bims-bicyki 2021-03-07 papers

bims-bicyki

Biomed News

on Bicaudal-C1 and interactors in cystic kidney disease

Issue of 2021‒03‒07
sixteen papers selected by
Céline Gagnieux
École Polytechnique Fédérale de Lausanne (EPFL)

Clinical genetic diagnostics in Danish autosomal dominant polycystic kidney disease patients reveal possible founder variants.
Local cAMP signalling cascades - roles and targets in chronic kidney disease?
Conserved role of ATP synthase in mammalian cilia.
Rab8 attenuates Wnt signaling and is required for mesenchymal differentiation into adipocytes.
Ccdc113/Ccdc96 complex, a novel regulator of ciliary beating that connects radial spoke 3 to dynein g and the nexin link.
Yes-Associated Protein 1 Is a Novel Calcium Sensing Receptor Target in Human Parathyroid Tumors.
RhoA/Rock activation represents a new mechanism for inactivating Wnt/β-catenin signaling in the aging-associated bone loss.
Expression of α-Tubulin Acetyltransferase 1 and Tubulin Acetylation as Selective Forces in Cell Competition.
Ciliary neuropeptidergic signaling dynamically regulates excitatory synapses in postnatal neocortical pyramidal neurons.
Pathogenic LRRK2 regulates ciliation probability upstream of tau tubulin kinase 2 via Rab10 and RILPL1 proteins.
Transcriptome analysis of ciliary-dependent MCH signaling in differentiating 3T3-L1 pre-adipocytes.
Polyvisceral polycystic disease: a case study and review.
Method for Measuring Mucociliary Clearance and Cilia-generated Flow in Mice by ex vivo Imaging.
On Broken Ne(c)ks and Broken DNA: The Role of Human NEKs in the DNA Damage Response.
CiliOPD: a ciliopathy-associated COPD endotype.
Adherent cell remodeling on micropatterns is modulated by Piezo1 channels.

Eur J Med Genet. 2021 Feb 24. pii: S1769-7212(21)00049-5. [Epub ahead of print] 104183

Clinical genetic diagnostics in Danish autosomal dominant polycystic kidney disease patients reveal possible founder variants.

Marlene L Nielsen, Dorte L Lildballe, Maria Rasmussen, Anders Bojesen, Henrik Birn, Lone Sunde.

  BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is the most common heritable kidney disease. ADPKD leads to cysts, kidney enlargement and end-stage renal disease. ADPKD is mainly caused by variants in PKD1 and PKD2, with truncating PKD1 variants causing the most severe phenotype. This study aimed to characterize variants in Danish patients referred for screening of genes related to cystic kidney disease.METHODS: 147 families were analysed for variants in PKD1, PKD2 and GANAB using next generation sequencing and multiplex ligation-dependent probe amplification. If a variant was identified, relatives were analysed for the specific variant using Sanger sequencing.
RESULTS: A pathogenic or possibly pathogenic variant was identified in 87% (103/118) of patients suspected to suffer from ADPKD, according to the requisition form. In total, 112 pathogenic or possibly pathogenic variants were observed, of which 94 were unique; 74 (79%) in PKD1 and 20 (21%) in PKD2, while 41 variants were novel. No variants in GANAB were observed. Ten recurrent variants were observed in 27 (26%) families. These were either PKD2 variants (N=6) or non-truncating PKD1 variants (N=4). Five of these were likely founder variants.
CONCLUSIONS: The distribution of pathogenic or possibly pathogenic variants in the Danish ADPKD population is similar to that in other populations, except that recurrent truncating PKD1 variants appear to be rare, i.e. founder variants tend to be variant types associated with a mild phenotype. Patients with a mild phenotype may remain undiagnosed, consequently the frequency of founder variants and prevalence of ADPKD may be underestimated.

Keywords:  ADPKD; founder variants; recurrent variants; targeted NGS; variant classification

DOI:  https://doi.org/10.1016/j.ejmg.2021.104183
Acta Physiol (Oxf). 2021 Mar 04. e13641

Local cAMP signalling cascades - roles and targets in chronic kidney disease?

Anastasiia Sholokh, Enno Klussmann.

