bims-bicyki 2021-11-21 papers

bims-bicyki

Biomed News

on Bicaudal-C1 and interactors in cystic kidney disease

Issue of 2021–11–21
eleven papers selected by
Céline Gagnieux, École Polytechnique Fédérale de Lausanne

Right Ventricular Blood Cyst in an Adult Autosomal Dominant Polycystic Kidney Disease Patient:<br /> An Unusual Association.
The Impact of COVID-19 on Patients With ADPKD.
Nitric oxide could promote development of Barrett's esophagus by S-nitrosylation-induced inhibition of Rho-ROCK signaling in esophageal fibroblasts.
CEP97 phosphorylation by Dyrk1a is critical for centriole separation during multiciliogenesis.
Ciliary Type III Adenylyl Cyclase in the VMH Is Crucial for High-Fat Diet-Induced Obesity Mediated by Autophagy.
Kinesin family member 2A acts as a potential prognostic marker and treatment target via interaction with PI3K/AKT and RhoA/ROCK pathways in acute myeloid leukemia.
Conditional deletion of ROCK2 induces anxiety-like behaviors and alters dendritic spine density and morphology on CA1 pyramidal neurons.
Dimeric Rhodopsin R135L Mutant-Transducin-like Complex Sheds Light on Retinitis Pigmentosa Misfunctions.
Gα13 Mediates Transendothelial Migration of Neutrophils by Promoting Integrin-Dependent Motility without Affecting Directionality.
Modeling pulmonary cystic fibrosis in a human lung airway-on-a-chip: Cystic fibrosis airway chip.
Optimized expression of recombinant human NIMA-related kinase 7 (NEK7) with a higher purity in Escherichia coli.

J Coll Physicians Surg Pak. 2021 Dec;31(12): 1500-1502

Right Ventricular Blood Cyst in an Adult Autosomal Dominant Polycystic Kidney Disease Patient:<br /> An Unusual Association.

Ersin Cagri Simsek, Dilek Oncel, Erdem Ozel.

Blood cysts of the heart are unusual primary cardiac tumor-like masses, which are usually located on the cardiac valves. These are very rare in adults, especially in a non-valvular location. Autosomal-dominant polycystic kidney disease (ADPKD) typically presents with multiple bilateral renal cysts, resulting in chronic kidney disease. Whilst many of the extra-renal manifestations of ADPKD are well-documented, associated cardiac masses are extremely rare: and cardiac blood cyst has not been reported in a patient with APKD to date. We present a 57-year man with a history of ADPKD and end-stage renal disease with a 2-cm-diameter right ventricular blood cyst, which was detected on multimodality imaging. Key words: Autosomal dominant polycystic kidney disease, Cardiac blood cyst, Extra-renal manifestations.

DOI: https://doi.org/10.29271/jcpsp.2021.12.1500
Can J Kidney Health Dis. 2021 ;8 20543581211056479

The Impact of COVID-19 on Patients With ADPKD.

Meherzad Kutky, Erin Cross, Darin J Treleaven, Ahsan Alam, Matthew B Lanktree.

   Purpose of review: Patients with autosomal dominant polycystic kidney disease (ADPKD) have kidney cysts and kidney enlargement decades before progressing to advanced chronic kidney disease (CKD), meaning patients live most of their adult life with a chronic medical condition. The coronavirus disease 2019 (COVID-19) pandemic has created common questions among patients with ADPKD. In this review, we discuss COVID-19 concerns centered around a patient with a common clinical vignette.
Sources of information: We performed PubMed and Google scholar searches for English, peer-reviewed studies related to "COVID-19," "ADPKD," "CKD," "tolvaptan," "angiotensin-converting enzyme inhibitors" (ACEi), "angiotensin receptor blockers" (ARB), and "vaccination." We also evaluated transplant data provided by the Ontario Trillium Gift of Life Network.
Methods: Following an assessment of available literature, this narrative review addresses common questions of patients with ADPKD in the context of the COVID-19 pandemic.
Key findings: Data regarding the risk of developing COVID-19 and the risk of adverse COVID-19 outcomes in patients with ADPKD remain limited, but patients with ADPKD with impaired estimated glomerular filtration rate (eGFR), kidney transplants, or on dialysis are likely at similar increased risk as those with generally defined CKD. We provide strategies to improve virtual care, which is likely to persist after the pandemic. Current evidence suggests ACEi, ARB, and tolvaptan treatment should be continued unless contraindicated due to severe illness. When available, and in the absence of a severe allergy, vaccination is recommended for all patients with ADPKD.
Limitations: This narrative review is limited by a paucity of high-quality data on COVID-19 outcomes in patients specifically with ADPKD.
Implications: Patients with ADPKD who have developed advanced CKD, require dialysis, or who have received a kidney transplant are at elevated risk of COVID-19 complications.

