inhibitor, on the immortalization and characterization of cultured rat lingual keratinocyte (RLKs). Three Y-27632-supplemented media were screened for the cultivation of RLKs isolated from Sprague-Dawley rats. Phalloidin staining and TUNEL assay were applied to visualize cytoskeleton dynamics and cell apoptosis following Y-27632 removal. Label-free proteomics, RT-PCR, calcium imaging, and cytogenetic studies were conducted to characterize the cultured cells. Results showed that RLKs could be conditionally immortalized in a high-calcium medium in the absence of feeder cells, although they did not exhibit normal karyotypes. The removal of Y-27632 from the culture medium led to reversible cytoskeletal reorganization and nuclear enlargement without triggering apoptosis, and a total of 239 differentially expressed proteins were identified by proteomic analysis. Notably, RLKs derived from the non-taste epithelium expressed some molecular markers characteristic of taste bud cells, yet calcium imaging revealed that they rarely responded to tastants. Collectively, we established a high-calcium and feeder-free culture method for the long-term maintenance of RLKs. Our results shed some new light on the immortalization and differentiation of lingual keratinocytes.
    Keywords:  ROCK inhibitor; Y-27632; feeder-free; high calcium; immortalization; lingual keratinocytes; proteomics; taste bud cells
    DOI:  https://doi.org/10.3390/ijms22136782
  20. Exp Mol Med. 2021 Jul 01.
    Hypothalamic primary cilium: A hub for metabolic homeostasis.
    Dong Joo Yang, Jessica Hong, Ki Woo Kim.
      Obesity is a global health problem that is associated with adverse consequences such as the development of metabolic disorders, including cardiovascular disease, neurodegenerative disorders, and type 2 diabetes. A major cause of obesity is metabolic imbalance, which results from insufficient physical activity and excess energy intake. Understanding the pathogenesis of obesity, as well as other metabolic disorders, is important in the development of methods for prevention and therapy. The coordination of energy balance takes place in the hypothalamus, a major brain region that maintains body homeostasis. The primary cilium is an organelle that has recently received attention because of its role in controlling energy balance in the hypothalamus. Defects in proteins required for ciliary function and formation, both in humans and in mice, have been shown to cause various metabolic disorders. In this review, we provide an overview of the critical functions of primary cilia, particularly in hypothalamic areas, and briefly summarize the studies on the primary roles of cilia in specific neurons relating to metabolic homeostasis.
    DOI:  https://doi.org/10.1038/s12276-021-00644-5
  21. Med Sci (Paris). 2021 Jun-Jul;37(6-7):37(6-7): 632-638
    [Paramecium, a model organism to study ciliogenesis and ciliopathies].
    Khaled Bouhouche, Pierrick Le Borgne, Michel Lemullois, Anne-Marie Tassin.
      The cilium is a cell extension forming a distinct compartment of eukaryotic cell body with a complex and dynamic structure. This structure is highly conserved across species and ensures various functions as sensory and motility. In humans, ciliary dysfunction results in diseases (ciliopathies) that can affect all organs. Thanks to its complex ciliary structure, the unicellular and ciliated microorganism, Paramecium, constitutes a model of choice not only to study the structure, assembly and function of cilia but also to validate the specific role of mutations of genes linked to the ciliopathies.
    DOI:  https://doi.org/10.1051/medsci/2021087
  22. Cells. 2021 Jun 11. pii: 1463. [Epub ahead of print]10(6):
    Effect of Rho-Associated Kinase Inhibitor and Mesenchymal Stem Cell-Derived Conditioned Medium on Corneal Endothelial Cell Senescence and Proliferation.
    Boyoung Jung, Hun Lee, Sumi Kim, Hungwon Tchah, Changmo Hwang.
      This study aims to obtain sufficient corneal endothelial cells for regenerative application. We examined the combinatory effects of Rho-associated kinase (ROCK) inhibitor Y-27632 and mesenchymal stem cell-derived conditioned medium (MSC-CM) on the proliferation and senescence of rabbit corneal endothelial cells (rCECs). rCECs were cultured in a control medium, a control medium mixed with either Y-27632 or MSC-CM, and a combinatory medium containing Y-27632 and MSC-CM. Cells were analyzed for morphology, cell size, nuclei/cytoplasmic ratio, proliferation capacity and gene expression. rCECs cultured in a combinatory culture medium showed a higher passage number, cell proliferation, and low senescence. rCECs on collagen type I film showed high expression of tight junction. The cell proliferation marker Ki-67 was positively stained either in Y-27632 or MSC-CM-containing media. Genes related to cell proliferation resulted in negligible changes in MKI67, CIP2A, and PCNA in the combinatory medium, suggesting proliferative capacity was maintained. In contrast, all of these genes were significantly downregulated in the other groups. Senescence marker β-galactosidase-positive cells significantly decreased in either MSC-CM and/or Y-27632 mixed media. Senescence-related genes downregulated LMNB1 and MAP2K6, and upregulated MMP2. Cell cycle checkpoint genes such as CDC25C, CDCA2, and CIP2A did not vary in the combinatory medium but were significantly downregulated in either ROCK inhibitor or MSC-CM alone. These results imply the synergistic effect of combinatory culture medium on corneal endothelial cell proliferation and high cell number. This study supports high potential for translation to the development of human corneal endothelial tissue regeneration.
    Keywords:  ROCK inhibitor; Y-27632; mesenchymal stem cell-derived conditioned medium; rabbit corneal endothelial cells; senescence
    DOI:  https://doi.org/10.3390/cells10061463
  23. Front Cell Dev Biol. 2021 ;9 647391
    Centriolar Protein C2cd3 Is Required for Craniofacial Development.
    Ching-Fang Chang, Kari M Brown, Yanfen Yang, Samantha A Brugmann.