  The molecular mechanisms underlying chronic kidney disease (CKD) are poorly understood and treatment options are limited, a situation underpinning the need for elucidating the causative molecular mechanisms and for identifying innovative treatment options. It is emerging that cyclic 3',5'-adenosine monophosphate (cAMP) signalling occurs in defined cellular compartments within nanometer dimensions in processes whose dysregulation is associated with CKD. cAMP compartmentalisation is tightly controlled by a specific set of proteins, including A-kinase anchoring proteins (AKAPs) and phosphodiesterases (PDEs). AKAPs such as AKAP18, AKAP220, AKAP-Lbc and STUB1, and PDE4 coordinate arginine-vasopressin (AVP)-induced water reabsorption by collecting duct principal cells. However, hyperactivation of the AVP system is associated with kidney damage and CKD. Podocyte injury involves aberrant AKAP signalling. cAMP signalling in immune cells can be local and slow the progression of inflammatory processes typical for CKD. A major risk factor of CKD is hypertension. cAMP directs the release of the blood pressure regulator, renin, from juxtaglomerular cells, and plays a role in Na+ reabsorption through ENaC, NKCC2 and NCC in the kidney. Mutations in the cAMP hydrolysing PDE3A that cause lowering of cAMP lead to hypertension. Another major risk factor of CKD is diabetes mellitus. AKAP18 and AKAP150 and several PDEs are involved in insulin release. Despite the increasing amount of data, an understanding of functions of compartmentalised cAMP signalling with relevance for CKD is fragmentary. Uncovering functions will improve the understanding of physiological processes and identification of disease-relevant aberrations may guide towards new therapeutic concepts for the treatment of CKD.

Keywords:  A-kinase anchoring protein; ADPKD; AKAP; AQP2; AVP; CKD; PDE; PKA; PKD; PPI; aquaporin-2; arginine-vasopressin; autosomal dominant polycystic kidney disease; cAMP signalling; diabetes; hypertension; phosphodiesterase; polycystic kidney disease; protein kinase A; protein-protein interaction

DOI:  https://doi.org/10.1111/apha.13641
Exp Cell Res. 2021 Feb 24. pii: S0014-4827(21)00051-3. [Epub ahead of print]401(1): 112520

Conserved role of ATP synthase in mammalian cilia.

Wenjun Lin, Cheng Qiao, Jinghua Hu, Qing Wei, Tao Xu.

  We previously found that ATP synthases localize to male-specific sensory cilia and control the ciliary response by regulating polycystin signalling in Caenorhabditis elegans. Herein, we discovered that the ciliary localization of ATP synthase is evolutionarily conserved in mammals. We showed that the ATP synthase subunit F1β is colocalized with the cilia marker acetylated α-tubulin in both mammalian renal epithelial cells (MDCK) and normal mouse cholangiocytes (NMCs). Treatment with ATP synthase inhibitor oligomycin impaired ciliogenesis in MDCK cells, and F1β was co-immunoprecipitated with PKD2 in mammalian cells. Our study provides evidence for the evolutionarily conserved localization of ATP synthase in cilia from worm to mammals. Defects in ATP synthase can lead to ciliary dysfunction, which may be a potential mechanism of polycystic kidney disease.

Keywords:  ATP synthase; Cilia; Polycystin

DOI:  https://doi.org/10.1016/j.yexcr.2021.112520
J Biol Chem. 2021 Mar 01. pii: S0021-9258(21)00262-3. [Epub ahead of print] 100488

Rab8 attenuates Wnt signaling and is required for mesenchymal differentiation into adipocytes.

Ewa Stypulkowski, Qiang Feng, Ivor Joseph, Victoria Farrell, Juan Flores, Shiyan Yu, Ryotaro Sakamori, Jiaxin Sun, Sheila Bandyopadhyay, Soumyashree Das, Radek Dobrowolski, Edward M Bonder, Miao-Hsueh Chen, Nan Gao.

