bims-bicyki 2021-05-02 papers

bims-bicyki

Biomed News

on Bicaudal-C1 and interactors in cystic kidney disease

Issue of 2021–05–02
thirty-two papers selected by
Céline Gagnieux, École Polytechnique Fédérale de Lausanne

IFT-A deficiency in juvenile mice impairs biliary development and exacerbates ADPKD liver disease.
Modeling Neoplastic Growth in Renal Cell Carcinoma and Polycystic Kidney Disease.
Simultaneous Use of ROCK Inhibitors and EP2 Agonists Induces Unexpected Effects on Adipogenesis and the Physical Properties of 3T3-L1 Preadipocytes.
Phosphorylation and Ubiquitylation Regulate Protein Trafficking, Signaling, and the Biogenesis of Primary Cilia.
Prognostic Value of Fibroblast Growth Factor 23 in Autosomal Dominant Polycystic Kidney Disease.
Per-Treatment Post Hoc Analysis of Clinical Trial Outcomes With Tolvaptan in ADPKD.
Factors predicting decline in renal function and kidney volume growth in autosomal dominant polycystic kidney disease: a prospective cohort study (Japanese Polycystic Kidney Disease registry: J-PKD).
Demographic, diagnostic and therapeutic characteristics of autosomal dominant polycystic kidney disease in Ghana.
HDAC6 Signaling at Primary Cilia Promotes Proliferation and Restricts Differentiation of Glioma Cells.
Multi-level regulation of β-catenin activity by SETD2 suppresses the transition from polycystic kidney disease to clear cell renal cell carcinoma.
Semi-Lethal Primary Ciliary Dyskinesia in Rats Lacking the Nme7 Gene.
Curcumin Inhibits Lysophosphatidic Acid Mediated MCP-1 Expression via Blocking ROCK Signalling.
Estrogen withdrawal alters cytoskeletal and primary ciliary dynamics resulting in increased Hedgehog and osteoclastogenic paracrine signalling in osteocytes.
The autophagy protein ATG16L1 cooperates with IFT20 and INPP5E to regulate the turnover of phosphoinositides at the primary cilium.
Polycystin-2 Is Required for Chondrocyte Mechanotransduction and Traffics to the Primary Cilium in Response to Mechanical Stimulation.
Loss of PKD1 and PKD2 share common effects on intracellular Ca2+ signaling.
Kinase-anchoring proteins in ciliary signal transduction.
Rho/ROCK-MYOCD in regulating airway smooth muscle growth and remodeling.
Hedgehog signaling and the primary cilium: implications for spatial and temporal constraints on signaling.
RhoA: a dubious molecule in cardiac pathophysiology.
AMPKα2 activation by an energy-independent signal ensures chromosomal stability during mitosis.
New Insights into Cellular Functions of Nuclear Actin.
Role of Rho GTPase Interacting Proteins in Subcellular Compartments of Podocytes.
Prostaglandin F2 Alpha Triggers the Disruption of Cell Adhesion with Cytokeratin and Vimentin in Bovine Luteal Theca Cells.
Severity and Duration of Acute Kidney Injury and Chronic Kidney Disease after Cardiac Surgery.
Hypoxia in chronic kidney disease: towards a paradigm shift?
Permissive effect of GSK3β on profibrogenic plasticity of renal tubular cells in progressive chronic kidney disease.
Myocardial Hypertrophy and Fibrosis Are Associated with Cardiomyocyte Beta-Catenin and TRPC6/Calcineurin/NFAT Signaling in Spontaneously Hypertensive Rats with 5/6 Nephrectomy.
Mechanically induced integrin ligation mediates intracellular calcium signaling with single pulsating cavitation bubbles.
Simulated Microgravity Inhibits the Proliferation of Chang Liver Cells by Attenuation of the Major Cell Cycle Regulators and Cytoskeletal Proteins.
Splice site m6A methylation prevents binding of U2AF35 to inhibit RNA splicing.
The Roles of Calmodulin and CaMKII in Cx36 Plasticity.

J Pathol. 2021 Apr 26.

IFT-A deficiency in juvenile mice impairs biliary development and exacerbates ADPKD liver disease.

Wei Wang, Tana S Pottorf, Henry H Wang, Ruochen Dong, Matthew A Kavanaugh, Joseph T Cornelius, Katie L Dennis, Udayan Apte, Michele T Pritchard, Madhulika Sharma, Pamela V Tran.

  Polycystic liver disease (PLD) is characterized by the growth of numerous biliary cysts and presents in patients with Autosomal Dominant Polycystic Kidney Disease (ADPKD), causing significant morbidity. Interestingly, deletion of intraflagellar transport-B (IFT-B) complex genes in adult mouse models of ADPKD attenuates severity of PKD and PLD. Here we examine the role of deletion of an IFT-A gene, Thm1, in PLD of juvenile and adult Pkd2 conditional knock-out mice. Perinatal deletion of Thm1 resulted in disorganized and expanded biliary regions, biliary fibrosis, increased serum bile acids, and a shortened primary cilium on CK19+ epithelial cells. In contrast, perinatal deletion of Pkd2 caused PLD, with multiple CK19+ epithelial cell-lined cysts, fibrosis, lengthened primary cilia, and increased Notch and ERK signaling. Perinatal deletion of Thm1 in Pkd2 conditional knock-out mice increased hepatomegaly, liver necrosis, as well as serum bilirubin and bile acid levels, indicating enhanced liver disease severity. In contrast to effects in the developing liver, deletion of Thm1 alone in adult mice did not cause a biliary phenotype. Combined deletion of Pkd2 and Thm1 caused variable hepatic cystogenesis at 4 months of age, but differences in hepatic cystogenesis between Pkd2- and Pkd2;Thm1 knock-out mice were not observed by 6-months of age. Similar to juvenile PLD, Notch and ERK signaling were increased in adult Pkd2 conditional knock-out cyst-lining epithelial cells. Taken together, Thm1 is required for biliary tract development, and proper biliary development restricts PLD severity. Unlike IFT-B genes, Thm1 does not markedly attenuate hepatic cystogenesis, suggesting differences in regulation of signaling and cystogenic processes in the liver by IFT-B and -A. Notably, increased Notch signaling in cyst-lining epithelial cells may indicate that aberrant activation of this pathway promotes hepatic cystogenesis, presenting as a novel potential therapeutic target. This article is protected by copyright. All rights reserved.

Keywords:  ADPKD; IFT-A; Notch signaling; Ttc21b; biliary ciliopathy; polycystic liver disease

DOI:  https://doi.org/10.1002/path.5685
Int J Mol Sci. 2021 Apr 10. pii: 3918. [Epub ahead of print]22(8):

Modeling Neoplastic Growth in Renal Cell Carcinoma and Polycystic Kidney Disease.

Cassandra Millet-Boureima, Stephanie He, Thi Bich Uyen Le, Chiara Gamberi.

  Renal cell carcinoma (RCC) and autosomal dominant polycystic kidney disease (ADPKD) share several characteristics, including neoplastic cell growth, kidney cysts, and limited therapeutics. As well, both exhibit impaired vasculature and compensatory VEGF activation of angiogenesis. The PI3K/AKT/mTOR and Ras/Raf/ERK pathways play important roles in regulating cystic and tumor cell proliferation and growth. Both RCC and ADPKD result in hypoxia, where HIF-α signaling is activated in response to oxygen deprivation. Primary cilia and altered cell metabolism may play a role in disease progression. Non-coding RNAs may regulate RCC carcinogenesis and ADPKD through their varied effects. Drosophila exhibits remarkable conservation of the pathways involved in RCC and ADPKD. Here, we review the progress towards understanding disease mechanisms, partially overlapping cellular and molecular dysfunctions in RCC and ADPKD and reflect on the potential for the agile Drosophila genetic model to accelerate discovery science, address unresolved mechanistic aspects of these diseases, and perform rapid pharmacological screens.