      The primary cilium is a ubiquitous, microtubule-based cellular organelle. Primary cilia dysfunction results in a group of disorders termed ciliopathies. C2 domain containing 3 centriole elongation regulator (C2cd3), encodes a centriolar protein essential for ciliogenesis. Mutations in human C2CD3 are associated with the human ciliopathy Oral-Facial-Digital syndrome type 14 (OFD14). In order to better understand the etiology of ciliopathies including OFD14, we generated numerous murine models targeting C2cd3. Initial analysis revealed several tissue-specific isoforms of C2cd3, and while the loss of C2cd3 has previously been reported to result in exencephaly, tight mesencephalic flexure, pericardial edema, abnormal heart looping and a twisted body axis, further analysis revealed that genetic background may also contribute to phenotypic variation. Additional analyses of a conditional allelic series targeting C-terminal PKC-C2 domains or the N-terminal C2CD3N-C2 domain of C2cd3 revealed a variable degree of phenotypic severity, suggesting that while the N-terminal C2CD3N-C2 domain was critical for early embryonic development as a whole, there was also a craniofacial specific role for the C2CD3N-C2 domains. Together, through generation of novel models and evaluation of C2cd3 expression, these data provide valuable insight into mechanisms of pathology for craniofacial ciliopathies that can be further explored in the future.
    Keywords:  C2cd3; ciliopathies; craniofacial development; neural crest; primary cilia
    DOI:  https://doi.org/10.3389/fcell.2021.647391
  24. J Neurosci. 2021 Jun 30. pii: JN-RM-3005-20. [Epub ahead of print]
    Multifaceted functions of Rab23 on primary cilium- and Hedgehog signaling-mediated cerebellar granule cell proliferation.
    Chh Hor, Jcw Lo, Als Cham, W Y Leong, Elk Goh.
      Sonic Hedgehog (Shh) signaling from the primary cilium drives cerebellar granule cell precursor (GCP) proliferation. Mutations of hedgehog (Hh) pathway repressors commonly cause medulloblastoma, the most prevalent and malignant childhood brain tumor that arises from aberrant GCP proliferation. We demonstrate that Nestin Cre-driven conditional knockout of a Shh pathway repressor-Rab23 in the mouse brain of both genders caused mis-patterning of cerebellar folia and elevated GCP proliferation during early development, but with no prevalent occurrence of medulloblastoma at adult stage. Strikingly, Rab23-depleted GCPs exhibited up-regulated basal level of Shh pathway activities despite showing an abnormal ciliogenesis of primary cilia. In line with the compromised ciliation, Rab23-depleted GCPs were desensitized against Hh pathway activity stimulations by Shh ligand and Smoothened (Smo) agonist-SAG, and exhibited attenuated stimulation of Smo-localization on the primary cilium in response to SAG. These results implicate multidimensional actions of Rab23 on Hh signaling cascade. Rab23 represses the basal level of Shh signaling, while facilitating primary cilium-dependent extrinsic Shh signaling activation. Collectively, our findings unravel instrumental roles of Rab23 in GCP proliferation and ciliogenesis. Furthermore, Rab23's potentiation of Shh signaling pathway through the primary cilium and Smo suggests a potential new therapeutic strategy for Smo/primary cilium-driven medulloblastoma.SIGNIFICANCE STATEMENT:Primary cilium and Sonic hedgehog (Shh) signaling are known to regulate GCP proliferation. Aberrant overactivation of Shh signaling pathway ectopically increases GCP proliferation and causes malignant childhood tumor called medulloblastoma. However, the genetic and molecular regulatory cascade of GCP tumorigenesis remains incompletely understood. Our finding uncovers Rab23 as a novel regulator of Hh signaling pathway activity and cell proliferation in GCP. Intriguingly, we demonstrated that Rab23 confers dual functions in regulating Shh signaling; it potentiates primary cilium and Shh/Smo-dependent signaling activation, while antagonizes basal level Hh activity. Our data present a previously underappreciated aspect of Rab23 in mediating extrinsic Shh signaling upstream of Smo. This study sheds new light on the mechanistic insights underpinning Shh signaling-mediated GCP proliferation and tumorigenesis.
    DOI:  https://doi.org/10.1523/JNEUROSCI.3005-20.2021
  25. Cells. 2021 Jun 11. pii: 1466. [Epub ahead of print]10(6):
    Etoposide Triggers Cellular Senescence by Inducing Multiple Centrosomes and Primary Cilia in Adrenocortical Tumor Cells.
    Yen-Ni Teng, Huei-Cih Chang, Yu-Ying Chao, Hui-Ling Cheng, Wei-Chih Lien, Chia-Yih Wang.
      Etoposide (ETO) has been used in treating adrenocortical tumor (ACT) cells. Our previous study showed that ETO inhibits ACT cell growth. In the present study, we show that ETO treatment at IC50 (10 μM) inhibited ACT cell growth by inducing cellular senescence rather than apoptosis. Several markers of cellular senescence, including enlarged nuclei, activated senescence-associated β-galactosidase activity, elevated levels of p53 and p21, and down-regulation of Lamin B1, were observed. We further found that ETO induced multiple centrosomes. The inhibition of multiple centrosomes accomplished by treating cells with either roscovitine or centrinone or through the overexpression of NR5A1/SF-1 alleviated ETO-induced senescence, suggesting that ETO triggered senescence via multiple centrosomes. Primary cilia also played a role in ETO-induced senescence. In the mechanism, DNA-PK-Chk2 signaling was activated by ETO treatment; inhibition of this signaling cascade alleviated multiple ETO-induced centrosomes and primary cilia followed by reducing cellular senescence. In addition to DNA damage signaling, autophagy was also triggered by ETO treatment for centrosomal events and senescence. Importantly, the inactivation of DNA-PK-Chk2 signaling reduced ETO-triggered autophagy; however, the inhibition of autophagy did not affect DNA-PK-Chk2 activation. Thus, ETO activated the DNA-PK-Chk2 cascade to facilitate autophagy. The activated autophagy further induced multiple centrosomes and primary cilia followed by triggering senescence.