  Differentiation of mesenchymal stem cells into adipocyte requires coordination of external stimuli and depends upon the functionality of primary cilium. The Rab8 small-GTPases are regulators of intracellular transport of membrane-bound structural and signaling cargo. However, the physiological contribution of the intrinsic trafficking network controlled by Rab8 to mesenchymal tissue differentiation has not been fully defined in vivo and in primary tissue cultures. Here, we show that mouse embryonic fibroblasts (MEFs) lacking Rab8 have severely impaired adipocyte differentiation in vivo and ex vivo. Immunofluorescent localization and biochemical analyses of Rab8a-deficient, Rab8b-deficient, and Rab8a and Rab8b double deficient MEFs revealed that Rab8 controls the Lrp6 vesicular compartment, clearance of basal signalosome, traffic of Frizzled 2 receptor, and thereby a proper attenuation of Wnt signaling in differentiating MEFs. Upon induction of adipogenesis program, Rab8a- and Rab8b-deficient MEFs exhibited severely defective lipid droplet formation and abnormal cilia morphology, despite overall intact cilia growth and ciliary cargo transport. Our results suggest that intracellular Rab8 traffic regulates induction of adipogenesis via proper positioning of Wnt receptors for signaling control in mesenchymal cells.

Keywords:  Frizzled; MEF; Rab8; Wnt; adipocyte; differentiation; mouse embryonic fibroblast; primary cilia

DOI:  https://doi.org/10.1016/j.jbc.2021.100488
PLoS Genet. 2021 Mar 04. 17(3): e1009388

Ccdc113/Ccdc96 complex, a novel regulator of ciliary beating that connects radial spoke 3 to dynein g and the nexin link.

Rafał Bazan, Adam Schröfel, Ewa Joachimiak, Martyna Poprzeczko, Gaia Pigino, Dorota Wloga.

Ciliary beating requires the coordinated activity of numerous axonemal complexes. The protein composition and role of radial spokes (RS), nexin links (N-DRC) and dyneins (ODAs and IDAs) is well established. However, how information is transmitted from the central apparatus to the RS and across other ciliary structures remains unclear. Here, we identify a complex comprising the evolutionarily conserved proteins Ccdc96 and Ccdc113, positioned parallel to N-DRC and forming a connection between RS3, dynein g, and N-DRC. Although Ccdc96 and Ccdc113 can be transported to cilia independently, their stable docking and function requires the presence of both proteins. Deletion of either CCDC113 or CCDC96 alters cilia beating frequency, amplitude and waveform. We propose that the Ccdc113/Ccdc96 complex transmits signals from RS3 and N-DRC to dynein g and thus regulates its activity and the ciliary beat pattern.

DOI: https://doi.org/10.1371/journal.pgen.1009388
Int J Mol Sci. 2021 Feb 18. pii: 2016. [Epub ahead of print]22(4):

Yes-Associated Protein 1 Is a Novel Calcium Sensing Receptor Target in Human Parathyroid Tumors.

Giulia Stefania Tavanti, Chiara Verdelli, Annamaria Morotti, Paola Maroni, Vito Guarnieri, Alfredo Scillitani, Rosamaria Silipigni, Silvana Guerneri, Riccardo Maggiore, Gilberto Mari, Leonardo Vicentini, Paolo Dalino Ciaramella, Valentina Vaira, Sabrina Corbetta.

  The Hippo pathway is involved in human tumorigenesis and tissue repair. Here, we investigated the Hippo coactivator Yes-associated protein 1 (YAP1) and the kinase large tumor suppressor 1/2 (LATS1/2) in tumors of the parathyroid glands, which are almost invariably associated with primary hyperparathyroidism. Compared with normal parathyroid glands, parathyroid adenomas (PAds) and carcinomas show variably but reduced nuclear YAP1 expression. The kinase LATS1/2, which phosphorylates YAP1 thus promoting its degradation, was also variably reduced in PAds. Further, YAP1 silencing reduces the expression of the key parathyroid oncosuppressor multiple endocrine neoplasia type 1(MEN1), while MEN1 silencing increases YAP1 expression. Treatment of patient-derived PAds-primary cell cultures and Human embryonic kidney 293A (HEK293A) cells expressing the calcium-sensing receptor (CASR) with the CASR agonist R568 induces YAP1 nuclear accumulation. This effect was prevented by the incubation of the cells with RhoA/Rho-associated coiled-coil-containing protein kinase (ROCK) inhibitors Y27632 and H1152. Lastly, CASR activation increased the expression of the YAP1 gene targets CYR61, CTGF, and WNT5A, and this effect was blunted by YAP1 silencing. Concluding, here we provide preliminary evidence of the involvement of the Hippo pathway in human tumor parathyroid cells and of the existence of a CASR-ROCK-YAP1 axis. We propose a tumor suppressor role for YAP1 and LATS1/2 in parathyroid tumors.