Keywords:  COVID-19; autosomal dominant polycystic kidney disease (ADPKD); transplantation; vaccine

DOI:  https://doi.org/10.1177/20543581211056479
Am J Physiol Gastrointest Liver Physiol. 2021 Nov 17.

Nitric oxide could promote development of Barrett's esophagus by S-nitrosylation-induced inhibition of Rho-ROCK signaling in esophageal fibroblasts.

Taku Fujiya, Kiyotaka Asanuma, Tomoyuki Koike, Tomoki Okata, Masahiro Saito, Naoki Asano, Akira Imatani, Atsushi Masamune.

  Barrett's esophagus arises in the process of wound healing in distal esophageal epithelium damaged by gastroesophageal reflux disease. Differentiation of fibroblast into myofibroblasts, a smooth muscle cell-like phenotype and tissue contraction are crucial processes in wound healing. No study has evaluated mechanism by which luminal esophageal nitric oxide (NO) affect Rho-associated coiled-coil forming protein kinase (Rho-ROCK) signaling pathway, a key factor of tissue contraction, in stromal fibroblasts to develop Barrett's esophagus. Using esophageal fibroblasts, we performed collagen-based cell contraction assays and evaluated influence of Rho-ROCK signaling in the exposure to acidic bile salts and NOC-9, which is an NO donor. We found that enhanced cell contraction induced by acidic bile salts was inhibited by NO, accompanied by decrease in phosphorylated myosin light chain expression and stress fiber formation. NO directly S-nitrosylated GTP-RhoA and consequently blocked Rho-ROCK signaling. Moreover, exposure to NO and Y27632, a Rho-ROCK signaling inhibitor, decreased a-SMA expression and increased bone morphogenetic protein 4 (BMP4) expression and secretion. These findings could account for the increased expression of BMP4 in the columnar epithelial cells and stromal fibroblasts in human Barrett's esophagus. NO could impair wound healing by blocking the Rho-ROCK signaling pathway and promote development of Barrett's esophagus.

Keywords:  Barrett's esophagus; Rho-ROCK signaling; S-nitrosylation; nitric oxide; tissue contration

DOI:  https://doi.org/10.1152/ajpgi.00124.2021
J Cell Biol. 2022 Jan 03. pii: e202102110. [Epub ahead of print]221(1):

CEP97 phosphorylation by Dyrk1a is critical for centriole separation during multiciliogenesis.

Moonsup Lee, Kunio Nagashima, Jaeho Yoon, Jian Sun, Ziqiu Wang, Christina Carpenter, Hyun-Kyung Lee, Yoo-Seok Hwang, Christopher J Westlake, Ira O Daar.

Proper cilia formation in multiciliated cells (MCCs) is necessary for appropriate embryonic development and homeostasis. Multicilia share many structural characteristics with monocilia and primary cilia, but there are still significant gaps in our understanding of the regulation of multiciliogenesis. Using the Xenopus embryo, we show that CEP97, which is known as a negative regulator of primary cilia formation, interacts with dual specificity tyrosine phosphorylation regulated kinase 1A (Dyrk1a) to modulate multiciliogenesis. We show that Dyrk1a phosphorylates CEP97, which in turn promotes the recruitment of Polo-like kinase 1 (Plk1), which is a critical regulator of MCC maturation that functions to enhance centriole disengagement in cooperation with the enzyme Separase. Knockdown of either CEP97 or Dyrk1a disrupts cilia formation and centriole disengagement in MCCs, but this defect is rescued by overexpression of Separase. Thus, our study reveals that Dyrk1a and CEP97 coordinate with Plk1 to promote Separase function to properly form multicilia in vertebrate MCCs.

DOI: https://doi.org/10.1083/jcb.202102110
Adv Sci (Weinh). 2021 Nov 16. e2102568

Ciliary Type III Adenylyl Cyclase in the VMH Is Crucial for High-Fat Diet-Induced Obesity Mediated by Autophagy.

Dong Yang, Xiangbo Wu, Weina Wang, Yanfen Zhou, Zhenshan Wang.