    Keywords:  Chk2; DNA-PK; autophagy; centrosome; etoposide; primary cilia; senescence
    DOI:  https://doi.org/10.3390/cells10061466
  26. Cells. 2021 Jun 16. pii: 1518. [Epub ahead of print]10(6):
    Trace Amine-Associated Receptor 1 Trafficking to Cilia of Thyroid Epithelial Cells.
    Maria Qatato, Vaishnavi Venugopalan, Alaa Al-Hashimi, Maren Rehders, Aaron D Valentine, Zeynep Hein, Uillred Dallto, Sebastian Springer, Klaudia Brix.
      Trace amine-associated receptor 1 (rodent Taar1/human TAAR1) is a G protein-coupled receptor that is mainly recognized for its functions in neuromodulation. Previous in vitro studies suggested that Taar1 may signal from intracellular compartments. However, we have shown Taar1 to localize apically and on ciliary extensions in rodent thyrocytes, suggesting that at least in the thyroid, Taar1 may signal from the cilia at the apical plasma membrane domain of thyrocytes in situ, where it is exposed to the content of the follicle lumen containing putative Taar1 ligands. This study was designed to explore mouse Taar1 (mTaar1) trafficking, heterologously expressed in human and rat thyroid cell lines in order to establish an in vitro system in which Taar1 signaling from the cell surface can be studied in future. The results showed that chimeric mTaar1-EGFP traffics to the apical cell surface and localizes particularly to spherical structures of polarized thyroid cells, procilia, and primary cilia upon serum-starvation. Moreover, mTaar1-EGFP appears to form high molecular mass forms, possibly homodimers and tetramers, in stably expressing human thyroid cell lines. However, only monomeric mTaar1-EGFP was cell surface biotinylated in polarized human thyrocytes. In polarized rat thyrocytes, mTaar1-EGFP is retained in the endoplasmic reticulum, while cilia were reached by mTaar1-EGFP transiently co-expressed in combination with an HA-tagged construct of the related mTaar5. We conclude that Taar1 trafficking to cilia depends on their integrity. The results further suggest that an in vitro cell model was established that recapitulates Taar1 trafficking in thyrocytes in situ, in principle, and will enable studying Taar1 signaling in future, thus extending our general understanding of its potential significance for thyroid autoregulation.
    Keywords:  G protein-coupled receptors; cilia; green fluorescent protein; thyroid auto-regulation; thyroid epithelial cells; trace amine-associated receptor 1; trafficking
    DOI:  https://doi.org/10.3390/cells10061518
  27. Cells. 2021 Jun 30. pii: 1648. [Epub ahead of print]10(7):
    ROCK Inhibition as Potential Target for Treatment of Pulmonary Hypertension.
    Tadeu L Montagnoli, Jaqueline S da Silva, Susumu Z Sudo, Aimeé D Santos, Gabriel F Gomide, Mauro P L de Sá, Gisele Zapata-Sudo.
      Pulmonary hypertension (PH) is a cardiovascular disease caused by extensive vascular remodeling in the lungs, which ultimately leads to death in consequence of right ventricle (RV) failure. While current drugs for PH therapy address the sustained vasoconstriction, no agent effectively targets vascular cell proliferation and tissue inflammation. Rho-associated protein kinases (ROCKs) emerged in the last few decades as promising targets for PH therapy, since ROCK inhibitors demonstrated significant anti-remodeling and anti-inflammatory effects. In this review, current aspects of ROCK inhibition therapy are discussed in relation to the treatment of PH and RV dysfunction, from cell biology to preclinical and clinical studies.
    Keywords:  ROCK; pulmonary hypertension; right ventricle dysfunction
    DOI:  https://doi.org/10.3390/cells10071648
  28. Diagnostics (Basel). 2021 Jun 09. pii: 1063. [Epub ahead of print]11(6):
    UA-Zero as a Uranyl Acetate Replacement When Diagnosing Primary Ciliary Dyskinesia by Transmission Electron Microscopy.
    Andreia Lucia Pinto, Ranjit Kaur Rai, Amelia Shoemark, Claire Hogg, Thomas Burgoyne.
      Primary ciliary dyskinesia (PCD) is a disorder affecting motile cilia. An early accurate diagnosis helps prevent lung damage and preserve lung function. To make a diagnostic assessment, one of the commonly used methods that allows for the examination of ciliary ultrastructure is transmission electron microscopy (TEM). This allows for a quantitative assessment of ciliary components to identify defects associated with PCD. Heavy metal staining is required to provide a contrast when imaging cilia in the TEM. One of the most commonly used stains is uranyl acetate (UA). UA can be applied to cellular material before embedding (en bloc), or to ultrathin sections of embedded samples (grid staining). UA is radioactive and, due to growing safety concerns and restrictions by government bodies, universities and hospitals, it is essential to find a suitable alternative. We show UA-zero (UAZ), when used en bloc, provides a high contrast and is a suitable replacement for UA. PCD diagnostic experts, having reviewed ciliary cross-sections stained with UAZ en bloc, are confident that the staining and PCD defects are readily detectable similar to samples that have been stained with UA.
    Keywords:  diagnosis; electron microscopy; primary ciliary dyskinesia; uranyl acetate
    DOI:  https://doi.org/10.3390/diagnostics11061063
  29. Cells. 2021 Jun 24. pii: 1592. [Epub ahead of print]10(7):
    Regulation of the Actin Cytoskeleton via Rho GTPase Signalling in Dictyostelium and Mammalian Cells: A Parallel Slalom.