Keywords:  CASR; LATS1/2; MEN1; YAP1; parathormone; parathyroid tumors

DOI:  https://doi.org/10.3390/ijms22042016
Cell Regen. 2021 Mar 03. 10(1): 8

RhoA/Rock activation represents a new mechanism for inactivating Wnt/β-catenin signaling in the aging-associated bone loss.

Wei Shi, Chengyun Xu, Ying Gong, Jirong Wang, Qianlei Ren, Ziyi Yan, Liu Mei, Chao Tang, Xing Ji, Xinhua Hu, Meiyu Qv, Musaddique Hussain, Ling-Hui Zeng, Ximei Wu.

  The Wnt/β-catenin signaling pathway appears to be particularly important for bone homeostasis, whereas nuclear accumulation of β-catenin requires the activation of Rac1, a member of the Rho small GTPase family. The aim of the present study was to investigate the role of RhoA/Rho kinase (Rock)-mediated Wnt/β-catenin signaling in the regulation of aging-associated bone loss. We find that Lrp5/6-dependent and Lrp5/6-independent RhoA/Rock activation by Wnt3a activates Jak1/2 to directly phosphorylate Gsk3β at Tyr216, resulting in Gsk3β activation and subsequent β-catenin destabilization. In line with these molecular events, RhoA loss- or gain-of-function in mouse embryonic limb bud ectoderms interacts genetically with Dkk1 gain-of-function to rescue the severe limb truncation phenotypes or to phenocopy the deletion of β-catenin, respectively. Likewise, RhoA loss-of-function in pre-osteoblasts robustly increases bone formation while gain-of-function decreases it. Importantly, high RhoA/Rock activity closely correlates with Jak and Gsk3β activities but inversely correlates with β-catenin signaling activity in bone marrow mesenchymal stromal cells from elderly male humans and mice, whereas systemic inhibition of Rock therefore activates the β-catenin signaling to antagonize aging-associated bone loss. Taken together, these results identify RhoA/Rock-dependent Gsk3β activation and subsequent β-catenin destabilization as a hitherto uncharacterized mechanism controlling limb outgrowth and bone homeostasis.

Keywords:  Bone; Limb bud; RhoA; Rock; Wnt; β-Catenin

DOI:  https://doi.org/10.1186/s13619-020-00071-3
Cells. 2021 Feb 14. pii: 390. [Epub ahead of print]10(2):

Expression of α-Tubulin Acetyltransferase 1 and Tubulin Acetylation as Selective Forces in Cell Competition.

Amir Mohammad Rahimi, Mingfang Cai, Irem Kılıҫ, Zahra Basir Kazerouni, Constanza Tapia Contreras, Sigrid Hoyer-Fender.

  The wound healing response of fibroblasts critically depends on the primary cilium, a sensory organelle protruding into the environment and comprising a stable axonemal structure. A characteristic marker for primary cilia is acetylation of axonemal tubulin. Although formation of primary cilia is under cell cycle control, the environmental cues affecting ciliation are not fully understood. Our purpose was, therefore, to study the impact of culture conditions on cilia formation in NIH3T3 fibroblasts. We quantified ciliation in different NIH3T3 sub-cell lines and culture conditions by immunodetection of primary cilia and counting. Quantitative Western blotting, qRT-PCR, and proliferation assays completed our investigation. We observed large differences between NIH3T3 sub-cell lines in their ability to generate acetylated primary cilia that correlated with cytoplasmic tubulin acetylation. We found no increased activity of the major tubulin deacetylase, HDAC6, but instead reduced expression of the α-tubulin acetyltransferase 1 (Atat1) as being causative. Our observations demonstrate that cells with reduced expression of Atat1 and tubulin acetylation proliferate faster, eventually displacing all other cells in the population. Expression of Atat1 and tubulin acetylation are therefore selective forces in cell competition.

Keywords:  Atat1 expression; cell competition; cultivation conditions; fibroblasts; primary cilia; tubulin acetylation

DOI:  https://doi.org/10.3390/cells10020390
Elife. 2021 Mar 02. pii: e65427. [Epub ahead of print]10

Ciliary neuropeptidergic signaling dynamically regulates excitatory synapses in postnatal neocortical pyramidal neurons.