Keywords:  Drosophila; cilia; disease model; neoplasia; neovascularization; oxygen; pharmacology; polycystic kidney disease; renal cell carcinoma

DOI:  https://doi.org/10.3390/ijms22083918
Int J Mol Sci. 2021 Apr 28. pii: 4648. [Epub ahead of print]22(9):

Simultaneous Use of ROCK Inhibitors and EP2 Agonists Induces Unexpected Effects on Adipogenesis and the Physical Properties of 3T3-L1 Preadipocytes.

Yosuke Ida, Megumi Watanabe, Hiroshi Ohguro, Fumihito Hikage.

  To elucidate the additive effects of an EP2 agonist, omidenepag (OMD) or butaprost (Buta) on the Rho-associated coiled-coil-containing protein kinase (ROCK) inhibitor, ripasudil (Rip) on adipose tissue, two- or three-dimension (2D or 3D) cultures of 3T3-L1 cells were analyzed by lipid staining, the mRNA expression of adipogenesis-related genes, extracellular matrix (ECM) molecules including collagen (Col) -1, -4 and -6, and fibronectin (Fn), and the sizes and physical properties of 3D organoids, as measured by a micro-squeezer. The results indicate that adipogenesis induced (1) an enlargement of the 3D organoids; (2) a substantial enhancement in lipid staining as well as the expression of the Pparγ, Ap2 and Leptin genes; (3) a significant softening of the 3D organoids, the effects of which were all enhanced by Rip except for Pparγ expression; and (4) a significant downregulation in Col1 and Fn, and a significant upregulation in Col4, Col6, the effects of which were unchanged by Rip. When adding the EP2 agonist to Rip, (1) the sizes of the 3D organoids were reduced substantially; (2) lipid staining was increased (OMD), or decreased (Buta); (3) the stiffness of the 3D organoids was substantially increased in Buta; (4-1) the expression of Pparγ was suppressed (2D, OMD) or increased (2D, Buta), and the expressions of Ap2 were downregulated (2D, 3D) and Leptin was increased (2D) or decreased (3D), (4-2) all the expressions of four ECM molecules were upregulated in 2D (2D), and in 3D, the expression of Col1, Col4 was upregulated. The collective findings reported herein indicate that the addition of an EP2 agonist, OMD or Buta significantly but differently modulate the Rip-induced effects on adipogenesis and the physical properties of 2D and 3D cultured 3T3-L1 cells.

Keywords:  3-dimension tissue cultures; 3T3-L1 cell; EP2 agonist; ROCK inhibitor

DOI:  https://doi.org/10.3390/ijms22094648
Front Cell Dev Biol. 2021 ;9 664279

Phosphorylation and Ubiquitylation Regulate Protein Trafficking, Signaling, and the Biogenesis of Primary Cilia.

Elena A May, Tommy J Sroka, David U Mick.

  The primary cilium is a solitary, microtubule-based membrane protrusion extending from the surface of quiescent cells that senses the cellular environment and triggers specific cellular responses. The functions of primary cilia require not only numerous different components but also their regulated interplay. The cilium performs highly dynamic processes, such as cell cycle-dependent assembly and disassembly as well as delivery, modification, and removal of signaling components to perceive and process external signals. On a molecular level, these processes often rely on a stringent control of key modulatory proteins, of which the activity, localization, and stability are regulated by post-translational modifications (PTMs). While an increasing number of PTMs on ciliary components are being revealed, our knowledge on the identity of the modifying enzymes and their modulation is still limited. Here, we highlight recent findings on cilia-specific phosphorylation and ubiquitylation events. Shedding new light onto the molecular mechanisms that regulate the sensitive equilibrium required to maintain and remodel primary cilia functions, we discuss their implications for cilia biogenesis, protein trafficking, and cilia signaling processes.

Keywords:  Hedgehog signaling; cell signaling; ciliogenesis; phosphorylation; post-translational modification; primary cilia; ubiquitylation

DOI:  https://doi.org/10.3389/fcell.2021.664279
Kidney Int Rep. 2021 Apr;6(4): 953-961

Prognostic Value of Fibroblast Growth Factor 23 in Autosomal Dominant Polycystic Kidney Disease.

Mireille El Ters, Pengcheng Lu, Jonathan D Mahnken, Jason R Stubbs, Shiqin Zhang, Darren P Wallace, Jared J Grantham, Arlene B Chapman, Vicente E Torres, Peter C Harris, Kyongtae Ty Bae, Douglas P Landsittel, Frederic F Rahbari-Oskoui, Michal Mrug, William M Bennett, Alan S L Yu.

   Introduction: Autosomal dominant polycystic kidney disease (ADPKD) is characterized by progressive cyst growth and a loss of functioning renal mass, but a decline in glomerular filtration rate (GFR) and onset of end-stage renal disease (ESRD) occur late in the disease course. There is therefore a great need for early prognostic biomarkers in this disorder.
Methods: We measured baseline serum fibroblast growth factor 23 (FGF23) levels in 192 patients with ADPKD from the Consortium for Radiologic Imaging Studies of PKD (CRISP) cohort that were followed for a median of 13 years and tested the association between FGF23 levels and change over time in height-adjusted total kidney volume (htTKV), GFR, and time to the composite endpoints of ESRD, death, and doubling of serum creatinine.
Results: Patients in the highest quartile for baseline FGF23 level had a higher rate of increase in htTKV (0.95% per year, P = 0.0016), and faster rate of decline in GFR (difference of -1.03 ml/min/1.73 m2 per year, P = 0.005) compared with the lowest quartile, after adjusting for other covariates, including htTKV and genotype. The highest quartile of FGF23 was also associated with a substantial increase in risk for the composite endpoint of ESRD, death, or doubling of serum creatinine (hazard ratio [HR] of 2.45 in the fully adjusted model, P = 0.03).
Conclusion: FGF23 is a prognostic biomarker for disease progression and clinically important outcomes in ADPKD, and has additive value to established imaging and genetic biomarkers.

Keywords:  ADPKD; FGF23

DOI:  https://doi.org/10.1016/j.ekir.2021.01.004
Kidney Int Rep. 2021 Apr;6(4): 1032-1040

Per-Treatment Post Hoc Analysis of Clinical Trial Outcomes With Tolvaptan in ADPKD.

Andrew J Mallett, Ronald D Perrone, Gopala Rangan, Carmel Hawley, Ragada El-Damanawi, Thomas F Hiemstra, Carolina Townsend Arellano, Jennifer Lee, Vicente E Torres.

   Introduction: In pivotal trials of patients with autosomal dominant polycystic kidney disease at risk of rapid progression, tolvaptan slowed estimated glomerular filtration rate (eGFR) decline in early-to-moderate (TEMPO 3:4 [NCT00428948]) and moderate- to late-stage (REPRISE [NCT02160145]) chronic kidney disease (CKD). Discontinuation was less frequent in REPRISE (15.0%) than TEMPO 3:4 (23.0%), given that in REPRISE, only subjects who tolerated tolvaptan 60/30 mg daily initiated the double-blind phase. We evaluated whether the greater treatment effect in REPRISE was attributable to different completion rates.
Methods: We conducted post hoc analyses of TEMPO 3:4 and REPRISE completers, defined as subjects who took trial drug to the end of the treatment period in TEMPO 3:4 (3 years) or REPRISE (1 year). Efficacy (rate of change in eGFR for tolvaptan vs. placebo) was analyzed as in each trial. Subjects from TEMPO 3:4 and REPRISE were also matched by propensity score for age, gender, and baseline eGFR to explore potential additional determinants of treatment effect.
Results: The annualized tolvaptan treatment effect in TEMPO 3:4 completers (difference vs. placebo of 0.98 ml/min per 1.73 m2/y) and REPRISE completers (difference of 1.23) was similar to that of the respective total trial populations (TEMPO 3:4: 0.94; REPRISE: 1.27). The treatment effect of tolvaptan was also similar between matched subjects.
Conclusion: Greater treatment completion rate did not drive greater treatment effect in REPRISE. The more advanced CKD of REPRISE subjects may be more relevant. More rapid decline in kidney function in later-stage CKD enabled the effects of tolvaptan to be more easily discerned.

