Experimental Inhibition of Periostin Attenuates Kidney Fibrosis

2017 ◽  
Vol 46 (6) ◽  
pp. 501-517 ◽  
Author(s):  
Jin Ho Hwang ◽  
Seung Hee Yang ◽  
Yong Chul Kim ◽  
Jin Hyuk Kim ◽  
Jung Nam An ◽  
...  
Keyword(s):  
Renal Fibrosis ◽  
Matrix Protein ◽  
Collecting Duct ◽  
Extracellular Matrix Protein ◽  
In Vitro Experiments ◽  
Wild Type ◽  
In The Wild ◽  
Renal Fibrogenesis

Background: Periostin is responsible for tissue regeneration, fibrosis, and wound healing via its interaction with integrin. Recently, the role of periostin has been shown to contribute to fibrosis in chronic kidney disease. We investigated the role of periostin and the effect of periostin blockade in renal fibrogenesis. Methods: We investigated the function of periostin in vivo in wild-type and periostin-null mice (Postn-KO) in a unilateral ureteral obstruction (UUO) model. For the in vitro experiments, primary cultured inner medullary collecting duct cells from the wild-type and Postn-KO mice were used. Results: Periostin expression was strongly induced by UUO in the wild-type mice. UUO induced renal fibrosis and morphological changes in the obstructed kidney of wild-type mice, whereas global knockout of periostin reduced fibrosis induced by UUO and improved kidney structure. Fibrosis- and inflammation-related mRNA were significantly induced in the wild-type mice and were decreased in the Postn-KO mice. Additionally, α-smooth muscle actin expression was increased following the administration of recombinant periostin in vitro. The effect of periostin blockade was examined using 2 methods. The integrin blockade peptide decreased fibrosis-related gene expression in in vitro experiments. Anti-periostin polyclonal antibody attenuated renal fibrosis induced by UUO through changes in transforming growth factor-β signaling and the inflammatory and apoptotic pathways. Conclusion: Periostin is a marker of renal fibrosis and may augment the progression of fibrogenesis as an extracellular matrix protein. Periostin blockade effectively attenuated renal fibrogenesis. Thus, periostin inhibition may be a therapeutic strategy for the amelioration of renal disease progression.

scholarly journals Mutant Dentin Sialophosphoprotein Causes Dentinogenesis Imperfecta

2019 ◽  
Vol 98 (8) ◽  
pp. 912-919 ◽  
Author(s):  
T. Liang ◽  
H. Zhang ◽  
Q. Xu ◽  
S. Wang ◽  
C. Qin ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Matrix Protein ◽  
Mutant Mice ◽  
Extracellular Matrix Protein ◽  
Dentinogenesis Imperfecta ◽  
Wild Type ◽  
Pulp Chamber ◽  
Dentin Sialophosphoprotein ◽  
In The Wild

Dentin sialophosphoprotein (DSPP) is an extracellular matrix protein highly expressed by odontoblasts in teeth. DSPP mutations in humans may cause dentinogenesis imperfecta (DGI), an autosomal dominant dentin disorder. We recently generated a mouse model (named “ DsppP19L/+ mice”) that expressed a mutant DSPP in which the proline residue at position 19 was replaced by a leucine residue. We found that the DsppP19L/+ and DsppP19L/P19L mice at a younger age displayed a tooth phenotype resembling human DGI type III characterized by enlarged dental pulp chambers, while the teeth of older DsppP19L/+ and DsppP19L/P19L mice had smaller dental pulp chambers mimicking DGI type II. The teeth of DsppP19L/+ and DsppP19L/P19L mice had a narrower pulp chamber roof predentin layer, thinner pulp chamber roof dentin, and thicker pulp chamber floor dentin. In addition, these mice also had increased enamel attrition, accompanied by excessive deposition of peritubular dentin. Immunohistochemistry, in situ hybridization, and real-time polymerase chain reaction analyses showed that the odontoblasts in both DsppP19L/+ and DsppP19L/P19L mice had reduced DSPP expression, compared to the wild-type mice. We also observed that the levels of DSPP expression were much higher in the roof-forming odontoblasts than in the floor-forming odontoblasts in the wild-type mice and mutant mice. Moreover, immunohistochemistry showed that while the immunostaining signals of dentin sialoprotein (N-terminal fragment of DSPP) were decreased in the dentin matrix, they were remarkably increased in the odontoblasts of the DsppP19L/+ and DsppP19L/P19L mice. Consistently, our in vitro studies showed that the secretion of the mutant DSPP was impaired and accumulated within endoplasmic reticulum. These findings suggest that the dental phenotypes of the mutant mice were associated with the intracellular retention of the mutant DSPP in the odontoblasts of the DSPP-mutant mice.