Lauren Tereshko, Ya Gao, Brian A Cary, Gina G Turrigiano, Piali Sengupta.

  Primary cilia are compartmentalized sensory organelles present on the majority of neurons in the mammalian brain throughout adulthood. Recent evidence suggests that cilia regulate multiple aspects of neuronal development, including the maintenance of neuronal connectivity. However, whether ciliary signals can dynamically modulate postnatal circuit excitability is unknown. Here we show that acute cell-autonomous knockdown of ciliary signaling rapidly strengthens glutamatergic inputs onto cultured rat neocortical pyramidal neurons, and increases spontaneous firing. This increased excitability occurs without changes to passive neuronal properties or intrinsic excitability. Further, the neuropeptide receptor somatostatin receptor 3 (SSTR3) is localized nearly exclusively to excitatory neuron cilia both in vivo and in culture, and pharmacological manipulation of SSTR3 signaling bidirectionally modulates excitatory synaptic inputs onto these neurons. Our results indicate that ciliary neuropeptidergic signaling dynamically modulates excitatory synapses, and suggest that defects in this regulation may underlie a subset of behavioral and cognitive disorders associated with ciliopathies.

Keywords:  cell biology; neuroscience; rat

DOI:  https://doi.org/10.7554/eLife.65427
Proc Natl Acad Sci U S A. 2021 Mar 09. pii: e2005894118. [Epub ahead of print]118(10):

Pathogenic LRRK2 regulates ciliation probability upstream of tau tubulin kinase 2 via Rab10 and RILPL1 proteins.

Yuriko Sobu, Paulina S Wawro, Herschel S Dhekne, Wondwossen M Yeshaw, Suzanne R Pfeffer.

  Mutations that activate LRRK2 protein kinase cause Parkinson's disease. We showed previously that Rab10 phosphorylation by LRRK2 enhances its binding to RILPL1, and together, these proteins block cilia formation in a variety of cell types, including patient derived iPS cells. We have used live-cell fluorescence microscopy to identify, more precisely, the effect of LRRK2 kinase activity on both the formation of cilia triggered by serum starvation and the loss of cilia seen upon serum readdition. LRRK2 activity decreases the overall probability of ciliation without changing the rates of cilia formation in R1441C LRRK2 MEF cells. Cilia loss in these cells is accompanied by ciliary decapitation, and kinase activity does not change the timing or frequency of decapitation or the rate of cilia loss but increases the percent of cilia that are lost upon serum addition. LRRK2 activity, or overexpression of RILPL1 protein, blocks release of CP110 from the mother centriole, a step normally required for early ciliogenesis; LRRK2 blockade of CP110 uncapping requires Rab10 and RILPL1 proteins and is due to failure to recruit TTBK2, a kinase needed for CP110 release. In contrast, deciliation probability does not change in cells lacking Rab10 or RILPL1 and relies on a distinct LRRK2 pathway. These experiments provide critical detail to our understanding of the cellular consequences of pathogenic LRRK2 mutation and indicate that LRRK2 blocks ciliogenesis upstream of TTBK2 and enhances the deciliation process in response to serum addition.

Keywords:  LRRK2 kinase; Parkinson’s disease; Rab GTPase; primary cilia

DOI:  https://doi.org/10.1073/pnas.2005894118
Sci Rep. 2021 Mar 01. 11(1): 4880

Transcriptome analysis of ciliary-dependent MCH signaling in differentiating 3T3-L1 pre-adipocytes.

Laurie B Cook, Henry D Ophardt, Rongkun Shen, Bryan H Pratt, Lucas A Galbier.