Keywords:  autosomal dominant polycystic kidney disease; chronic kidney disease; clinical trial; persistence; tolvaptan; treatment effect

DOI:  https://doi.org/10.1016/j.ekir.2021.01.014
Clin Exp Nephrol. 2021 Apr 29.

Factors predicting decline in renal function and kidney volume growth in autosomal dominant polycystic kidney disease: a prospective cohort study (Japanese Polycystic Kidney Disease registry: J-PKD).

Kiyotaka Uchiyama, Toshio Mochizuki, Yosuke Shimada, Saori Nishio, Hiroshi Kataoka, Michihiro Mitobe, Ken Tsuchiya, Kazushige Hanaoka, Yoshifumi Ubara, Tatsuya Suwabe, Akinari Sekine, Kikuo Nutahara, Kazuhiko Tsuruya, Eiji Ishimura, Shinya Nakatani, Tadashi Sofue, Satoshi Tanaka, Ichiei Narita, Shoichi Maruyama, Shigeo Horie, Satoru Muto.

   BACKGROUND: Factors affecting decline in renal function and cyst growth in patients with autosomal polycystic kidney disease (ADPKD) are not fully described, particularly in Japan.
METHODS: This was the first multi-facility, prospective, observational cohort study conducted in ADPKD patients at 14 centers in Japan. Patients in the J-PKD registry were assessed from December 2009 to June 2012 (follow-up until June 2017). Patients' data including estimated glomerular filtration rate (eGFR) and total kidney volume (TKV) were assessed initially and a maximum of five times annually. Contributing factors to eGFR decline and TKV growth were identified using multiple linear regression analysis.
RESULTS: Of the 340 patients in the J-PKD registry, data analysis was performed for 192 patients in whom serial changes for both eGFR and TKV were obtained. eGFR slope, eGFR change, and TKV change values were as follows: - 2.7 (- 4.2 to - 1.5) (ml/min/1.73 m2/year), - 5.0 (- 9.6 to - 2.3) (%/year), and 4.78 (0.86-8.22) (%/year), respectively. Lower high-density lipoprotein (HDL) cholesterol was an independent predictor of eGFR decline, using both eGFR slope and change (P = 0.04, P = 0.02, respectively), whereas lower hemoglobin and higher uric acid were significantly associated with greater eGFR change only (P = 0.02, P = 0.002, respectively). Younger age and higher fasting blood sugar were independent predictors of greater TKV change (P = 0.01, P = 0.02, respectively).
CONCLUSIONS: This real-world study in Japan identified risk factors for renal function decline in ADPKD patients. These included lower HDL cholesterol, lower hemoglobin and higher uric acid for eGFR decline, and youth and higher blood sugar levels for TKV growth.

Keywords:  Autosomal dominant polycystic kidney disease; High-density lipoprotein cholesterol; J-PKD; Renal function; Total kidney volume

DOI:  https://doi.org/10.1007/s10157-021-02068-x
BMC Nephrol. 2021 Apr 28. 22(1): 156

Demographic, diagnostic and therapeutic characteristics of autosomal dominant polycystic kidney disease in Ghana.

Perditer Okyere, Richard K D Ephraim, Isaac Okyere, Joseph Attakorah, Dorcas Serwaa, Grace Essuman, Albert Abaka-Yawson, Prince Adoba.

   BACKGROUND: Autosomal Dominant Polycystic Kidney Disease (ADPKD) is the commonest of the hereditary kidney diseases and mostly ensues in utero with signs delayed until after several decades. This study assessed the demographic, diagnostic (clinical and biochemical features) and therapeutic patterns among ADPKD patients who attended the nephrology unit of Komfo Anokye Teaching Hospital (KATH) from 2007 to 2018.
METHODS: This cross-sectional retrospective analysis of ADPKD patient records was conducted at the nephrology unit of KATH in October 2020. The records of 82 ADPKD was used for this study. Demographic, clinical, biochemical, ultrasonographic and therapeutic data was obtained, organized and analyzed with Statistical Package for the Social Sciences (SPSS).
RESULTS: ADPKD was most prevalent in people within the ages of 31-40 years (25.6 %), with a male (52.4 %) preponderance. The most common clinical features presented were flank pain (30.5 %) and bipedal swelling (18.3 %). Hypertension (42.7 %), urinary tract infections (UTIs) (19.5 %), and anemia (13.4 %) were the most common complications reported. Average level of HDL-c was higher in females (1.7) than in males (1.2) (p = 0.001). Hematuria (34 %) and proteinuria (66 %) were among the biochemical derangements presented. About 81.7 % had CKD at diagnosis with the majority in stages 1 (27.0 %), 3(23.2 %) and 5 (20.3 %). Poor corticomedullary differentiation was observed in 90.2 % of participants and increased echogenicity was observed in 89.0 % of the participants. Estimated GFR (eGFR) correlated positively with echotexture (r = 0.320, p = 0.005) and negatively with CMD (r= -0.303, p = 0.008). About 95.1 % of patients were on conservative therapy including: 73.2 %, 52.4 %, 22.0 %, 13.4 %, 8.5 % on Irebesartan/Lisinopril, Nifecard XL, Hydralazine, Methyldopa and Bisoprolol respectively for hypertension; 26.8 and 3.7 % on Gliclazide and Metformin respectively for Type 2 diabetes mellitus; 25.6 %, 24.4 and 18.3 % on CaCO3, fersolate and folic acid respectively as nutrient supplements with 4.9 % of participants on renal replacement therapy (RRT).
CONCLUSIONS: ADPKD occurs in people aged ≥ 31 years with a higher male preponderance. Clinical features include flank and abdominal pain, bipedal swelling, headache, amongst others. Uremia, hematuria, proteinuria, decreased eGFR, were the common biochemical derangements reported with higher severity detected in men. The therapeutic interventions mostly involved conservative therapy to manage symptoms and other comorbid conditions and rarely renal replacement therapy (RRT).

Keywords:  Autosomal dominant polycystic kidney disease; Biochemical derangements; Chronic kidney disease; Clinical features; Demographic characteristics

DOI:  https://doi.org/10.1186/s12882-021-02336-8
Cancers (Basel). 2021 Apr 01. pii: 1644. [Epub ahead of print]13(7):

HDAC6 Signaling at Primary Cilia Promotes Proliferation and Restricts Differentiation of Glioma Cells.

Ping Shi, Lan B Hoang-Minh, Jia Tian, Alice Cheng, Reemsha Basrai, Neil Kalaria, Joseph J Lebowitz, Habibeh Khoshbouei, Loic P Deleyrolle, Matthew R Sarkisian.

  Histone deacetylase 6 (HDAC6) is an emerging therapeutic target that is overexpressed in glioblastoma when compared to other HDACs. HDAC6 catalyzes the deacetylation of alpha-tubulin and mediates the disassembly of primary cilia, a process required for cell cycle progression. HDAC6 inhibition disrupts glioma proliferation, but whether this effect is dependent on tumor cell primary cilia is unknown. We found that HDAC6 inhibitors ACY-1215 (1215) and ACY-738 (738) inhibited the proliferation of multiple patient-derived and mouse glioma cells. While both inhibitors triggered rapid increases in acetylated alpha-tubulin (aaTub) in the cytosol and led to increased frequencies of primary cilia, they unexpectedly reduced the levels of aaTub in the cilia. To test whether the antiproliferative effects of HDAC6 inhibitors are dependent on tumor cell cilia, we generated patient-derived glioma lines devoid of cilia through depletion of ciliogenesis genes ARL13B or KIF3A. At low concentrations, 1215 or 738 did not decrease the proliferation of cilia-depleted cells. Moreover, the differentiation of glioma cells that was induced by HDAC6 inhibition did not occur after the inhibition of cilia formation. These data suggest HDAC6 signaling at primary cilia promotes the proliferation of glioma cells by restricting their ability to differentiate. Surprisingly, overexpressing HDAC6 did not reduce cilia length or the frequency of ciliated glioma cells, suggesting other factors are required to control HDAC6-mediated cilia disassembly in glioma cells. Collectively, our findings suggest that HDAC6 promotes the proliferation of glioma cells through primary cilia.