scholarly journals LTBP1 promotes fibrillin incorporation into the extracellular matrix

2021 ◽  
Author(s):  
Matthias Przyklenk ◽  
Veronika Georgieva ◽  
Fabian Metzen ◽  
Sebastian Mostert ◽  
Birgit Kobbe ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Matrix Protein ◽  
Wide Spectrum ◽  
Extracellular Matrix Protein ◽  
Connective Tissues ◽  
Pathogenic Variants ◽  
Human Pathology ◽  
Fibrillin 1

LTBP1 is a large extracellular matrix protein and an associated ligand of fibrillin-microfibrils. Knowledge of LTBP1 functions is largely limited to its role in targeting and sequestering TGFβ growth factors within the extracellular matrix, thereby regulating their bioavailability. However, the recent description of a wide spectrum of phenotypes in multiple tissues in patients harboring LTBP1 pathogenic variants suggests a multifaceted role of the protein in the homeostasis of connective tissues. To better understand the human pathology caused by LTBP1 deficiency it is important to investigate its functional role in extracellular matrix formation. In this study, we show that LTBP1 coordinates the incorporation of fibrillin-1 and -2 into the extracellular matrix in vitro. We also demonstrate that this function is differentially exerted by the two isoforms, the short and long forms of LTBP1. Thereby our findings uncover a novel TGFβ-independent LTBP1 function potentially contributing to the development of connective tissue disorders.

Role of mesenchymal nidogen for epithelial morphogenesis in vitro

Development ◽  
1994 ◽  
Vol 120 (7) ◽  
pp. 2003-2014 ◽  
Author(s):  
P. Ekblom ◽  
M. Ekblom ◽  
L. Fecker ◽  
G. Klein ◽  
H.Y. Zhang ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Northern Blotting ◽  
Epithelial Morphogenesis ◽  
Extracellular Matrix Protein ◽  
Epithelial Development ◽  
Morphogenesis In Vitro ◽  
Development In Vitro ◽  
Membrane Components

Recent biochemical studies suggested that the extracellular matrix protein nidogen is a binding molecule linking together basement membrane components. We studied its expression and role during development. By immunofluorescence and northern blotting, nidogen was found early during epithelial cell development of kidney and lung. Yet, in situ hybridization revealed that nidogen was not produced by epithelium but by the adjacent mesenchyme in both organs. Binding of mesenchymal nidogen to epithelial laminin may thus be a key event during epithelial development. This is supported by antibody perturbation experiments. Antibodies against the nidogen binding site on laminin B2 chain perturbed epithelial development in vitro in embryonic kidney and lung. Mesenchymal nidogen could be important for early stages of epithelial morphogenesis.

scholarly journals Colonization and Inflammation Deficiencies in Mongolian Gerbils Infected by Helicobacter pylori Chemotaxis Mutants

2005 ◽  
Vol 73 (3) ◽  
pp. 1820-1827 ◽  
Author(s):  
David J. McGee ◽  
Melanie L. Langford ◽  
Emily L. Watson ◽  
J. Elliot Carter ◽  
Yu-Ting Chen ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Immune Cell ◽  
Response Regulator ◽  
Critical Role ◽  
Mongolian Gerbils ◽  
Wild Type ◽  
In The Wild ◽  
H Pylori

ABSTRACT Helicobacter pylori causes disease in the human stomach and in mouse and gerbil stomach models. Previous results have shown that motility is critical for H. pylori to colonize mice, gerbils, and other animal models. The role of chemotaxis, however, in colonization and disease is less well understood. Two genes in the H. pylori chemotaxis pathway, cheY and tlpB, which encode the chemotaxis response regulator and a methyl-accepting chemoreceptor, respectively, were disrupted. The cheY mutation was complemented with a wild-type copy of cheY inserted into the chromosomal rdxA gene. The cheY mutant lost chemotaxis but retained motility, while all other strains were motile and chemotactic in vitro. These strains were inoculated into gerbils either alone or in combination with the wild-type strain, and colonization and inflammation were assessed. While the cheY mutant completely failed to colonize gerbil stomachs, the tlpB mutant colonized at levels similar to those of the wild type. With the tlpB mutant, there was a substantial decrease in inflammation in the gerbil stomach compared to that with the wild type. Furthermore, there were differences in the numbers of each immune cell in the tlpB-mutant-infected stomach: the ratio of lymphocytes to neutrophils was about 8 to 1 in the wild type but only about 1 to 1 in the mutant. These results suggest that the TlpB chemoreceptor plays an important role in the inflammatory response while the CheY chemotaxis regulator plays a critical role in initial colonization. Chemotaxis mutants may provide new insights into the steps involved in H. pylori pathogenesis.