    Vedrana Filić, Lucija Mijanović, Darija Putar, Antea Talajić, Helena Ćetković, Igor Weber.
      Both Dictyostelium amoebae and mammalian cells are endowed with an elaborate actin cytoskeleton that enables them to perform a multitude of tasks essential for survival. Although these organisms diverged more than a billion years ago, their cells share the capability of chemotactic migration, large-scale endocytosis, binary division effected by actomyosin contraction, and various types of adhesions to other cells and to the extracellular environment. The composition and dynamics of the transient actin-based structures that are engaged in these processes are also astonishingly similar in these evolutionary distant organisms. The question arises whether this remarkable resemblance in the cellular motility hardware is accompanied by a similar correspondence in matching software, the signalling networks that govern the assembly of the actin cytoskeleton. Small GTPases from the Rho family play pivotal roles in the control of the actin cytoskeleton dynamics. Indicatively, Dictyostelium matches mammals in the number of these proteins. We give an overview of the Rho signalling pathways that regulate the actin dynamics in Dictyostelium and compare them with similar signalling networks in mammals. We also provide a phylogeny of Rho GTPases in Amoebozoa, which shows a variability of the Rho inventories across different clades found also in Metazoa.
    Keywords:  Amoebozoa; Rac; Rho; cell migration; cell motility; convergent evolution
    DOI:  https://doi.org/10.3390/cells10071592
  30. Lab Chip. 2021 Jun 28.
    Microfluidic chip grafted with integrin tension sensors for evaluating the effects of flowing shear stress and ROCK inhibitor on platelets.
    Subin Mao, Anwesha Sarkar, Yongliang Wang, Chao Song, Dana LeVine, Xuefeng Wang, Long Que.
      Integrins are key players in platelet adhesion and aggregation. Integrin molecular tensions, the forces transmitted by integrin molecules, are regulated by both mechanical and biochemical cues, and the outside-in and inside-out signaling has been extensively studied. While the mechanical properties of platelets at static status have been studied by atomic force microscopy, traction force microscopy and tension sensors, the biomechanical properties of flowing platelets remain elusive. Herein, we report microfluidic chips grafted with integrin tension sensors for microfluidic-force mapping in platelets. Specifically, the process of integrin αIIbβ3 mediating tension transmission and platelet adhesion under low flow rates has been obtained, and the process of platelet clustering at post-stenotic regions has been demonstrated. We found that flowing shear force can postpone the integrin-mediated tension transmission and platelet adhesion. We further evaluated the effect of Y-27632, a ROCK inhibitor that has been proven to reduce integrin-mediated platelet adhesion, at a series of concentrations and demonstrated that microfluidic chips with integrin tension sensors are sensitive to the concentration-dependent effects of Y-27632. Given their low cost and scalable throughput, these chips are ideal technical platforms for biological studies of platelets at flowing status and for platelet inhibitor or potential antiplatelet drug screening.
    DOI:  https://doi.org/10.1039/d1lc00259g
  31. Prog Mol Biol Transl Sci. 2021 ;pii: S1877-1173(21)00038-7. [Epub ahead of print]182 353-401
    Gene and epigenetic editing in the treatment of primary ciliopathies.
    Elisa Molinari, John A Sayer.
      Primary ciliopathies are inherited human disorders that arise from mutations in ciliary genes. They represent a spectrum of severe, incurable phenotypes, differentially involving several organs, including the kidney and the eye. The development of gene-based therapies is opening up new avenues for the treatment of ciliopathies. Particularly attractive is the possibility of correcting in situ the causative genetic mutation, or pathological epigenetic changes, through the use of gene editing tools. Due to their versatility and efficacy, CRISPR/Cas-based systems represent the most promising gene editing toolkit for clinical applications. However, delivery and specificity issues have so far held back the translatability of CRISPR/Cas-based therapies into clinical practice, especially where systemic administration is required. The eye, with its characteristics of high accessibility and compartmentalization, represents an ideal target for in situ gene correction. Indeed, studies for the evaluation of a CRISPR/Cas-based therapy for in vivo gene correction to treat a retinal ciliopathy have reached the clinical stage. Further technological advances may be required for the development of in vivo CRISPR-based treatments for the kidney. We discuss here the possibilities and the challenges associated to the implementation of CRISPR/Cas-based therapies for the treatment of primary ciliopathies with renal and retinal phenotypes.
    Keywords:  CRISPR; Ciliopathies; Cystic kidney disease; Epigenetic; Gene editing; Gene therapy; Retinal degeneration
    DOI:  https://doi.org/10.1016/bs.pmbts.2021.01.027
  32. Int J Mol Sci. 2021 Jun 03. pii: 6040. [Epub ahead of print]22(11):
    MRTF: Basic Biology and Role in Kidney Disease.
    Maria Zena Miranda, Zsuzsanna Lichner, Katalin Szászi, András Kapus.
      A lesser known but crucially important downstream effect of Rho family GTPases is the regulation of gene expression. This major role is mediated via the cytoskeleton, the organization of which dictates the nucleocytoplasmic shuttling of a set of transcription factors. Central among these is myocardin-related transcription factor (MRTF), which upon actin polymerization translocates to the nucleus and binds to its cognate partner, serum response factor (SRF). The MRTF/SRF complex then drives a large cohort of genes involved in cytoskeleton remodeling, contractility, extracellular matrix organization and many other processes. Accordingly, MRTF, activated by a variety of mechanical and chemical stimuli, affects a plethora of functions with physiological and pathological relevance. These include cell motility, development, metabolism and thus metastasis formation, inflammatory responses and-predominantly-organ fibrosis. The aim of this review is twofold: to provide an up-to-date summary about the basic biology and regulation of this versatile transcriptional coactivator; and to highlight its principal involvement in the pathobiology of kidney disease. Acting through both direct transcriptional and epigenetic mechanisms, MRTF plays a key (yet not fully appreciated) role in the induction of a profibrotic epithelial phenotype (PEP) as well as in fibroblast-myofibroblast transition, prime pathomechanisms in chronic kidney disease and renal fibrosis.