Keywords:  ARL13B; alpha-tubulin; glioblastoma; histone deacetylase 6; primary cilium

DOI:  https://doi.org/10.3390/cancers13071644
Cancer Res. 2021 Apr 28. pii: canres.3960.2020. [Epub ahead of print]

Multi-level regulation of β-catenin activity by SETD2 suppresses the transition from polycystic kidney disease to clear cell renal cell carcinoma.

Hanyu Rao, Xiaoxue Li, Min Liu, Jing Liu, Wenxin Feng, Huayuan Tang, Jin Xu, Wei-Qiang Gao, Li Li.

Patients with polycystic kidney disease (PKD) are at a high risk of developing renal cell carcinoma (RCC). However, little is known about genetic alterations or changes in signaling pathways during the transition from PKD to RCC. SET domain-containing 2 (SETD2) is a histone methyltransferase which catalyzes tri-methylation of H3K36 (H3K36me3) and has been identified as a tumor suppressor in clear cell renal cell carcinoma (ccRCC), but the underlying mechanism remains largely unexplored. Here we report that knockout of Setd2 in a c-MYC-driven PKD mouse model drove the transition to ccRCC. SETD2 inhibited β-catenin activity at transcriptional and post-transcriptional levels by competing with β-catenin for binding promoters of target genes and maintaining transcript levels of members of the β-catenin destruction complex. Thus, SETD2 deficiency enhanced the epithelial-mesenchymal transition and tumorigenesis through the hyperactivation of Wnt/β-catenin signaling. Our findings reveal previously unrecognized roles of SETD2-mediated competitive DNA binding and H3K36me3 modification in regulating Wnt/β-catenin signaling during the transition from PKD to ccRCC. The novel autochthonous mouse models of PKD and ccRCC will be useful for pre-clinical research into disease progression.

DOI: https://doi.org/10.1158/0008-5472.CAN-20-3960
Int J Mol Sci. 2021 Apr 07. pii: 3810. [Epub ahead of print]22(8):

Semi-Lethal Primary Ciliary Dyskinesia in Rats Lacking the Nme7 Gene.

Lucie Šedová, Ivana Buková, Pavla Bažantová, Silvia Petrezsélyová, Jan Prochazka, Elena Školníková, Dagmar Zudová, Josef Včelák, Pavol Makovický, Běla Bendlová, Ondřej Šeda, Radislav Sedlacek.

  NME7 (non-metastatic cells 7, nucleoside diphosphate kinase 7) is a member of a gene family with a profound effect on health/disease status. NME7 is an established member of the ciliome and contributes to the regulation of the microtubule-organizing center. We aimed to create a rat model to further investigate the phenotypic consequences of Nme7 gene deletion. The CRISPR/Cas9 nuclease system was used for the generation of Sprague Dawley Nme7 knock-out rats targeting the exon 4 of the Nme7 gene. We found the homozygous Nme7 gene deletion to be semi-lethal, as the majority of SDNme7-/- pups died prior to weaning. The most prominent phenotypes in surviving SDNme7-/- animals were hydrocephalus, situs inversus totalis, postnatal growth retardation, and sterility of both sexes. Thinning of the neocortex was histologically evident at 13.5 day of gestation, dilation of all ventricles was detected at birth, and an external sign of hydrocephalus, i.e., doming of the skull, was usually apparent at 2 weeks of age. Heterozygous SDNme7+/- rats developed normally; we did not detect any symptoms of primary ciliary dyskinesia. The transcriptomic profile of liver and lungs corroborated the histological findings, revealing defects in cell function and viability. In summary, the knock-out of the rat Nme7 gene resulted in a range of conditions consistent with the presentation of primary ciliary dyskinesia, supporting the previously implicated role of the centrosomally located Nme7 gene in ciliogenesis and control of ciliary transport.

Keywords:  Nme7; cilia; hydrocephalus; infertility; knock-out rat

DOI:  https://doi.org/10.3390/ijms22083810
Molecules. 2021 Apr 16. pii: 2320. [Epub ahead of print]26(8):

Curcumin Inhibits Lysophosphatidic Acid Mediated MCP-1 Expression via Blocking ROCK Signalling.

Ying Zhou, Peter J Little, Suowen Xu, Danielle Kamato.

  Curcumin is a natural compound that has been widely used as a food additive and medicine in Asian countries. Over several decades, diverse biological effects of curcumin have been elucidated, such as anti-inflammatory and anti-oxidative activities. Monocyte chemoattractant protein-1 (MCP-1) is a key inflammatory marker during the development of atherosclerosis, and curcumin blocks MCP-1 expression stimulated by various ligands. Hence, we studied the action of curcumin on lysophosphatidic acid (LPA) mediated MCP-1 expression and explored the specific underlying mechanisms. In human vascular smooth muscle cells, LPA induces Rho-associated protein kinase (ROCK) dependent transforming growth factor receptor (TGFBR1) transactivation, leading to glycosaminoglycan chain elongation. We found that LPA also signals via the TGFBR1 transactivation pathway to regulate MCP-1 expression. Curcumin blocks LPA mediated TGFBR1 transactivation and subsequent MCP-1 expression by blocking the ROCK signalling. In the vasculature, ROCK signalling regulates smooth muscle cell contraction, inflammatory cell recruitment, endothelial dysfunction and vascular remodelling. Therefore, curcumin as a ROCK signalling inhibitor has the potential to prevent atherogenesis via multiple ways.

Keywords:  Smad2; atherosclerosis; inflammation; monocyte chemoattractant protein-1; transforming growth factor receptor; vascular smooth muscle cells

DOI:  https://doi.org/10.3390/molecules26082320
Sci Rep. 2021 Apr 29. 11(1): 9272

Estrogen withdrawal alters cytoskeletal and primary ciliary dynamics resulting in increased Hedgehog and osteoclastogenic paracrine signalling in osteocytes.

Ivor P Geoghegan, Laoise M McNamara, David A Hoey.

Estrogen deficiency during post-menopausal osteoporosis leads to osteoclastogenesis and bone loss. Increased pro-osteoclastogenic signalling (RANKL/OPG) by osteocytes occurs following estrogen withdrawal (EW) and is associated with impaired focal adhesions (FAs) and a disrupted actin cytoskeleton. RANKL production is mediated by Hedgehog signalling in osteocytes, a signalling pathway associated with the primary cilium, and the ciliary structure is tightly coupled to the cytoskeleton. Therefore, the objective of this study was to investigate the role of the cilium and associated signalling in EW-mediated osteoclastogenic signalling in osteocytes. We report that EW leads to an elongation of the cilium and increase in Hedgehog and osteoclastogenic signalling. Significant trends were identified linking cilia elongation with reductions in cell area and % FA area/cell area, indicating that cilia elongation is associated with disruption of FAs and actin contractility. To verify this, we inhibited FA assembly via αvβ3 antagonism and inhibited actin contractility and demonstrated an elongated cilia and increased expression of Hh markers and Rankl expression. Therefore, our results suggest that the EW conditions associated with osteoporosis lead to a disorganisation of αvβ3 integrins and reduced actin contractility, which were associated with an elongation of the cilium, activation of the Hh pathway and osteoclastogenic paracrine signalling.