  Neuronal primary cilia are crucial for body weight maintenance. Type III adenylyl cyclase (AC3) is abundantly enriched in neuronal cilia, and mice with global AC3 ablation are obese. However, whether AC3 regulates body weight through its ciliary expression and the mechanism underlying this potential regulation are not clear. In this study, humanized AC3 knock-in mice that are resistant to high-fat diet (HFD)-induced obesity are generated, and increases in the number and length of cilia in the ventromedial hypothalamus (VMH) are shown. It is demonstrated that mice with specifically knocked down ciliary AC3 expression in the VMH show pronounced HFD-induced obesity. In addition, in vitro and in vivo analyses of the VMH show that ciliary AC3 regulates autophagy by binding an autophagy-related gene, gamma-aminobutyric acid A receptor-associated protein (GABARAP). Mice with GABARAP knockdown in the VMH exhibit exacerbated HFD-induced obesity. Overall, the findings may reveal a potential mechanism by which ciliary AC3 expression regulates body weight in the mouse VMH.

Keywords:  autophagy; cilia; obesity; type III adenylyl cyclase; ventromedial hypothalamus

DOI:  https://doi.org/10.1002/advs.202102568
Oncol Rep. 2022 Jan;pii: 18. [Epub ahead of print]47(1):

Kinesin family member 2A acts as a potential prognostic marker and treatment target via interaction with PI3K/AKT and RhoA/ROCK pathways in acute myeloid leukemia.

Xinglin Liang, Ruixiang Xia.

  KIF2A has been shown to be involved in the regulation of AML pathology, however, the mechanistic role of KIF2A in AML has not been fully identified. The present study aimed to identify the underlying mechanism of KIF2A regulation of AML cell function and chemosensitivity. A total of 58 patients with AML and 30 healthy subjects were enrolled for clinical analysis. AML cells (KG‑1 and Kasumi‑1) were transfected with KIF2A or control small interfering (si)RNA. PI3K/AKT pathway activator (740 Y‑P) and RhoA overexpression plasmid were added to rescue the effect of KIF2A siRNA. Cell proliferation, apoptosis, chemosensitivity to ADR and AraC, expression levels of mRNA/proteins associated with PI3K/AKT and RhoA/ROCK pathways were measured by Cell Counting Kit‑8, flow cytometry, reverse transcription‑quantitative PCR and western blotting. KIF2A was overexpressed, and correlated with higher levels of bone marrow blast, poor risk classification, lower treatment response and unfavorable survival profile in patients with AML. KIF2A siRNA inhibited proliferation but enhanced apoptosis and chemosensitivity to ADR and AraC in KG‑1 and Kasumi‑1 cells, which also inactivated PI3K/AKT and RhoA/ROCK pathways. Subsequent rescue experiments showed that 740 Y‑P and RhoA overexpression plasmid promoted cell survival and decreased chemosensitivity, which reversed the effect of KIF2A siRNA in KG‑1 and Kasumi‑1 cells. KIF2A was correlated with worse clinical features and survival in patients with AML; its knockdown promoted apoptosis and chemosensitivity by inactivating PI3K/AKT and RhoA/ROCK signaling pathways in AML cells. These data suggested KIF2A may be a potential prognostic marker and treatment target for AML management.

Keywords:  PI3K/AKT pathway; acute myeloid leukemia; cell function; chemosensitivity; kinesin family member 2A; ras homolog family member A/rho‑associated coiled‑coil containing protein kinase pathway

DOI:  https://doi.org/10.3892/or.2021.8229
Mol Brain. 2021 Nov 18. 14(1): 169

Conditional deletion of ROCK2 induces anxiety-like behaviors and alters dendritic spine density and morphology on CA1 pyramidal neurons.

Audrey J Weber, Ashley B Adamson, Kelsey M Greathouse, Julia P Andrade, Cameron D Freeman, Jung Vin Seo, Rosaria J Rae, Courtney K Walker, Jeremy H Herskowitz.