    Keywords:  Rho GTPases; actin cytoskeleton; gene expression; kidney fibrosis; myofibroblast; nucleocytoplasmic shuttling; profibrotic epithelial phenotype; transcription factors
    DOI:  https://doi.org/10.3390/ijms22116040
  33. Int J Mol Sci. 2021 Jun 29. pii: 7036. [Epub ahead of print]22(13):
    Exposure to 16 Hz Pulsed Electromagnetic Fields Protect the Structural Integrity of Primary Cilia and Associated TGF-β Signaling in Osteoprogenitor Cells Harmed by Cigarette Smoke.
    Yangmengfan Chen, Romina H Aspera-Werz, Maximilian M Menger, Karsten Falldorf, Michael Ronniger, Christina Stacke, Tina Histing, Andreas K Nussler, Sabrina Ehnert.
      Cigarette smoking (CS) is one of the main factors related to avoidable diseases and death across the world. Cigarette smoke consists of numerous toxic compounds that contribute to the development of osteoporosis and fracture nonunion. Exposure to pulsed electromagnetic fields (PEMF) was proven to be a safe and effective therapy to support bone fracture healing. The aims of this study were to investigate if extremely low frequency (ELF-) PEMFs may be beneficial to treat CS-related bone disease, and which effect the duration of the exposure has. In this study, immortalized human mesenchymal stem cells (SCP-1 cells) impaired by 5% cigarette smoke extract (CSE) were exposed to ELF-PEMFs (16 Hz) with daily exposure ranging from 7 min to 90 min. Cell viability, adhesion, and spreading were evaluated by Sulforhodamine B, Calcein-AM staining, and Phalloidin-TRITC/Hoechst 33342 staining. A migration assay kit was used to determine cell migration. Changes in TGF-β signaling were evaluated with an adenoviral Smad2/3 reporter assay, RT-PCR, and Western blot. The structure and distribution of primary cilia were analyzed with immunofluorescent staining. Our data indicate that 30 min daily exposure to a specific ELF-PEMF most effectively promoted cell viability, enhanced cell adhesion and spreading, accelerated migration, and protected TGF-β signaling from CSE-induced harm. In summary, the current results provide evidence that ELF-PEMF can be used to support early bone healing in patients who smoke.
    Keywords:  Extremely low frequency pulsed electromagnetic fields (ELF-PEMFs); TGF-β signaling; bone healing; cigarette smoke extract; mesenchymal stem cells; primary cilium
    DOI:  https://doi.org/10.3390/ijms22137036
  34. Development. 2021 Jul 01. pii: dev.199685. [Epub ahead of print]
    Early perturbation of Wnt signaling reveals patterning and invagination-evagination control points in molar tooth development.
    Rebecca Kim, Tingsheng Yu, Jingjing Li, Jan Prochazka, Amnon Sharir, Jeremy B A Green, Ophir D Klein.
      Tooth formation requires complex signaling interactions both within the oral epithelium and between the epithelium and the underlying mesenchyme. Previous studies of the Wnt/β-catenin pathway have shown that tooth formation is partly inhibited in loss-of-function mutants, and gain-of-function mutants have perturbed tooth morphology. However, the stage at which Wnt signaling is first important in tooth formation remains unclear. Here, using an Fgf8-promoter-driven and therefore early deletion of β-catenin in molar epithelium, we found that loss of Wnt/β-catenin signaling completely deletes the molar tooth, demonstrating that this pathway is central to the earliest stages of tooth formation. Early expression of a dominant-active β-catenin protein also perturbs tooth formation, producing a large domed evagination at early stages and supernumerary teeth later on. The early evaginations are associated with premature mesenchymal condensation marker, and are reduced by inhibition of condensation-associated collagen synthesis. We propose that invagination versus evagination morphogenesis is regulated by the relative timing of epithelial versus mesenchymal cell convergence regulated by canonical Wnt signaling. Together, these studies reveal new aspects of Wnt/β-catenin signaling in tooth formation and epithelial morphogenesis more broadly.
    Keywords:  Epithelial invagination; Morphogenesis; Tooth development; Wnt signaling
    DOI:  https://doi.org/10.1242/dev.199685
  35. Int J Mol Sci. 2021 Jun 17. pii: 6489. [Epub ahead of print]22(12):
    New Structural Perspectives in G Protein-Coupled Receptor-Mediated Src Family Kinase Activation.
    Sandra Berndt, Ines Liebscher.
      Src family kinases (SFKs) are key regulators of cell proliferation, differentiation, and survival. The expression of these non-receptor tyrosine kinases is strongly correlated with cancer development and tumor progression. Thus, this family of proteins serves as an attractive drug target. The activation of SFKs can occur via multiple signaling pathways, yet many of them are poorly understood. Here, we summarize the current knowledge on G protein-coupled receptor (GPCR)-mediated regulation of SFKs, which is of considerable interest because GPCRs are among the most widely used pharmaceutical targets. This type of activation can occur through a direct interaction between the two proteins or be allosterically regulated by arrestins and G proteins. We postulate that a rearrangement of binding motifs within the active conformation of arrestin-3 mediates Src regulation by comparison of available crystal structures. Therefore, we hypothesize a potentially different activation mechanism compared to arrestin-2. Furthermore, we discuss the probable direct regulation of SFK by GPCRs and investigate the intracellular domains of exemplary GPCRs with conserved polyproline binding motifs that might serve as scaffolding domains to allow such a direct interaction. Large intracellular domains in GPCRs are often understudied and, in general, not much is known of their contribution to different signaling pathways. The suggested direct interaction between a GPCR and a SFK could allow for a potential immediate allosteric regulation of SFKs by GPCRs and thereby unravel a novel mechanism of SFK signaling. This overview will help to identify new GPCR-SFK interactions, which could serve to explain biological functions or be used to modulate downstream effectors.