DOI: https://doi.org/10.1038/s41598-021-88633-6
Cell Rep. 2021 Apr 27. pii: S2211-1247(21)00361-2. [Epub ahead of print]35(4): 109045

The autophagy protein ATG16L1 cooperates with IFT20 and INPP5E to regulate the turnover of phosphoinositides at the primary cilium.

Asma Boukhalfa, Federica Roccio, Nicolas Dupont, Patrice Codogno, Etienne Morel.

  The primary cilium (PC) regulates signalization linked to external stress sensing. Previous works established a functional interplay between the PC and the autophagic machinery. When ciliogenesis is promoted by serum deprivation, the autophagy protein ATG16L1 and the ciliary protein IFT20 are co-transported to the PC. Here, we demonstrate that IFT20 and ATG16L1 are part of the same complex requiring the WD40 domain of ATG16L1 and a Y-E-F-I motif in IFT20. We show that ATG16L1-deficient cells exhibit aberrant ciliary structures, which accumulate PI4,5P2, whereas PI4P, a lipid normally concentrated in the PC, is absent. Finally, we demonstrate that INPP5E, a phosphoinositide-associated phosphatase responsible for PI4P generation, interacts with ATG16L1 and that a perturbation of the ATG16L1/IFT20 complex alters its trafficking to the PC. Altogether, our results reveal a function of ATG16L1 in ciliary lipid and protein trafficking, thus directly contributing to proper PC dynamics and functions.

Keywords:  ATG; IFT; INPP5E; PI4P; macroautophagy; phosphoinositides; primary cilium; trafficking

DOI:  https://doi.org/10.1016/j.celrep.2021.109045
Int J Mol Sci. 2021 Apr 21. pii: 4313. [Epub ahead of print]22(9):

Polycystin-2 Is Required for Chondrocyte Mechanotransduction and Traffics to the Primary Cilium in Response to Mechanical Stimulation.

Clare L Thompson, Megan McFie, J Paul Chapple, Philip Beales, Martin M Knight.

  Primary cilia and associated intraflagellar transport are essential for skeletal development, joint homeostasis, and the response to mechanical stimuli, although the mechanisms remain unclear. Polycystin-2 (PC2) is a member of the transient receptor potential polycystic (TRPP) family of cation channels, and together with Polycystin-1 (PC1), it has been implicated in cilia-mediated mechanotransduction in epithelial cells. The current study investigates the effect of mechanical stimulation on the localization of ciliary polycystins in chondrocytes and tests the hypothesis that they are required in chondrocyte mechanosignaling. Isolated chondrocytes were subjected to mechanical stimulation in the form of uniaxial cyclic tensile strain (CTS) in order to examine the effects on PC2 ciliary localization and matrix gene expression. In the absence of strain, PC2 localizes to the chondrocyte ciliary membrane and neither PC1 nor PC2 are required for ciliogenesis. Cartilage matrix gene expression (Acan, Col2a) is increased in response to 10% CTS. This response is inhibited by siRNA-mediated loss of PC1 or PC2 expression. PC2 ciliary localization requires PC1 and is increased in response to CTS. Increased PC2 cilia trafficking is dependent on the activation of transient receptor potential cation channel subfamily V member 4 (TRPV4) activation. Together, these findings demonstrate for the first time that polycystins are required for chondrocyte mechanotransduction and highlight the mechanosensitive cilia trafficking of PC2 as an important component of cilia-mediated mechanotransduction.

Keywords:  Polycystin; Polycystin-1; Polycystin-2; cartilage; chondrocyte; cilia; mechanotransduction; strain

DOI:  https://doi.org/10.3390/ijms22094313
Cell Calcium. 2021 Apr 24. pii: S0143-4160(21)00067-1. [Epub ahead of print]97 102413

Loss of PKD1 and PKD2 share common effects on intracellular Ca2+ signaling.

Ines Cabrita, Khaoula Talbi, Karl Kunzelmann, Rainer Schreiber.

  In polycystic kidney disease (PKD) multiple bilateral renal cysts gradually enlarge causing a decline in renal function. Transepithelial chloride secretion through cystic fibrosis transmembrane conductance regulator (CFTR) and TMEM16A (anoctamin 1) drive cyst enlargement. We demonstrated recently that a loss of PKD1 increases expression and function of TMEM16A in murine kidneys and in mouse M1 collecting duct cells. The data demonstrated that TMEM16A contributes essentially to cyst growth by upregulating intracellular Ca2+ signaling. Enhanced expression of TMEM16A and Ca2+ signaling increased both cell proliferation and fluid secretion, which suggested inhibition of TMEM16A as a novel therapy in ADPKD. About 15 % of all ADPKD cases are caused by mutations in PKD2. To analyze the effects of loss of function of PKD2 on Ca2+ signaling, we knocked-down Pkd2 in mouse primary renal epithelial cells in the present study, using viral transfection of shRNA. Unlike in Pkd1-/- cells, knockdown of PKD2 lowered basal Ca2+ and augmented store-operated Ca2+ entry, which was both independent of TMEM16A. However, disease causing purinergic Ca2+ store release was enhanced, similar to that observed in Pkd1-/- renal epithelial cells. The present data suggest pharmacological inhibition of TMEM16A as a treatment in ADPKD caused by mutations in both PKD1 and PKD2.

Keywords:  ADPKD; Anoctamin 1; Ca(2+) activated Cl(−) channel; Pkd1; Pkd2; Renal cysts; TMEM16A

DOI:  https://doi.org/10.1016/j.ceca.2021.102413
Biochem J. 2021 Apr 30. 478(8): 1617-1629

Kinase-anchoring proteins in ciliary signal transduction.

Janani Gopalan, Linda Wordeman, John D Scott.

  Historically, the diffusion of chemical signals through the cell was thought to occur within a cytoplasmic soup bounded by the plasma membrane. This theory was predicated on the notion that all regulatory enzymes are soluble and moved with a Brownian motion. Although enzyme compartmentalization was initially rebuffed by biochemists as a 'last refuge of a scoundrel', signal relay through macromolecular complexes is now accepted as a fundamental tenet of the burgeoning field of spatial biology. A-Kinase anchoring proteins (AKAPs) are prototypic enzyme-organizing elements that position clusters of regulatory proteins at defined subcellular locations. In parallel, the primary cilium has gained recognition as a subcellular mechanosensory organelle that amplifies second messenger signals pertaining to metazoan development. This article highlights advances in our understanding of AKAP signaling within the primary cilium and how defective ciliary function contributes to an increasing number of diseases known as ciliopathies.

Keywords:  AKAP; cilia; protein kinase A

DOI:  https://doi.org/10.1042/BCJ20200869
Am J Physiol Lung Cell Mol Physiol. 2021 Apr 28.

Rho/ROCK-MYOCD in regulating airway smooth muscle growth and remodeling.

Qin Yang, Wei Shi.

  Abnormal airway remodeling is a common pathologic change seen in chronic respiratory diseases. Altered proliferation and differentiation of airway smooth muscle cells (ASMCs) are the major component of airway remodeling, and the resultant structural abnormalities are difficult to restore. Understanding of airway smooth muscle regulation is urgently needed in order to identify potential intervention targets. MYOCD (or Myocardin) and Myocardin related transcription factors (MRTFs) are key co-transcription factors in muscle growth, which have not been extensively investigated in airway smooth muscle cells. In addition, the RhoA/ROCK signaling pathway is known to play an important role in airway remodeling partly through regulating the proliferation and differentiation of ASMCs, which may be connected with MYOCD/MRTF co-transcription factors. This minireview focuses on this newly recognized and potentially important RhoA/ROCK-MYOCD/MRTFs pathway in controlling airway smooth muscle growth and remodeling.

Keywords:  Airway; MYOCD; Smooth muscle cells

DOI:  https://doi.org/10.1152/ajplung.00034.2021
Development. 2021 May 01. pii: dev195552. [Epub ahead of print]148(9):

Hedgehog signaling and the primary cilium: implications for spatial and temporal constraints on signaling.

Emily K Ho, Tim Stearns.

