Inhibiting Focal Adhesion Kinase Ameliorates Cyst Development in Polycystin-1–Deficient Polycystic Kidney Disease in Animal Model

2021 ◽  
Vol 32 (9) ◽  
pp. 2159-2174
Author(s):  
Jinzhao He ◽  
Shun Zhang ◽  
Zhiwei Qiu ◽  
Xiaowei Li ◽  
Huihui Huang ◽  
...  
Keyword(s):  
Mouse Models ◽  
Focal Adhesion ◽  
Knockout Mouse ◽  
Ex Vivo ◽  
Renal Cyst ◽  
Tubular Epithelial Cells ◽  
Polycystic Kidney ◽  
Content Type ◽  
Cyst Development

BackgroundAutosomal dominant polycystic kidney disease (ADPKD) is characterized by numerous cysts originating from renal tubules and is associated with significant tubular epithelial cell proliferation. Focal adhesion kinase (FAK) promotes tumor growth by regulating multiple proliferative pathways.MethodsWe established the forskolin (FSK)-induced three-dimensional (3D) Madin–Darby Canine Kidney cystogenesis model and 8-bromoadenosine-3`,5`-cyclic monophosphate–stimulated cyst formation in ex vivo embryonic kidney culture. Cultured human renal cyst–lining cells (OX-161) and normal tubular epithelial cells were treated with FAK inhibitors or transfected with green fluorescent protein–tagged FAK mutant plasmids for proliferation study. Furthermore, we examined the role of FAK in two transgenic ADPKD animal models, the kidney-specific Pkd1 knockout and the collecting duct–specific Pkd1 knockout mouse models.ResultsFAK activity was significantly elevated in OX-161 cells and in two ADPKD mouse models. Inhibiting FAK activity reduced cell proliferation in OX-161 cells and prevented cyst growth in ex vivo and 3D cyst models. In tissue-specific Pkd1 knockout mouse models, FAK inhibitors retarded cyst development and mitigated renal function decline. Mechanically, FSK stimulated FAK activation in tubular epithelial cells, which was blocked by a protein kinase A (PKA) inhibitor. Inhibition of FAK activation by inhibitors or transfected cells with mutant FAK constructs interrupted FSK-mediated Src activation and upregulation of ERK and mTOR pathways.ConclusionsOur study demonstrates the critical involvement of FAK in renal cyst development, suggests that FAK is a potential therapeutic target in treating patients with ADPKD, and highlights the role of FAK in cAMP-PKA–regulated proliferation.

scholarly journals Obacunone Retards Renal Cyst Development in Autosomal Dominant Polycystic Kidney Disease by Activating NRF2

Antioxidants ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 38
Author(s):  
Zhiwei Qiu ◽  
Jinzhao He ◽  
Guangying Shao ◽  
Jiaqi Hu ◽  
Xiaowei Li ◽  
...  
Keyword(s):  
Kidney Disease ◽  
Polycystic Kidney Disease ◽  
Autosomal Dominant ◽  
Renal Cyst ◽  
Renal Diseases ◽  
Therapeutic Drug ◽  
Dependent Manner ◽  
Polycystic Kidney ◽  
Autosomal Dominant Polycystic Kidney ◽  
Cyst Development

Autosomal dominant polycystic kidney disease (ADPKD) is a common inherited disease characterized by progressive enlargement of fluid-filled cysts derived from renal tubular epithelial cells, which has become the fourth leading cause of end-stage renal diseases. Currently, treatment options for ADPKD remain limited. The purpose of this study was to discover an effective therapeutic drug for ADPKD. With virtual screening, Madin-Darby canine kidney (MDCK) cyst model, embryonic kidney cyst model and kidney-specific Pkd1 knockout mouse (PKD) model, we identified obacunone as a candidate compound for ADPKD drug discovery from a natural antioxidant compound library. In vitro experiments showed that obacunone significantly inhibited cyst formation and expansion of MDCK cysts and embryonic kidney cysts in a dose-dependent manner. In vivo, obacunone treatment significantly reduced the renal cyst development in PKD mice. Western blot and morphological analysis revealed that obacunone served as a NRF2 activator in ADPKD, which suppressed lipid peroxidation by up-regulating GPX4 and finally restrained excessive cell proliferation by down-regulating mTOR and MAPK signaling pathways. Experimental data demonstrated obacunone as an effective renal cyst inhibitor for ADPKD, indicating that obacunone might be developed into a therapeutic drug for ADPKD treatment.

scholarly journals Influence of CYP2C8, CYP3A4, and CYP3A5 Host Genotypes on Early Recurrence of Plasmodium vivax