An understanding of adipocyte responsiveness to G-protein-coupled receptor-(GPCR) derived signals must take into consideration the role of membrane microenvironments; that individual sub-populations of proteins may vary significantly across different regions of the cell, and that cell differentiation alters those microenvironments. 3T3-L1 pre-adipocytes undergo a dramatic phenotypic transformation during differentiation into adipocytes, requiring the development of a transient primary cilium. We demonstrate that melanin-concentrating hormone (MCH) receptor 1, a GPCR that stimulates appetite, translocates to the transient primary cilium during early 3T3-L1 cell adipogenesis. Furthermore, we used RNA-Seq to investigate whether MCH signaling is influenced by its receptor localization and whether MCH can influence the transcriptome of early adipocyte development. We found that MCH signaling is sensitive to receptor localization to cilia, and this alters the adipogenic transcriptional program. Also, novel MCH signaling pathways in 3T3-L1 cells are identified, including those for circadian rhythm, the inflammatory response, and ciliary biogenesis. The presence of active MCH-signaling pathways in pre-adipocytes and the discovery that these pathways intersect with the early adipogenic program, among other newly-identified signaling pathways, suggests that the use of MCH receptor 1 antagonists for clinical interventions may have unintended consequences on adipose tissue development.

DOI: https://doi.org/10.1038/s41598-021-84138-4
CEN Case Rep. 2021 Mar 04.

Polyvisceral polycystic disease: a case study and review.

K Indumathi, G Bhavani, K Sudha, G Srinivasaraman, R Manjunathan.

  Polycystic kidney disease (PKD) occurs in one per 20,000 births. Presence of cysts in other organs like adrenal, liver and bladder is even rarer. On reviewing the literature, there is evidence of PKD occurring in conjunction with polycystic liver disease but cysts in multiple viscera are, so far, not reported. A fetal autopsy of a 36-week fetus showed the presence of multiple cysts in the kidney, liver, adrenal and bladder. Further histopathology reports confirmed the diagnosis of polycystic kidney disease. The history of a previous intrauterine death, of another child at 28-week gestation, suggests the presence of familial type. Serial prenatal ultrasonogram did not detect the abnormalities, emphasizing the important role of fetal autopsy in a case with an incomplete obstetric history. The diagnosis of a fetal abnormality aids to counselling the parents to be aware of possible recurrences in new pregnancies.

Keywords:  Autopsy; Chromosomal microarray; Polycystic kidney disease

DOI:  https://doi.org/10.1007/s13730-021-00582-1
Bio Protoc. 2020 Mar 20. 10(6): e3554

Method for Measuring Mucociliary Clearance and Cilia-generated Flow in Mice by ex vivo Imaging.

Eric Song, Akiko Iwasaki.

  Ex vivo biophysical measurements provide valuable insights into understanding both physiological and pathogenic processes. One critical physiological mechanism that is regulated by these biophysical properties is cilia-generated flow that mediates mucociliary clearance, which is known to provide protection against foreign particles and pathogens in the upper airway. To measure ciliary clearance, several techniques have been implemented, including the use of radiolabeled particles and imaging with single-photon emission computerized tomography (SPECT) methods. Although non-invasive, these tests require the use of specialized equipment, limiting widespread use. Here we describe a method of ex vivo imaging of cilia-generated flow, adapted from previously reported methods, to make it more accessible and higher throughput for researchers. We excise trachea from mice quickly after euthanasia, cut it longitudinally and place it in an inhouse made slide. We apply fluorescent particles to measure particle movement under a fluorescent microscope, followed by analysis with ImageJ, allowing calculation of fluid flow generated by cilia under different conditions. This method enables ex vivo measurements in tissue with minimal investment or special equipment, giving opportunity to investigate and discover important biophysical properties associated with ciliary movement of the trachea in physiology and disease.

Keywords:  Biophysics; Cilia; Ex-vivo imaging; Lung; Mucociliary clearance

DOI:  https://doi.org/10.21769/BioProtoc.3554
Cells. 2021 Feb 27. pii: 507. [Epub ahead of print]10(3):

On Broken Ne(c)ks and Broken DNA: The Role of Human NEKs in the DNA Damage Response.

Isadora Carolina Betim Pavan, Andressa Peres de Oliveira, Pedro Rafael Firmino Dias, Fernanda Luisa Basei, Luidy Kazuo Issayama, Camila de Castro Ferezin, Fernando Riback Silva, Ana Luisa Rodrigues de Oliveira, Lívia Alves Dos Reis Moura, Mariana Bonjiorno Martins, Fernando Moreira Simabuco, Jörg Kobarg.