  The mechanisms of vertebrate Hedgehog signaling are linked to the biology of the primary cilium, an antenna-like organelle that projects from the surface of most vertebrate cell types. Although the advantages of restricting signal transduction to cilia are often noted, the constraints imposed are less frequently considered, and yet they are central to how Hedgehog signaling operates in developing tissues. In this Review, we synthesize current understanding of Hedgehog signal transduction, ligand secretion and transport, and cilia dynamics to explore the temporal and spatial constraints imposed by the primary cilium on Hedgehog signaling in vivo.

Keywords:  Cell cycle; Cilia; Hedgehog; Signaling

DOI:  https://doi.org/10.1242/dev.195552
J Biomed Sci. 2021 Apr 28. 28(1): 33

RhoA: a dubious molecule in cardiac pathophysiology.

Lucia Sophie Kilian, Jakob Voran, Derk Frank, Ashraf Yusuf Rangrez.

  The Ras homolog gene family member A (RhoA) is the founding member of Rho GTPase superfamily originally studied in cancer cells where it was found to stimulate cell cycle progression and migration. RhoA acts as a master switch control of actin dynamics essential for maintaining cytoarchitecture of a cell. In the last two decades, however, RhoA has been coined and increasingly investigated as an essential molecule involved in signal transduction and regulation of gene transcription thereby affecting physiological functions such as cell division, survival, proliferation and migration. RhoA has been shown to play an important role in cardiac remodeling and cardiomyopathies; underlying mechanisms are however still poorly understood since the results derived from in vitro and in vivo experiments are still inconclusive. Interestingly its role in the development of cardiomyopathies or heart failure remains largely unclear due to anomalies in the current data available that indicate both cardioprotective and deleterious effects. In this review, we aimed to outline the molecular mechanisms of RhoA activation, to give an overview of its regulators, and the probable mechanisms of signal transduction leading to RhoA activation and induction of downstream effector pathways and corresponding cellular responses in cardiac (patho)physiology. Furthermore, we discuss the existing studies assessing the presented results and shedding light on the often-ambiguous data. Overall, we provide an update of the molecular, physiological and pathological functions of RhoA in the heart and its potential in cardiac therapeutics.

Keywords:  Actin dynamics; Cardiac pathophysiology; Cell proliferation; Rho GTPase; Signal transduction

DOI:  https://doi.org/10.1186/s12929-021-00730-w
iScience. 2021 Apr 23. 24(4): 102363

AMPKα2 activation by an energy-independent signal ensures chromosomal stability during mitosis.

Jianlin Lu, Yuanyuan Huang, Li Zhan, Ming Wang, Leilei Xu, McKay Mullen, Jianye Zang, Guowei Fang, Zhen Dou, Xing Liu, Wei Liu, Minerva Garcia-Barrio, Xuebiao Yao.

  AMP-activated protein kinase (AMPK) senses energy status and impacts energy-consuming events by initiating metabolism regulatory signals in cells. Accumulating evidences suggest a role of AMPK in mitosis regulation, but the mechanism of mitotic AMPK activation and function remains elusive. Here we report that AMPKα2, but not AMPKα1, is sequentially phosphorylated and activated by CDK1 and PLK1, which enables AMPKα2 to accurately guide chromosome segregation in mitosis. Phosphorylation at Thr485 by activated CDK1-Cyclin B1 brings the ST-stretch of AMPKα2 to the Polo box domain of PLK1 for subsequent Thr172 phosphorylation by PLK1. Inserting of the AMPKα2 ST-stretch into AMPKα1, which lacks the ST-stretch, can correct mitotic chromosome segregation defects in AMPKα2-depleted cells. These findings uncovered a specific signaling cascade integrating sequential phosphorylation by CDK1 and PLK1 of AMPKα2 with mitosis to maintain genomic stability, thus defining an isoform-specific AMPKα2 function, which will facilitate future research on energy sensing in mitosis.

Keywords:  Biological Sciences; Cell; Cell Biology

DOI:  https://doi.org/10.1016/j.isci.2021.102363
Biology (Basel). 2021 Apr 07. pii: 304. [Epub ahead of print]10(4):

New Insights into Cellular Functions of Nuclear Actin.

Malgorzata Kloc, Priyanka Chanana, Nicole Vaughn, Ahmed Uosef, Jacek Z Kubiak, Rafik M Ghobrial.

  Actin is one of the most abundant proteins in eukaryotic cells. There are different pools of nuclear actin often undetectable by conventional staining and commercial antibodies used to identify cytoplasmic actin. With the development of more sophisticated imaging and analytical techniques, it became clear that nuclear actin plays a crucial role in shaping the chromatin, genomic, and epigenetic landscape, transcriptional regulation, and DNA repair. This multifaceted role of nuclear actin is not only important for the function of the individual cell but also for the establishment of cell fate, and tissue and organ differentiation during development. Moreover, the changes in the nuclear, chromatin, and genomic architecture are preamble to various diseases. Here, we discuss some of the newly described functions of nuclear actin.

Keywords:  F actin; G actin; RhoA; chromatin remodeling; intranuclear rods; nuclear actin; nuclear architecture

DOI:  https://doi.org/10.3390/biology10040304
Int J Mol Sci. 2021 Apr 01. pii: 3656. [Epub ahead of print]22(7):

Role of Rho GTPase Interacting Proteins in Subcellular Compartments of Podocytes.

Kana Asano-Matsuda, Sajida Ibrahim, Tomoko Takano, Jun Matsuda.

  The first step of urine formation is the selective filtration of the plasma into the urinary space at the kidney structure called the glomerulus. The filtration barrier of the glomerulus allows blood cells and large proteins such as albumin to be retained while eliminating the waste products of the body. The filtration barrier consists of three layers: fenestrated endothelial cells, glomerular basement membrane, and podocytes. Podocytes are specialized epithelial cells featured by numerous, actin-based projections called foot processes. Proteins on the foot process membrane are connected to the well-organized intracellular actin network. The Rho family of small GTPases (Rho GTPases) act as intracellular molecular switches. They tightly regulate actin dynamics and subsequent diverse cellular functions such as adhesion, migration, and spreading. Previous studies using podocyte-specific transgenic or knockout animal models have established that Rho GTPases are crucial for the podocyte health and barrier function. However, little attention has been paid regarding subcellular locations where distinct Rho GTPases contribute to specific functions. In the current review, we discuss cellular events involving the prototypical Rho GTPases (RhoA, Rac1, and Cdc42) in podocytes, with particular focus on the subcellular compartments where the signaling events occur. We also provide our synthesized views of the current understanding and propose future research directions.

Keywords:  Cdc42; Rac1; Rho GTPase; RhoA; podocyte

DOI:  https://doi.org/10.3390/ijms22073656
Animals (Basel). 2021 Apr 09. pii: 1073. [Epub ahead of print]11(4):

Prostaglandin F2 Alpha Triggers the Disruption of Cell Adhesion with Cytokeratin and Vimentin in Bovine Luteal Theca Cells.

Sang-Hee Lee, Seunghyung Lee.

  Intermediate filaments (IFs) maintain cell-cell adhesions and are involved in diverse cellular processes such as cytokinesis, cell migration and the maintenance of cell structure. In this study, we investigated the influence of prostaglandin F2 alpha (PGF2α) on cytokeratin and vimentin IFs, Rho-associated protein kinase (ROCK), and cell-cell adhesion in bovine luteal theca cells (LTCs). The luteal cells were isolated from bovine corpus luteum (CL), and the LTCs were treated with 0, 0.01, 0.1 and 1.0 mM PGF2α. Cytokeratin, vimentin and desmoplakin proteins were disrupted and the ROCK protein was significantly increased in PGF2α-treated LTCs. In addition, cell-cell adhesion was significantly (p < 0.05) decreased in the PGF2α-induced LTCs compared to control group (0 mM PGF2α). In conclusion, PGF2α affected the adhesion of cell to cell via disruption of desmoplakin, cytokeratin and vimentin, additionally increasing ROCK in bovine LTCs. These results may provide a better understanding of the mechanism of bovine CL regression.