  Rho-associated kinase isoform 2 (ROCK2) is an attractive drug target for several neurologic disorders. A critical barrier to ROCK2-based research and therapeutics is the lack of a mouse model that enables investigation of ROCK2 with spatial and temporal control of gene expression. To overcome this, we generated ROCK2fl/fl mice. Mice expressing Cre recombinase in forebrain excitatory neurons (CaMKII-Cre) were crossed with ROCK2fl/fl mice (Cre/ROCK2fl/fl), and the contribution of ROCK2 in behavior as well as dendritic spine morphology in the hippocampus, medial prefrontal cortex (mPFC), and basolateral amygdala (BLA) was examined. Cre/ROCK2fl/fl mice spent reduced time in the open arms of the elevated plus maze and increased time in the dark of the light-dark box test compared to littermate controls. These results indicated that Cre/ROCK2fl/fl mice exhibited anxiety-like behaviors. To examine dendritic spine morphology, individual pyramidal neurons in CA1 hippocampus, mPFC, and the BLA were targeted for iontophoretic microinjection of fluorescent dye, followed by high-resolution confocal microscopy and neuronal 3D reconstructions for morphometry analysis. In dorsal CA1, Cre/ROCK2fl/fl mice displayed significantly increased thin spine density on basal dendrites and reduced mean spine head volume across all spine types on apical dendrites. In ventral CA1, Cre/ROCK2fl/fl mice exhibited significantly increased spine length on apical dendrites. Spine density and morphology were comparable in the mPFC and BLA between both genotypes. These findings suggest that neuronal ROCK2 mediates spine density and morphology in a compartmentalized manner among CA1 pyramidal cells, and that in the absence of ROCK2 these mechanisms may contribute to anxiety-like behaviors.

Keywords:  Amygdala; Dendritic spine; Hippocampus; Prefrontal cortex; ROCK2; Rho kinase

DOI:  https://doi.org/10.1186/s13041-021-00878-4
J Phys Chem B. 2021 Nov 18.

Dimeric Rhodopsin R135L Mutant-Transducin-like Complex Sheds Light on Retinitis Pigmentosa Misfunctions.

Diego M A Guérin, Ayelen Digilio, María Marta Branda.

Rhodopsin (RHO) is a light-sensitive pigment in the retina and the main prototypical protein of the G-protein-coupled receptor (GCPR) family. After receiving a light stimulus, RHO and its cofactor retinylidene undergo a series of structural changes that initiate an intricate transduction mechanism. Along with RHO, other partner proteins play key roles in the signaling pathway. These include transducin, a GTPase, kinases that phosphorylate RHO, and arrestin (Arr), which ultimately stops the signaling process and promotes RHO regeneration. A large number of RHO genetic mutations may lead to very severe retinal dysfunction and eventually to impaired dark adaptation disease called autosomal dominant retinitis pigmentosa (adRP). In this study, we used molecular dynamics (MD) simulations to evaluate the different behaviors of the dimeric form of wild-type RHO (WT dRHO) and its mutant at position 135 of arginine to leucine (dR135L), both in the free (noncomplexed) and in complex with the transducin-like protein (Gtl). Gtl is a heterotrimeric model composed of a mixture of human and bovine G proteins. Our calculations allow us to explain how the mutation causes structural changes in the RHO dimer and how this can affect the signal that transducin generates when it is bound to RHO. Moreover, the structural modifications induced by the R135L mutation can also account for other misfunctions observed in the up- and downstream signaling pathways. The mechanism of these dysfunctions, together with the transducin activity reduction, provides structure-based explanations of the impairment of some key processes that lead to adRP.

DOI: https://doi.org/10.1021/acs.jpcb.1c06348
J Immunol. 2021 Nov 19. pii: ji2001385. [Epub ahead of print]

Gα13 Mediates Transendothelial Migration of Neutrophils by Promoting Integrin-Dependent Motility without Affecting Directionality.

Claire W Chang, Ni Cheng, Yanyan Bai, Randal A Skidgel, Xiaoping Du.

Neutrophil migration requires β2 integrins and chemoattractant receptor signaling for motility and directionality. G protein subunit Gα13 can facilitate cell migration by mediating RhoA activation induced by G protein-coupled receptors. However, the possible role of Gα13-integrin interaction in migration is unclear. In this study, we show that Gα13 -/- neutrophils are deficient in transendothelial migration and migration on β2 integrin ligand ICAM-1. However, unlike G protein-coupled receptors and integrin inside-out signaling pathways, Gα13 is important in migration velocity and neutrophil spreading but not in directionality nor cell adhesion. Importantly, neutrophil recruitment in vivo was also inhibited in Gα13 -/- mice, suggesting the importance of Gα13 in transendothelial migration of neutrophils in vitro and in vivo. Furthermore, a synthetic peptide (MB2mP6) derived from the Gα13 binding site of β2 inhibited Gα13-β2 interaction and Gα13-mediated transient RhoA inhibition in neutrophils, suggesting that this peptide inhibited integrin outside-in signaling. MB2mP6 inhibited migration of control neutrophils through endothelial cell monolayers or ICAM-1-coated filters, but was without further effect on Gα13 -/- neutrophils. It also inhibited integrin-dependent neutrophil migration velocity without affecting directionality. In vivo, MB2mP6 markedly inhibited neutrophil infiltration into the cardiac tissues induced by ischemia/reperfusion injury. Thus, Gα13-dependent outside-in signaling enables integrin-dependent neutrophil motility without affecting directionality and may be a new therapeutic target for inhibiting neutrophil trafficking but not adhesion.