    Keywords:  G protein-coupled receptors; G proteins; GPCR; SFK; SH3 domains; Src kinases; allosteric regulation; arrestin; biased signaling; kinase activation; non-receptor tyrosine kinases; polyproline motifs; signaling
    DOI:  https://doi.org/10.3390/ijms22126489
  36. Arch Iran Med. 2021 May 01. 24(5): 364-373
    CEP104 and CEP290; Genes with Ciliary Functions Cause Intellectual Disability in Multiple Families.
    Shahrouz Khoshbakht, Maryam Beheshtian, Zohreh Fattahi, Niloofar Bazazzadegan, Elham Parsimehr, Mahsa Fadaee, Raheleh Vazehan, Mehrshid Faraji Zonooz, Ayda Abolhassani, Mina Makvand, Ariana Kariminejad, Arzu Celik, Kimia Kahrizi, Hossein Najmabadi.
       BACKGROUND: Neurodevelopmental and intellectual impairments are extremely heterogeneous disorders caused by a diverse variety of genes involved in different molecular pathways and networks. Genetic alterations in cilia, highly-conserved organelles with sensorineural and signal transduction roles can compromise their proper functions and lead to so-called "ciliopathies" featuring intellectual disability (ID) or neurodevelopmental disorders as frequent clinical manifestations. Here, we report several Iranian families affected with ID and other ciliopathy-associated features carrying known and novel variants in two ciliary genes; CEP104 and CEP290.
    METHODS: Whole exome and Targeted exome sequencing were carried out on affected individuals. Lymphoblastoid cell lines (LCLs) derived from the members of affected families were established for two families carrying CEP104 mutations. RNA and protein expression studies were carried out on these cells using qPCR and Western blot, respectively.
    RESULTS: A novel homozygous variant; NM_025114.3:c.7341_7344dupACTT p.(Ser2449Thrfs*8) and four previously reported homozygous variants; NM_025114.3:c.322C>T p.(Arg108*), NM_025114.3:c.4393C>T p.(Arg1465*), NM_025114.3:c.5668G>T p.(Gly1890*) and NM_025114.3:c.1666dupA p.(Ile556Asnfs*20) were identified in CEP290. In two other families, two novel homozygous variants; NM_014704:c.2356_2357insTT p.(Cys786Phefs*11) and NM_014704:c.1901_1902insT p.(Leu634Phefs*33) were identified in CEP104, another ciliary gene. qPCR and Western blot analyses showed significantly lower levels of CEP104 transcripts and protein in patients compared to heterozygous or normal family members.
    CONCLUSION: We emphasize on the clinical variability and pleiotropic phenotypes due to variants of these genes. In conclusion, our findings support the pivotal role of these genes resulting in cognitive and neurodevelopmental features.
    Keywords:  CEP104; CEP290; Ciliopathies; Intellectual disability; Neurodevelopmental disorders
    DOI:  https://doi.org/10.34172/aim.2021.53
  37. Cancers (Basel). 2021 Jun 11. pii: 2920. [Epub ahead of print]13(12):
    MITF Promotes Cell Growth, Migration and Invasion in Clear Cell Renal Cell Carcinoma by Activating the RhoA/YAP Signal Pathway.
    Nayoung Kim, Solbi Kim, Myung-Won Lee, Heung-Jin Jeon, Hyewon Ryu, Jin-Man Kim, Hyo-Jin Lee.
      Microphthalmia-associated transcription factor (MITF) is a basic helix-loop-helix leucine zipper transcription factor involved in the lineage-specific regulation of melanocytes, osteoclasts and mast cells. MITF is also involved in the progression of melanomas and other carcinomas, including the liver, pancreas and lung. However, the role of MITF in clear cell renal cell carcinoma (ccRCC) is largely unknown. This study investigates the functional role of MITF in cancer and the molecular mechanism underlying disease progression in ccRCC. MITF knockdown inhibited cell proliferation and shifted the cell cycle in ccRCC cells. In addition, MITF knockdown reduced wound healing, cell migration and invasion compared with the controls. Conversely, MITF overexpression in SN12C and SNU482 cells increased cell migration and invasion. Overexpression of MITF activated the RhoA/YAP signaling pathway, which regulates cell proliferation and invasion, and increased YAP signaling promoted cell cycle-related protein expression. Additionally, tumor formation was impaired by MITF knockdown and enhanced by MITF overexpression in vivo. In summary, MITF expression was associated with aggressive tumor behavior, and increased the migratory and invasive capabilities of ccRCC cells. These effects were reversed by MITF suppression. These results suggest that MITF is a potential therapeutic target for the treatment of ccRCC.
    Keywords:  MITF; RhoA; YAP; cell cycle; invasion; migration; proliferation; renal cell carcinoma
    DOI:  https://doi.org/10.3390/cancers13122920
  38. Diagnostics (Basel). 2021 Jun 14. pii: 1088. [Epub ahead of print]11(6):
    Evaluation of a Clinical Index as a Predictive Tool for Primary Ciliary Dyskinesia.
    Vendula Martinů, Lucie Bořek-Dohalská, Žofia Varényiová, Jiří Uhlík, Václav Čapek, Petr Pohunek, Václav Koucký.