Keywords:  cell adhesion; corpus luteum; luteolysis; ovary; prostaglandin F2 alpha

DOI:  https://doi.org/10.3390/ani11041073
J Clin Med. 2021 Apr 07. pii: 1556. [Epub ahead of print]10(8):

Severity and Duration of Acute Kidney Injury and Chronic Kidney Disease after Cardiac Surgery.

Suk Hyung Choe, Hyeyeon Cho, Jinyoung Bae, Sang-Hwan Ji, Hyun-Kyu Yoon, Ho-Jin Lee, Ji-Hyun Lee, Jin-Tae Kim, Won Ho Kim.

  We aimed to evaluate whether the duration and stage of acute kidney injury (AKI) are associated with the occurrence of chronic kidney disease (CKD) in patients undergoing cardiac or thoracic aortic surgery. A total of 2009 cases were reviewed. The patients with postoperative AKI stage 1 and higher stage were divided into transient (serum creatinine elevation ≤48 h) or persistent (>48 h) AKI, respectively. Estimated glomerular filtration rate (eGFR) values during three years after surgery were collected. Occurrence of new-onset CKD stage 3 or higher or all-cause mortality was determined as the primary outcome. Multivariable Cox regression and Kaplan-Meier survival analysis were performed. The Median follow-up of renal function after surgery was 32 months. The cumulative incidences of our primary outcome at one, two, and three years after surgery were 19.8, 23.7, and 26.1%. There was a graded significant association of AKI with new-onset CKD during three years after surgery, except for transient stage 1 AKI (persistent stage 1: HR 3.11, 95% CI 2.62-4.91; transient higher stage: HR 4.07, 95% CI 2.98-6.11; persistent higher stage: HR 13.36, 95% CI 8.22-18.72). There was a significant difference in survival between transient and persistent AKI at the same stage. During three years after cardiac surgery, there was a significant and graded association between AKI stages and the development of new-onset CKD, except for transient stage 1 AKI. This association was stronger when AKI lasted more than 48 h at the same stage. Both duration and severity of AKI provide prognostic value to predict the development of CKD.

Keywords:  acute kidney injury; cardiac surgery; chronic kidney disease; creatinine; duration; risk factor

DOI:  https://doi.org/10.3390/jcm10081556
Nephrol Dial Transplant. 2020 Jul 10. pii: gfaa091. [Epub ahead of print]

Hypoxia in chronic kidney disease: towards a paradigm shift?

Anna Faivre, Carsten C Scholz, Sophie de Seigneux.

  Chronic kidney disease (CKD) is defined as an alteration of kidney structure and/or function lasting for >3 months [1]. CKD affects 10% of the general adult population and is responsible for large healthcare costs [2]. Since the end of the last century, the role of hypoxia in CKD progression has controversially been discussed. To date, there is evidence of the presence of hypoxia in late-stage renal disease, but we lack time-course evidence, stage correlation and also spatial co-localization with fibrotic lesions to ensure its causative role. The classical view of hypoxia in CKD progression is that it is caused by peritubular capillary alterations, renal anaemia and increased oxygen consumption regardless of the primary injury. In this classical view, hypoxia is assumed to further induce pro-fibrotic and pro-inflammatory responses, as well as oxidative stress, leading to CKD worsening as part of a vicious circle. However, recent investigations tend to question this paradigm, and both the presence of hypoxia and its role in CKD progression are still not clearly demonstrated. Hypoxia-inducible factor (HIF) is the main transcriptional regulator of the hypoxia response. Genetic HIF modulation leads to variable effects on CKD progression in different murine models. In contrast, pharmacological modulation of the HIF pathway [i.e. by HIF hydroxylase inhibitors (HIs)] appears to be generally protective against fibrosis progression experimentally. We here review the existing literature on the role of hypoxia, the HIF pathway and HIF HIs in CKD progression and summarize the evidence that supports or rejects the hypoxia hypothesis, respectively.

Keywords:  HIF; PHI; chronic kidney disease; hydroxylase inhibitor; hypoxia

DOI:  https://doi.org/10.1093/ndt/gfaa091
Cell Death Dis. 2021 Apr 30. 12(5): 432

Permissive effect of GSK3β on profibrogenic plasticity of renal tubular cells in progressive chronic kidney disease.

Bohan Chen, Pei Wang, Xianhui Liang, Chunming Jiang, Yan Ge, Lance D Dworkin, Rujun Gong.

Renal tubular epithelial cells (TECs) play a key role in renal fibrogenesis. After persistent injuries that are beyond self-healing capacity, TECs will dedifferentiate, undergo growth arrest, convert to profibrogenic phenotypes, and resort to maladaptive plasticity that ultimately results in renal fibrosis. Evidence suggests that glycogen synthase kinase (GSK) 3β is centrally implicated in kidney injury. However, its role in renal fibrogenesis is obscure. Analysis of publicly available kidney transcriptome database demonstrated that patients with progressive chronic kidney disease (CKD) exhibited GSK3β overexpression in renal tubulointerstitium, in which the predefined hallmark gene sets implicated in fibrogenesis were remarkably enriched. In vitro, TGF-β1 treatment augmented GSK3β expression in TECs, concomitant with dedifferentiation, cell cycle arrest at G2/M phase, excessive accumulation of extracellular matrix, and overproduction of profibrotic cytokines like PAI-1 and CTGF. All these profibrogenic phenotypes were largely abrogated by GSK3β inhibitors or by ectopic expression of a dominant-negative mutant of GSK3β but reinforced in cells expressing the constitutively active mutant of GSK3β. Mechanistically, GSK3β suppressed, whereas inhibiting GSK3β facilitated, the activity of cAMP response element-binding protein (CREB), which competes for CREB-binding protein, a transcriptional coactivator essential for TGF-β1/Smad signaling pathway to drive TECs profibrogenic plasticity. In vivo, in mice with folic acid-induced progressive CKD, targeting of GSK3β in renal tubules via genetic ablation or by microdose lithium mitigated the profibrogenic plasticity of TEC, concomitant with attenuated interstitial fibrosis and tubular atrophy. Collectively, GSK3β is likely a pragmatic therapeutic target for averting profibrogenic plasticity of TECs and improving renal fibrosis.

DOI: https://doi.org/10.1038/s41419-021-03709-5
Int J Mol Sci. 2021 Apr 28. pii: 4645. [Epub ahead of print]22(9):

Myocardial Hypertrophy and Fibrosis Are Associated with Cardiomyocyte Beta-Catenin and TRPC6/Calcineurin/NFAT Signaling in Spontaneously Hypertensive Rats with 5/6 Nephrectomy.

Evdokia Bogdanova, Olga Beresneva, Olga Galkina, Irina Zubina, Galina Ivanova, Marina Parastaeva, Natalia Semenova, Vladimir Dobronravov.

   BACKGROUND: Arterial hypertension (AH) is associated with heart and chronic kidney disease (CKD). However, the precise mechanisms of myocardial remodeling (MR) in the settings of CKD remain elusive. We hypothesized that TRPC6, calcineurin/NFAT, and Wnt/β-catenin signaling pathways are involved in the development of MR in the background of CKD and AH.
METHODS: Early CKD was induced by performing a 5/6 nephrectomy (5/6NE) in spontaneously hypertensive rats (SHR-NE). Sham-operated (SO) SHR (SHR-SO) and Wistar Kyoto (WKY-SO) rats served as controls. Systolic blood pressure (SBP), heart rate, myocardial mass index (MMI), serum creatinine, cardiomyocyte diameter (dCM), myocardial fibrosis (MF), serum and kidney α-Klotho levels, myocardial expression of calcineurin (CaN), TRPC6, and β-catenin were measured two months after 5/6NE or SO.
RESULTS: NE-induced kidney dysfunction corresponded to mild-to-moderate human CKD and was associated with an increase in FGF23 and a decrease in renal α-Klotho. The levels of SBP, MMI, dCM, and MF were higher in SHRs compared to WKY-SO as well as in SHR-NE vs. SHR-SO. The MR was associated with increased cardiomyocyte expression of CaN/NFAT and β-catenin along with its intracellular re-distribution. TRPC6 protein levels were substantially elevated in both SHR groups with higher Trpc6 mRNA expression in SHR-NE.
CONCLUSIONS: The Wnt/β-catenin and TRPC6/CaN/NFAT hypertrophic signaling pathways seem to be involved in myocardial remodeling in the settings of AH and CKD and might be mediated by FGF23 and α-Klotho axis.