2020 ◽  
Vol 64 (7) ◽  
Author(s):  
Anne C. G. Almeida ◽  
Maria C. B. Puça ◽  
Erick F. G. Figueiredo ◽  
Laila R. Barbosa ◽  
Yanka E. A. R. Salazar ◽  
...  
Keyword(s):  
Plasmodium Vivax ◽  
Ex Vivo ◽  
Plasma Concentrations ◽  
Similar Efficacy ◽  
Early Recurrence ◽  
Control Group ◽  
Recurrence Rates ◽  
Mutant Genotype ◽  
Content Type

ABSTRACT Cytochrome P450 (CYP) enzymes are involved in the biotransformation of chloroquine (CQ), but the role of the different profiles of metabolism of this drug in relation to Plasmodium vivax recurrences has not been properly investigated. To investigate the influence of the CYP genotypes associated with CQ metabolism on the rates of P. vivax early recurrences, a case-control study was carried out. The cases included patients presenting with an early recurrence (CQ-recurrent individuals), defined as a recurrence during the first 28 days after initial infection and plasma concentrations of CQ plus desethylchloroquine (DCQ; the major CQ metabolite) higher than 100 ng/ml. A control group with no parasite recurrence over the follow-up (the CQ-responsive group) was also included. CQ and DCQ plasma levels were measured on day 28. CQ-metabolizing CYP (CYP2C8, CYP3A4, and CYP3A5) genotypes were determined by real-time PCR. An ex vivo study was conducted to verify the efficacy of CQ and DCQ against P. vivax isolates. The frequency of alleles associated with normal and slow metabolism was similar between the cases and the controls for the CYP2C8 (odds ratio [OR] = 1.45, 95% confidence interval [CI] = 0.51 to 4.14, P = 0.570), CYP3A4 (OR = 2.38, 95% CI = 0.92 to 6.19, P = 0.105), and CYP3A5 (OR = 4.17, 95% CI = 0.79 to 22.04, P = 1.038) genes. DCQ levels were higher than CQ levels, regardless of the genotype. Regarding the DCQ/CQ ratio, there was no difference between groups or between those patients who had a normal genotype and those patients who had a mutant genotype. DCQ and CQ showed similar efficacy ex vivo. CYP genotypes had no influence on early recurrence rates. The similar efficacy of CQ and DCQ ex vivo could explain the absence of therapeutic failure, despite the presence of alleles associated with slow metabolism.

scholarly journals Na+/H+Antiport Is Essential for Yersinia pestis Virulence

2013 ◽  
Vol 81 (9) ◽  
pp. 3163-3172 ◽  
Author(s):  
Yusuke Minato ◽  
Amit Ghosh ◽  
Wyatt J. Faulkner ◽  
Erin J. Lind ◽  
Sara Schesser Bartra ◽  
...  
Keyword(s):  
Yersinia Pestis ◽  
Drug Target ◽  
Deletion Mutant ◽  
Ex Vivo ◽  
Ion Homeostasis ◽  
Content Type ◽  
Double Deletion ◽  
Derivative Strain

ABSTRACTNa+/H+antiporters are ubiquitous membrane proteins that play a central role in the ion homeostasis of cells. In this study, we examined the possible role of Na+/H+antiport inYersinia pestisvirulence and found thatY. pestisstrains lacking the major Na+/H+antiporters, NhaA and NhaB, are completely attenuated in anin vivomodel of plague. TheY. pestisderivative strain lacking thenhaAandnhaBgenes showed markedly decreased survival in blood and blood serumex vivo. Complementation of eithernhaAornhaBintransrestored the survival of theY. pestis nhaA nhaBdouble deletion mutant in blood. ThenhaA nhaBdouble deletion mutant also showed inhibited growth in an artificial serum medium, Opti-MEM, and a rich LB-based medium with Na+levels and pH values similar to those for blood. Taken together, these data strongly suggest that intact Na+/H+antiport is indispensable for the survival ofY. pestisin the bloodstreams of infected animals and thus might be regarded as a promising noncanonical drug target for infections caused byY. pestisand possibly for those caused by other blood-borne bacterial pathogens.

scholarly journals Dectin-1-Dependent Interleukin-22 Contributes to Early Innate Lung Defense against Aspergillus fumigatus

2011 ◽  
Vol 80 (1) ◽  
pp. 410-417 ◽  
Author(s):  
Melissa A. Gessner ◽  
Jessica L. Werner ◽  
Lauren M. Lilly ◽  
Michael P. Nelson ◽  
Allison E. Metz ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Ex Vivo ◽  
Lung Cells ◽  
Lipocalin 2 ◽  
Beta Glucan ◽  
Necrosis Factor Alpha ◽  
Content Type ◽  
Deficient Mice ◽  
Lung Defense