       BACKGROUND: In primary ciliary dyskinesia (PCD) there is no single diagnostic test. Different predictive tools have been proposed to guide referral of high-risk patients for further diagnostic workup. We aimed to test clinical index (CI) on a large unselected cohort and compare its characteristics with other widely used tools-PICADAR and NA-CDCF.
    METHODS: CI, PICADAR, and NA-CDCF scores were calculated in 1401 patients with suspected PCD referred to our center. Their predictive characteristics were analyzed using receiver operating characteristics (ROC) curves and compared to each other. Nasal nitric oxide (nNO) was measured in 569 patients older than 3 years.
    RESULTS: PCD was diagnosed in 67 (4.8%) patients. CI, PICADAR, and NA-CDCF scores were higher in PCD than in nonPCD group (all p < 0.001). The area under the ROC curve (AUC) for CI was larger than for NA-CDCF (p = 0.005); AUCPICADAR and AUCNA-CDCF did not differ (p = 0.093). An overlap in signs and symptoms among tools was identified. PICADAR could not be assessed in 86 (6.1%) patients without chronic wet cough. For CI laterality or congenital heart defects assessment was not necessary. nNO further improved predictive power of all three tools.
    CONCLUSION: CI is a feasible predictive tool for PCD that may outperform PICADAR and NA-CFCD.
    Keywords:  North America criteria defined clinical features (NA-CDCF); clinical index (CI); nasal nitric oxide (nNO); predictive tools; primary ciliary dyskinesia (PCD); primary ciliary dyskinesia rule (PICADAR)
    DOI:  https://doi.org/10.3390/diagnostics11061088
  39. Front Cell Dev Biol. 2021 ;9 673986
    WIP, YAP/TAZ and Actin Connections Orchestrate Development and Transformation in the Central Nervous System.
    Inés M Antón, Francisco Wandosell.
      YAP (Yes-associated protein) and TAZ (transcriptional coactivator with PDZ-binding motif) are transcription co-regulators that make up the terminal components of the Hippo signaling pathway, which plays a role in organ size control and derived tissue homeostasis through regulation of the proliferation, differentiation and apoptosis of a wide variety of differentiated and stem cells. Hippo/YAP signaling contributes to normal development of the nervous system, as it participates in self-renewal of neural stem cells, proliferation of neural progenitor cells and differentiation, activation and myelination of glial cells. Not surprisingly, alterations in this pathway underlie the development of severe neurological diseases. In glioblastomas, YAP and TAZ levels directly correlate with the amount of the actin-binding molecule WIP (WASP interacting protein), which regulates stemness and invasiveness. In neurons, WIP modulates cytoskeleton dynamics through actin polymerization/depolymerization and acts as a negative regulator of neuritogenesis, dendrite branching and dendritic spine formation. Our working hypothesis is that WIP regulates the YAP/TAZ pools using a Hippo-independent pathway. Thus, in this review we will present some of the data that links WIP, YAP and TAZ, with a focus on their function in cells from the central and peripheral nervous systems. It is hoped that a better understanding of the mechanisms involved in brain and nervous development and the pathologies that arise due to their alteration will reveal novel therapeutic targets for neurologic diseases.
    Keywords:  Hippo pathway; axonogenesis; cytoskeleton; glioblastoma; neuritogenesis; nuclear actin
    DOI:  https://doi.org/10.3389/fcell.2021.673986
  40. Sci Rep. 2021 Jun 29. 11(1): 13484
    Concurrent YAP/TAZ and SMAD signaling mediate vocal fold fibrosis.
    Ryosuke Nakamura, Nao Hiwatashi, Renjie Bing, Carina P Doyle, Ryan C Branski.
      Vocal fold (VF) fibrosis is a major cause of intractable voice-related disability and reduced quality of life. Excision of fibrotic regions is suboptimal and associated with scar recurrence and/or further iatrogenic damage. Non-surgical interventions are limited, putatively related to limited insight regarding biochemical events underlying fibrosis, and downstream, the lack of therapeutic targets. YAP/TAZ integrates diverse cell signaling events and interacts with signaling pathways related to fibrosis, including the TGF-β/SMAD pathway. We investigated the expression of YAP/TAZ following vocal fold injury in vivo as well as the effects of TGF-β1 on YAP/TAZ activity in human vocal fold fibroblasts, fibroblast-myofibroblast transition, and TGF-β/SMAD signaling. Iatrogenic injury increased nuclear localization of YAP and TAZ in fibrotic rat vocal folds. In vitro, TGF-β1 activated YAP and TAZ in human VF fibroblasts, and inhibition of YAP/TAZ reversed TGF-β1-stimulated fibroplastic gene upregulation. Additionally, TGF-β1 induced localization of YAP and TAZ in close proximity to SMAD2/3, and nuclear accumulation of SMAD2/3 was inhibited by a YAP/TAZ inhibitor. Collectively, YAP and TAZ were synergistically activated with the TGF-β/SMAD pathway, and likely essential for the fibroplastic phenotypic shift in VF fibroblasts. Based on these data, YAP/TAZ may evolve as an attractive therapeutic target for VF fibrosis.
    DOI:  https://doi.org/10.1038/s41598-021-92871-z
  41. Int J Mol Sci. 2021 Jun 29. pii: 6992. [Epub ahead of print]22(13):
    PRP4 Promotes Skin Cancer by Inhibiting Production of Melanin, Blocking Influx of Extracellular Calcium, and Remodeling Cell Actin Cytoskeleton.
    Muhammad Bilal Ahmed, Salman Ul Islam, Young Sup Lee.