Keywords:  Klotho; NFAT; TRPC6; arterial hypertension; calcineurin A; calcineurin B; cardiac remodeling; chronic kidney disease; fibroblast growth factor 23; parathyroid hormone; β-catenin

DOI:  https://doi.org/10.3390/ijms22094645
Theranostics. 2021 ;11(12): 6090-6104

Mechanically induced integrin ligation mediates intracellular calcium signaling with single pulsating cavitation bubbles.

Fenfang Li, Tae Hyun Park, George Sankin, Christopher Gilchrist, Defei Liao, Chon U Chan, Zheng Mao, Brenton D Hoffman, Pei Zhong.

  Therapeutic ultrasound or shockwave has shown its great potential to stimulate neural and muscle tissue, where cavitation microbubble induced Ca2+ signaling is believed to play an important role. However, the pertinent mechanisms are unknown, especially at the single-cell level. Particularly, it is still a major challenge to get a comprehensive understanding of the effect of potential mechanosensitive molecular players on the cellular responses, including mechanosensitive ion channels, purinergic signaling and integrin ligation by extracellular matrix. Methods: Here, laser-induced cavitation microbubble was used to stimulate individual HEK293T cells either genetically knocked out or expressing Piezo1 ion channels with different normalized bubble-cell distance. Ca2+ signaling and potential membrane poration were evaluated with a real-time fluorescence imaging system. Integrin-binding microbeads were attached to the apical surface of the cells at mild cavitation conditions, where the effect of Piezo1, P2X receptors and integrin ligation on single cell intracellular Ca2+ signaling was assessed. Results: Ca2+ responses were rare at normalized cell-bubble distances that avoided membrane poration, even with overexpression of Piezo1, but could be increased in frequency to 42% of cells by attaching integrin-binding beads. We identified key molecular players in the bead-enhanced Ca2+ response: increased integrin ligation by substrate ECM triggered ATP release and activation of P2X-but not Piezo1-ion channels. The resultant Ca2+ influx caused dynamic changes in cell spread area. Conclusion: This approach to safely eliciting a Ca2+ response with cavitation microbubbles and the uncovered mechanism by which increased integrin-ligation mediates ATP release and Ca2+ signaling will inform new strategies to stimulate tissues with ultrasound and shockwaves.

Keywords:  ATP-gated P2X channel.; Intracellular calcium signaling; Piezo1; cavitation bioeffects; integrin ligation

DOI:  https://doi.org/10.7150/thno.56813
Int J Mol Sci. 2021 Apr 27. pii: 4550. [Epub ahead of print]22(9):

Simulated Microgravity Inhibits the Proliferation of Chang Liver Cells by Attenuation of the Major Cell Cycle Regulators and Cytoskeletal Proteins.

Chi Nguyen Quynh Ho, Minh Thi Tran, Chung Chinh Doan, Son Nghia Hoang, Diem Hong Tran, Long Thanh Le.

  Simulated microgravity (SMG) induced the changes in cell proliferation and cytoskeleton organization, which plays an important factor in various cellular processes. The inhibition in cell cycle progression has been considered to be one of the main causes of proliferation inhibition in cells under SMG, but their mechanisms are still not fully understood. This study aimed to evaluate the effects of SMG on the proliferative ability and cytoskeleton changes of Chang Liver Cells (CCL-13). CCL-13 cells were induced SMG by 3D clinostat for 72 h, while the control group were treated in normal gravity at the same time. The results showed that SMG reduced CCL-13 cell proliferation by an increase in the number of CCL-13 cells in G0/G1 phase. This cell cycle phase arrest of CCL-13 cells was due to a downregulation of cell cycle-related proteins, such as cyclin A1 and A2, cyclin D1, and cyclin-dependent kinase 6 (Cdk6). SMG-exposed CCL-13 cells also exhibited a downregulation of α-tubulin 3 and β-actin which induced the cytoskeleton reorganization. These results suggested that the inhibited proliferation of SMG-exposed CCL-13 cells could be associate with the attenuation of major cell cycle regulators and main cytoskeletal proteins.

Keywords:  Chang Liver Cells; cell cycle regulators; cytoskeleton; proliferation; simulated microgravity

DOI:  https://doi.org/10.3390/ijms22094550
Cell. 2021 Apr 27. pii: S0092-8674(21)00435-9. [Epub ahead of print]

Splice site m6A methylation prevents binding of U2AF35 to inhibit RNA splicing.

Mateusz Mendel, Kamila Delaney, Radha Raman Pandey, Kuan-Ming Chen, Joanna M Wenda, Cathrine Broberg Vågbø, Florian A Steiner, David Homolka, Ramesh S Pillai.

  The N6-methyladenosine (m6A) RNA modification is used widely to alter the fate of mRNAs. Here we demonstrate that the C. elegans writer METT-10 (the ortholog of mouse METTL16) deposits an m6A mark on the 3' splice site (AG) of the S-adenosylmethionine (SAM) synthetase pre-mRNA, which inhibits its proper splicing and protein production. The mechanism is triggered by a rich diet and acts as an m6A-mediated switch to stop SAM production and regulate its homeostasis. Although the mammalian SAM synthetase pre-mRNA is not regulated via this mechanism, we show that splicing inhibition by 3' splice site m6A is conserved in mammals. The modification functions by physically preventing the essential splicing factor U2AF35 from recognizing the 3' splice site. We propose that use of splice-site m6A is an ancient mechanism for splicing regulation.

Keywords:  3' splice site; METT-10; METTL16; SAM homeostasis; SAM synthetase; U2AF35/65; U6 snRNA; m(6)A; spermatogenesis; splicing

DOI:  https://doi.org/10.1016/j.cell.2021.03.062
Int J Mol Sci. 2021 Apr 25. pii: 4473. [Epub ahead of print]22(9):

The Roles of Calmodulin and CaMKII in Cx36 Plasticity.

Georg R Zoidl, David C Spray.

  Anatomical and electrophysiological evidence that gap junctions and electrical coupling occur between neurons was initially confined to invertebrates and nonmammals and was thought to be a primitive form of synaptic transmission. More recent studies revealed that electrical communication is common in the mammalian central nervous system (CNS), often coexisting with chemical synaptic transmission. The subsequent progress indicated that electrical synapses formed by the gap junction protein connexin-36 (Cx36) and its paralogs in nonmammals constitute vital elements in mammalian and fish synaptic circuitry. They govern the collective activity of ensembles of coupled neurons, and Cx36 gap junctions endow them with enormous adaptive plasticity, like that seen at chemical synapses. Moreover, they orchestrate the synchronized neuronal network activity and rhythmic oscillations that underlie the fundamental integrative processes, such as memory and learning. Here, we review the available mechanistic evidence and models that argue for the essential roles of calcium, calmodulin, and the Ca2+/calmodulin-dependent protein kinase II in integrating calcium signals to modulate the strength of electrical synapses through interactions with the gap junction protein Cx36.

Keywords:  CaMKII; calcium; calmodulin; connexin-36 (Cx36); electrical synapse; plasticity; tubulin

DOI:  https://doi.org/10.3390/ijms22094473