ABSTRACTWe have previously reported that mice deficient in the beta-glucan receptor Dectin-1 displayed increased susceptibility toAspergillus fumigatuslung infection in the presence of lower interleukin 23 (IL-23) and IL-17A production in the lungs and have reported a role for IL-17A in lung defense. As IL-23 is also thought to control the production of IL-22, we examined the role of Dectin-1 in IL-22 production, as well as the role of IL-22 in innate host defense againstA. fumigatus. Here, we show that Dectin-1-deficient mice demonstrated significantly reduced levels of IL-22 in the lungs early afterA. fumigatuschallenge. Culturing cells from enzymatic lung digestsex vivofurther demonstrated Dectin-1-dependent IL-22 production. IL-22 production was additionally found to be independent of IL-1β, IL-6, or IL-18 but required IL-23. The addition of recombinant IL-23 augmented IL-22 production in wild-type (WT) lung cells and rescued IL-22 production by lung cells from Dectin-1-deficient mice.In vivoneutralization of IL-22 in the lungs of WT mice resulted in impairedA. fumigatuslung clearance. Moreover, mice deficient in IL-22 also demonstrated a higher lung fungal burden afterA. fumigatuschallenge in the presence of impaired IL-1α, tumor necrosis factor alpha (TNF-α), CCL3/MIP-1α, and CCL4/MIP-1β production and lower neutrophil recruitment, yet intact IL-17A production. We further show that lung lavage fluid collected from bothA. fumigatus-challenged Dectin-1-deficient and IL-22-deficient mice had compromised anti-fungal activity againstA. fumigatus in vitro. Although lipocalin 2 production was observed to be Dectin-1 and IL-22 dependent, lipocalin 2-deficient mice did not demonstrate impairedA. fumigatusclearance. Moreover, lungS100a8,S100a9, andReg3gmRNA expression was not lower in either Dectin-1-deficient or IL-22-deficient mice. Collectively, our results indicate that early innate lung defense againstA. fumigatusis mediated by Dectin-1-dependent IL-22 production.

scholarly journals Contrasting role of NLRP12 in autoinflammation: evidence from a case report and mouse models

RMD Open ◽  
2021 ◽  
Vol 7 (3) ◽  
pp. e001824
Author(s):  
Dan Lévy ◽  
Alexandre Mariotte ◽  
Aurore DeCauwer ◽  
Cecile Macquin ◽  
Angélique Pichot ◽  
...  
Keyword(s):  
Mouse Models ◽  
Ex Vivo ◽  
Joint Inflammation ◽  
Autoinflammatory Syndrome ◽  
Her Family ◽  
Whole Exome ◽  
Urate Crystals

ObjectiveTo explore at the molecular level the phenotype of a patient suffering an autoinflammatory syndrome which was diagnosed as familial cold autoinflammatory syndrome type 2 (FCAS-2). To explore the functions of Nlrp12 in inflammation using mouse models.MethodsWhole exome sequencing and Nlrp12 targeted resequencing were performed on DNA isolated from the patient and her family members. In vivo and ex vivo models of inflammation (urate crystals-dependent acute joint inflammation and urate crystals-induced peritonitis) were analysed in Nlrp12-deficient and Nlrp12-competent mice.ResultsA rare missense NLRP12 variant (c.857C>T, p.P286L) was identified in the patient and her healthy relatives. Nlrp12-deficient mice exhibit reduced systemic inflammation and neutrophilic infiltration.ConclusionNlrp12 mediates proinflammatory functions in mice. In humans, the identification of Nlrp12 variants must be cautiously interpreted depending on clinical and paraclinical data to diagnose FCAS-2.

scholarly journals Is There a Role of ADAMTS-1 in Cyst Development in Autosomal Dominant Polycystic Kidney Disease?

2021 ◽  
Vol 30 (1) ◽  
pp. 25-29
Author(s):  
Suleyman Karakose ◽  
◽  
Pervin Ozkan Kurtgoz ◽  
Cigdem Damla Deniz ◽  
Edip Erkus ◽  
...  
Keyword(s):  
Kidney Disease ◽  
Polycystic Kidney Disease ◽  
Autosomal Dominant ◽  
Polycystic Kidney ◽  
Autosomal Dominant Polycystic Kidney ◽  
Cyst Development

scholarly journals Ganoderic acid A is the effective ingredient of Ganoderma triterpenes in retarding renal cyst development in polycystic kidney disease