Single-chain antibody fragments derived from a human synthetic phage-display library bind thrombospondin and inhibit sickle cell adhesion

Blood ◽  
2003 ◽  
Vol 102 (2) ◽  
pp. 718-724 ◽  
Author(s):  
Nicholas A. Watkins ◽  
Lily M. Du ◽  
J. Paul Scott ◽  
Willem H. Ouwehand ◽  
Cheryl A. Hillery
Keyword(s):  
Sickle Cell ◽  
Matrix Protein ◽  
Binding Domain ◽  
Extracellular Matrix Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adhesion Assay ◽  
Single Chain ◽  
Cell Binding

AbstractThe enhanced adhesion of sickle red blood cells (RBCs) to the vascular endothelium and subendothelial matrix likely plays a significant role in the pathogenesis of vaso-occlusion in sickle cell disease. Sickle RBCs have enhanced adhesion to the plasma and extracellular matrix protein thrombospondin-1 (TSP) under conditions of flow in vitro. In this study, we sought to develop antibodies that bind TSP from a highly diverse library of human single-chain Fv fragments (scFvs) displayed on filamentous phage. Following 3 rounds of phage selection of increasing stringency 6 unique scFvs that bound purified TSP by enzyme-linked immunosorbent assay were isolated. Using an in vitro flow adhesion assay, 3 of the 6 isolated scFvs inhibited the adhesion of sickle RBCs to immobilized TSP by more than 40% compared with control scFvs (P < .001). Furthermore, scFv TSP-A10 partially inhibited sickle RBC adhesion to activated endothelial cells (P < .005). Using TSP proteolytic fragments to map the binding site, we showed that 2 of the inhibitory scFvs bound an epitope in the calcium-binding domain or proximal cell-binding domain of TSP, providing evidence for the role of these domains in the adhesion of sickle RBCs to TSP. In summary, we have isolated a panel of scFvs that specifically bind to TSP and differentially inhibit sickle RBC adhesion to surface-bound TSP under flow conditions. These scFvs will be useful reagents for investigating the role of the calcium and cell-binding domains of TSP in sickle RBC adhesion.

Abstract 291: Fibulin-1 is Required to Prevent Hypoplastic Semilunar Valve Formation

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Keerthi Harikrishnan ◽  
Marion A Cooley ◽  
Waleed O Twal ◽  
Victor M Fresco ◽  
Christine B Kern ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Pulmonary Valve ◽  
Extracellular Matrix Protein ◽  
Semilunar Valve ◽  
Anterior Leaflet ◽  
Wild Type ◽  
Endocardial Cushions ◽  
Cell Numbers ◽  
Valve Formation

Fibulin-1 (Fbln1), an extracellular matrix protein, is prominently expressed during outflow tract (OFT) morphogenesis as a component of both the endocardial cushions and OFT valves. Here we sought to investigate the role of Fbln1 in the process of OFT valvulogenesis. Since Fbln1 null mice die perinatally, we first evaluated the effects of Fbln1 deficiency on OFT valve morphology in hearts of embryos at E17.5. Morphometric analysis of 3D reconstructions revealed that in Fbln1 nulls the volume of the anterior leaflet of the pulmonary valve, and the left and non-coronary leaflets of the aortic valve were significantly reduced as compared to wild type (WT) (30-40% reduction; WT n=6, null n=8; p<0.05). The reduced leaflet size correlated with decreased leaflet cell numbers (45%, 30% and 30% respectively; p<0.05). TUNEL and BrdU analysis showed no difference between E17.5 WT and Fbln1 null OFT valves. To determine when abnormalities of OFT valve leaflet cell numbers first occurs, OFT endocardial cushions and prevalvular mesenchyme from E10.5-14.5 null and WT embryos were analyzed. At E10.5, the proximal OFT cushions of Fbln1 nulls were hypercellular (34% increase; p<0.05; n=5 for each genotype) whereas cellularity of distal cushions was not different from WT (p= 0.49). The fact that the ratio of proximal OFT cushion mesenchymal cells to endothelial cells was increased in E10.5 Fbln1 null cushions suggests that alteration in EMT might contribute to the hypercellularity. At E12.5, cellularity of the proximal OFT cushions was observed to decrease by 35% (p<0.05; WT n=4; null n=4). This was accompanied by a 172% increase in apoptosis, with no alteration in the level of proliferation (p<0.05; WT n=3; null n=4). Decreased cellularity and increased apoptosis persisted in the OFT prevalvular mesenchyme of Fbln1 nulls at E14.5 (p<0.05; WT n=2; null n=2). Taken together, Fbln1 specifically regulates cellularization of the proximal endocardial cushion and prevalvular mesenchyme. Mechanistically, Fbln1 may act as a suppressor of cushion EMT and indirectly or directly as an inhibitor of apoptosis.