DOI: https://doi.org/10.4049/jimmunol.2001385
J Cyst Fibros. 2021 Nov 16. pii: S1569-1993(21)02106-8. [Epub ahead of print]

Modeling pulmonary cystic fibrosis in a human lung airway-on-a-chip: Cystic fibrosis airway chip.

Roberto Plebani, Ratnakar Potla, Mercy Soong, Haiqing Bai, Zohreh Izadifar, Amanda Jiang, Renee N Travis, Chaitra Belgur, Alexandre Dinis, Mark J Cartwright, Rachelle Prantil-Baun, Pawan Jolly, Sarah E Gilpin, Mario Romano, Donald E Ingber.

   BACKGROUND: Cystic fibrosis (CF) is a genetic disease caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR), which results in impaired airway mucociliary clearance, inflammation, infection, and respiratory insufficiency. The development of new therapeutics for CF are limited by the lack of reliable preclinical models that recapitulate the structural, immunological, and bioelectrical features of human CF lungs.
METHODS: We leveraged organ-on-a-chip technology to develop a microfluidic device lined by primary human CF bronchial epithelial cells grown under an air-liquid interface and interfaced with pulmonary microvascular endothelial cells (CF Airway Chip) exposed to fluid flow. The responses of CF and healthy Airway Chips were analyzed in the presence or absence of polymorphonuclear leukocytes (PMNs) and the bacterial pathogen, Pseudomonas aeruginosa.
RESULTS: The CF Airway Chip faithfully recapitulated many features of the human CF airways, including enhanced mucus accumulation, increased cilia density, and a higher ciliary beating frequency compared to chips lined by healthy bronchial epithelial cells. The CF chips also secreted higher levels of IL-8, which was accompanied by enhanced PMN adhesion to the endothelium and transmigration into the airway compartment. In addition, CF Airway Chips provided a more favorable environment for Pseudomonas aeruginosa growth, which resulted in enhanced secretion of inflammatory cytokines and recruitment of PMNs to the airway.
CONCLUSIONS: The human CF Airway Chip may provide a valuable preclinical tool for pathophysiology studies as well as for drug testing and personalized medicine.

Keywords:  Cystic fibrosis; Microfluidics; Neutrophils; Organ chip; Pseudomonas

DOI:  https://doi.org/10.1016/j.jcf.2021.10.004
Protein Pept Lett. 2021 Nov 17.

Optimized expression of recombinant human NIMA-related kinase 7 (NEK7) with a higher purity in Escherichia coli.

Xing-Jie Zhang, Ting-Ting Wang, Yu-Kun Pu, Lin Zeng, Rui-Han Zhang, Xiao-Li Li, Xu Ji, Wei-Lie Xiao.

   BACKGROUND: NIMA (never in mitosis, gene A) serine/threonine kinase 7 (NEK7) is a regulator of mitosis spindle in mammals and is considered as a drug target of inflammasome related inflammatory diseases. However, most commercially available or reported recombinant NEK7 proteins are either inactive or have low purity. These shortcomings limit the pharmacological studies and development of NEK7 inhibitors.
OBJECTIVE: To elucidate what causes the NEK7 low purity in E. coli, and optimize a protocol to improve the protein purity.
METHODS: A comparative study of expression full length NEK7 with an N-terminal His-tag or a Cterminal His-tag was performed. His-affinity resin, ion exchange and gel filtration chromatography were used to purify NEK7. The protein was identified by mass spectrometry. The activity and folding of NEK7 were evaluated by chemiluminescent assay and thermal shift assay.
RESULTS: Our results demonstrated that N-terminal tagged protein was toxic to E. coli, resulting in incomplete translated products. The C-terminal tagged NEK7-His6 had a much higher purity than that of an N-terminal tag. The Ni2+ resin one-step purification led to a purity of 91.7%, meeting the criteria of most kinase assays. With two-step and three-step procedures, the protein purities were 94.7% and ~100%, respectively. The NEK7 purified in this work maintained its kinase activity and correct conformation, and the compound-protein interaction ability.
CONCLUSION: Our optimized protocol could produce good purity of His tagged NEK7 in E. coli, and the kinase activity and biophysical characteristics of which are preserved.

Keywords:  NEK7; expression; high purity; histidine tag; kinase; purification

DOI:  https://doi.org/10.2174/0929866528666211118092410