2020 ◽  
Vol 41 (6) ◽  
pp. 782-790 ◽  
Author(s):  
Jia Meng ◽  
Sai-zhen Wang ◽  
Jin-zhao He ◽  
Shuai Zhu ◽  
Bo-yue Huang ◽  
...  
Keyword(s):  
Kidney Disease ◽  
Polycystic Kidney Disease ◽  
Renal Cyst ◽  
Polycystic Kidney ◽  
Ganoderic Acid ◽  
Cyst Development

scholarly journals Intestinally Secreted C-Type Lectin Reg3b Attenuates Salmonellosis but Not Listeriosis in Mice

2012 ◽  
Vol 80 (3) ◽  
pp. 1115-1120 ◽  
Author(s):  
Marleen T. J. van Ampting ◽  
Linda M. P. Loonen ◽  
Arjan J. Schonewille ◽  
Irene Konings ◽  
Carolien Vink ◽  
...  
Keyword(s):  
Bacterial Infections ◽  
Salmonella Enteritidis ◽  
Ex Vivo ◽  
Protective Role ◽  
Carbohydrate Binding ◽  
Intestinal Infection ◽  
Gram Positive ◽  
Gram Negative ◽  
Content Type

The Reg3 protein family, including the human member designated pancreatitis-associated protein (PAP), consists of secreted proteins that contain a C-type lectin domain involved in carbohydrate binding. They are expressed by intestinal epithelial cells. Colonization of germ-free mice and intestinal infection with pathogens increase the expression of Reg3g and Reg3b in the murine ileum. Reg3g is directly bactericidal for Gram-positive bacteria, but the exact role of Reg3b in bacterial infections is unknown. To investigate the possible protective role of Reg3b in intestinal infection, Reg3b knockout (Reg3b−/−) mice and wild-type (WT) mice were orally infected with Gram-negativeSalmonella enteritidisor Gram-positiveListeria monocytogenes. At day 2 after oralListeriainfection and at day 4 after oralSalmonellainfection, mice were sacrificed to collect intestinal and other tissues for pathogen quantification. Protein expression of Reg3b and Reg3g was determined in intestinal mucosal scrapings of infected and noninfected mice. In addition,ex vivobinding of ileal mucosal Reg3b toListeriaandSalmonellawas investigated. Whereas recovery ofSalmonellaorListeriafrom feces of Reg3b−/−mice did not differ from that from feces of WT mice, significantly higher numbers of viableSalmonella, but notListeria, bacteria were recovered from the colon, mesenteric lymph nodes, spleen, and liver of the Reg3b−/−mice than from those of WT mice. Mucosal Reg3b binds to both bacterial pathogens and may interfere with their mode of action. Reg3b plays a protective role against intestinal translocation of the Gram-negative bacteriumS. enteritidisin mice but not against the Gram-positive bacteriumL. monocytogenes.

scholarly journals Role of Dendritic Cells in the Pathogenesis of Whipple's Disease

2014 ◽  
Vol 83 (2) ◽  
pp. 482-491 ◽  
Author(s):  
Katina Schinnerling ◽  
Anika Geelhaar-Karsch ◽  
Kristina Allers ◽  
Julian Friebel ◽  
Kristina Conrad ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Interleukin 12 ◽  
Lymphoid Tissues ◽  
Whipple’S Disease ◽  
Whipple's Disease ◽  
Content Type ◽  
Healthy Donors ◽  
Stimulatory Capacity

Accumulation ofTropheryma whipplei-stuffed macrophages in the duodenum, impairedT. whipplei-specific Th1 responses, and weak secretion of interleukin-12 (IL-12) are hallmarks of classical Whipple's disease (CWD). This study addresses dendritic cell (DC) functionality during CWD. We documented composition, distribution, and functionality of DCex vivoor afterin vitromaturation by fluorescence-activated cell sorting (FACS) and by immunohistochemistryin situ. A decrease in peripheral DC of untreated CWD patients compared to healthy donors was due to reduced CD11chighmyeloid DC (M-DC). Decreased maturation markers CD83, CD86, and CCR7, as well as low IL-12 production in response to stimulation, disclosed an immature M-DC phenotype.In vitro-generated monocyte-derived DC from CWD patients showed normal maturation and T cell-stimulatory capacity under proinflammatory conditions but produced less IL-12 and failed to activateT. whipplei-specific Th1 cells. In duodenal and lymphoid tissues,T. whippleiwas found within immature DC-SIGN+DC. DC and proliferating lymphocytes were reduced in lymph nodes of CWD patients compared to levels in controls. Our results indicate that dysfunctional IL-12 production by DC provides suboptimal conditions for priming ofT. whipplei-specific T cells during CWD and that immature DC carryingT. whippleicontribute to the dissemination of the bacterium.

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