  NIMA-related kinases, or NEKs, are a family of Ser/Thr protein kinases involved in cell cycle and mitosis, centrosome disjunction, primary cilia functions, and DNA damage responses among other biological functional contexts in vertebrate cells. In human cells, there are 11 members, termed NEK1 to 11, and the research has mainly focused on exploring the more predominant roles of NEKs in mitosis regulation and cell cycle. A possible important role of NEKs in DNA damage response (DDR) first emerged for NEK1, but recent studies for most NEKs showed participation in DDR. A detailed analysis of the protein interactions, phosphorylation events, and studies of functional aspects of NEKs from the literature led us to propose a more general role of NEKs in DDR. In this review, we express that NEK1 is an activator of ataxia telangiectasia and Rad3-related (ATR), and its activation results in cell cycle arrest, guaranteeing DNA repair while activating specific repair pathways such as homology repair (HR) and DNA double-strand break (DSB) repair. For NEK2, 6, 8, 9, and 11, we found a role downstream of ATR and ataxia telangiectasia mutated (ATM) that results in cell cycle arrest, but details of possible activated repair pathways are still being investigated. NEK4 shows a connection to the regulation of the nonhomologous end-joining (NHEJ) repair of DNA DSBs, through recruitment of DNA-PK to DNA damage foci. NEK5 interacts with topoisomerase IIβ, and its knockdown results in the accumulation of damaged DNA. NEK7 has a regulatory role in the detection of oxidative damage to telomeric DNA. Finally, NEK10 has recently been shown to phosphorylate p53 at Y327, promoting cell cycle arrest after exposure to DNA damaging agents. In summary, this review highlights important discoveries of the ever-growing involvement of NEK kinases in the DDR pathways. A better understanding of these roles may open new diagnostic possibilities or pharmaceutical interventions regarding the chemo-sensitizing inhibition of NEKs in various forms of cancer and other diseases.

Keywords:  DNA damage response; cell cycle; kinase; protein kinase

DOI:  https://doi.org/10.3390/cells10030507
Respir Res. 2021 Feb 27. 22(1): 74

CiliOPD: a ciliopathy-associated COPD endotype.

Jeanne-Marie Perotin, Myriam Polette, Gaëtan Deslée, Valérian Dormoy.

  The pathophysiology of chronic obstructive pulmonary disease (COPD) relies on airway remodelling and inflammation. Alterations of mucociliary clearance are a major hallmark of COPD caused by structural and functional cilia abnormalities. Using transcriptomic databases of whole lung tissues and isolated small airway epithelial cells (SAEC), we comparatively analysed cilia-associated and ciliopathy-associated gene signatures from a set of 495 genes in 7 datasets including 538 non-COPD and 508 COPD patients. This bio-informatics approach unveils yet undescribed cilia and ciliopathy genes associated with COPD including NEK6 and PROM2 that may contribute to the pathology, and suggests a COPD endotype exhibiting ciliopathy features (CiliOPD).

Keywords:  COPD; Cilia; Transcriptomic

DOI:  https://doi.org/10.1186/s12931-021-01665-4
Sci Rep. 2021 Mar 03. 11(1): 5088

Adherent cell remodeling on micropatterns is modulated by Piezo1 channels.

Deekshitha Jetta, Mohammad Reza Bahrani Fard, Frederick Sachs, Katie Munechika, Susan Z Hua.

Adherent cells utilize local environmental cues to make decisions on their growth and movement. We have previously shown that HEK293 cells grown on the fibronectin stripe patterns were elongated. Here we show that Piezo1 function is involved in cell spreading. Piezo1 expressing HEK cells plated on fibronectin stripes elongated, while a knockout of Piezo1 eliminated elongation. Inhibiting Piezo1 conductance using GsMTx4 or Gd3+ blocked cell spreading, but the cells grew thin tail-like extensions along the patterns. Images of GFP-tagged Piezo1 showed plaques of Piezo1 moving to the extrusion edges, co-localized with focal adhesions. Surprisingly, in non-spreading cells Piezo1 was located primarily on the nuclear envelope. Inhibiting the Rho-ROCK pathway also reversibly inhibited cell extension indicating that myosin contractility is involved. The growth of thin extrusion tails did not occur in Piezo1 knockout cells suggesting that Piezo1 may have functions besides acting as a cation channel.

DOI: https://doi.org/10.1038/s41598-021-84427-y