      Pre-mRNA processing factor 4B (PRP4) has previously been shown to induce epithelial-mesenchymal transition (EMT) and drug resistance in cancer cell lines. As melanin plays an important photoprotective role in the risk of sun-induced skin cancers, we have investigated whether PRP4 can induce drug resistance and regulate melanin biosynthesis in a murine melanoma (B16F10) cell line. Cells were incubated with a crucial melanogenesis stimulator, alpha-melanocyte-stimulating hormone, followed by transfection with PRP4. This resulted in the inhibition of the production of melanin via the downregulation of adenylyl cyclase-cyclic adenosine 3',5'-monophosphate (AC)-(cAMP)-tyrosinase synthesis signaling pathway. Inhibition of melanin production by PRP4 leads to the promotion of carcinogenesis and induced drug resistance in B16F10 cells. Additionally, PRP4 overexpression upregulated the expression of β-arrestin 1 and desensitized the extracellular calcium-sensing receptor (CaSR), which in turn, inhibited the influx of extracellular Ca2+ ions. The decreased influx of Ca2+ was confirmed by a decreased expression level of calmodulin. We have demonstrated that transient receptor potential cation channel subfamily C member 1 was involved in the influx of CaSR-induced Ca2+ via a decreasing level of its expression. Furthermore, PRP4 overexpression downregulated the expression of AC, decreased the synthesis of cAMP, and modulated the actin cytoskeleton by inhibiting the expression of Ras homolog family member A (RhoA). Our investigation suggests that PRP4 inhibits the production of melanin in B16F10 cells, blocks the influx of Ca2+ through desensitization of CaSR, and modulates the actin cytoskeleton through downregulating the AC-cAMP pathway; taken together, these observations collectively lead to the promotion of skin carcinogenesis.
    Keywords:  PRP4; actin cytoskeleton; cAMP; calcium-sensing receptor; drug resistance; melanocyte
    DOI:  https://doi.org/10.3390/ijms22136992
  42. Mol Biol Cell. 2021 Jun 30. mbcE21020060
    Centriole and Golgi microtubule nucleation are dispensable for the migration of human neutrophil-like cells.
    Lucas C Klemm, Ryan A Denu, Laurel E Hind, Briana L Rocha-Gregg, Mark E Burkard, Anna Huttenlocher.
      Neutrophils migrate in response to chemoattractants to mediate host defense. Chemoattractants drive rapid intracellular cytoskeletal rearrangements including the radiation of microtubules from the microtubule-organizing center (MTOC) towards the rear of polarized neutrophils. Microtubules regulate neutrophil polarity and motility, but little is known about the specific role of MTOCs. To characterize the role of MTOCs on neutrophil motility we depleted centrioles in a well-established neutrophil-like cell line. Surprisingly, both chemical and genetic centriole depletion increased neutrophil speed and chemotactic motility, suggesting an inhibitory role for centrioles during directed migration. We also found that depletion of both centrioles and GM130-mediated Golgi microtubule nucleation did not impair neutrophil directed migration. Taken together, our findings demonstrate an inhibitory role for centrioles and a resilient MTOC system in motile human neutrophil-like cells. [Media: see text] [Media: see text] [Media: see text].
    DOI:  https://doi.org/10.1091/mbc.E21-02-0060
  43. Mol Med Rep. 2021 Sep;pii: 626. [Epub ahead of print]24(3):
    [Erratum] Regulation and mechanism of YAP/TAZ in the mechanical microenvironment of stem cells (Review).
    Ying Li, Jinming Wang, Weiliang Zhong.
      Owing to an error that was made during the production stages of the above review article, what was actually Fig. 2 was inadvertently duplicated on p. 7 as Fig. 9. Fig. 9 as it should have appeared in the review is shown below. The Editor apologizes to the authors for this error, and regrets any inconvenience caused to the readership. [the original article was published in Molecular Medicine Reports 24: Article no. 506, 2021; DOI: 10.3892/mmr.2021.12145].
    Keywords:  mechanotransduction; microenvironment; signaling pathway; stem cells; yes‑associated protein/transcriptional coactivator with PDZ‑binding motif
    DOI:  https://doi.org/10.3892/mmr.2021.12265
  44. Nat Commun. 2021 Jul 01. 12(1): 4071
    Fluid flow-induced left-right asymmetric decay of Dand5 mRNA in the mouse embryo requires a Bicc1-Ccr4 RNA degradation complex.
    Katsura Minegishi, Benjamin Rothé, Kaoru R Komatsu, Hiroki Ono, Yayoi Ikawa, Hiromi Nishimura, Takanobu A Katoh, Eriko Kajikawa, Xiaorei Sai, Emi Miyashita, Katsuyoshi Takaoka, Kana Bando, Hiroshi Kiyonari, Tadashi Yamamoto, Hirohide Saito, Daniel B Constam, Hiroshi Hamada.
      Molecular left-right (L-R) asymmetry is established at the node of the mouse embryo as a result of the sensing of a leftward fluid flow by immotile cilia of perinodal crown cells and the consequent degradation of Dand5 mRNA on the left side. We here examined how the fluid flow induces Dand5 mRNA decay. We found that the first 200 nucleotides in the 3' untranslated region (3'-UTR) of Dand5 mRNA are necessary and sufficient for the left-sided decay and to mediate the response of a 3'-UTR reporter transgene to Ca2+, the cation channel Pkd2, the RNA-binding protein Bicc1 and their regulation by the flow direction. We show that Bicc1 preferentially recognizes GACR and YGAC sequences, which can explain the specific binding to a conserved GACGUGAC motif located in the proximal Dand5 3'-UTR. The Cnot3 component of the Ccr4-Not deadenylase complex interacts with Bicc1 and is also required for Dand5 mRNA decay at the node. These results suggest that Ca2+ currents induced by leftward fluid flow stimulate Bicc1 and Ccr4-Not to mediate Dand5 mRNA degradation specifically on the left side of the node.
    DOI:  https://doi.org/10.1038/s41467-021-24295-2
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