scholarly journals ANRIL Regulates Multiple Molecules of Pathogenetic Significance in Diabetic Nephropathy

2021 ◽  
Author(s):  
Parisa Sooshtari ◽  
Biao Feng ◽  
Saumik Biswas ◽  
Michael Levy ◽  
Hanxin Lin ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Structural Damage ◽  
Matrix Protein ◽  
Body Weight Gain ◽  
Extracellular Matrix Protein ◽  
Wild Type ◽  
Reduced Body ◽  
In The Wild ◽  
Pathway Analyses ◽  
Stage Renal Disease

Abstract Background: Hyperglycemia-induced transcriptional alterations lead to aberrant synthesis of a large number of pathogenetic molecules leading to functional and structural damage to multiple end organs including the kidneys. Diabetic nephropathy (DN) remains a major cause of end stage renal disease. Multiple epigenetic mechanisms, including alteration of long non-coding RNAs (lncRNAs) may play a significant role mediating the cellular transcriptional activities. We have previously shown that lncRNA ANRIL may mediate diabetes associated molecular, functional and structural abnormalities in DN. Here we explored downstream mechanisms of ANRIL alteration in DN.Methods: We used renal cortical tissues from ANRIL knockout (KO) mice and wild type (WT) B6 mice, with or without streptozotocin (STZ) induced diabetes for RNA sequencing. The differentially expressed genes were identified using edgeR and DESeq2 computational methods. KEGG and Reactome pathway analyses and network analysesusing STRING and IPA were subsequently performed. Results: Diabetic animals showed hyperglycemia, reduced body weight gain, polyuria and increased urinary albumin. Both albuminuria and polyuria were corrected in the KO diabetic mice. RNA analyses showed Diabetes induced alterations of a large number of transcripts in the wild type (WT) animals. ANRIL knockout (KO) prevented a large number of such alterations. The altered transcripts include metabolic pathways, apoptosis, extracellular matrix protein synthesis and degradation, NFKB related pathways, AGERAGE interaction pathways etc. ANRIL KO prevented majority of these pathways. Conclusion: These findings suggest that as ANRIL regulates a large number of molecules of pathogenetic significance, it may potentially be a drug target for DN and other chronic diabetic complications.

scholarly journals Kidney-specific WNK1 regulates sodium reabsorption and potassium secretion in mouse cortical collecting duct

2013 ◽  
Vol 304 (4) ◽  
pp. F397-F402 ◽  
Author(s):  
Chih-Jen Cheng ◽  
Michel Baum ◽  
Chou-Long Huang
Keyword(s):  
Collecting Duct ◽  
Sodium Reabsorption ◽  
Thick Ascending Limb ◽  
Cortical Collecting Duct ◽  
Wild Type ◽  
High K ◽  
In The Wild ◽  
Potassium Secretion ◽  
And Control

Kidney-specific with-no-lysine kinase 1 (KS-WNK1) is a kinase-deficient variant of WNK1 that is expressed exclusively in the kidney. It is abundantly expressed in the distal convoluted tubule (DCT) and to a lesser extent in the cortical thick ascending limb (cTAL), connecting tubule, and cortical collecting duct (CCD). KS-WNK1 inhibits Na+-K+-2Cl−- and sodium chloride cotransporter-mediated Na+ reabsorption in cTAL and DCT, respectively. Here, we investigated the role of KS-WNK1 in regulating Na+ and K+ transport in CCD using in vitro microperfusion of tubules isolated from KS-WNK1 knockout mice and control wild-type littermates. Because baseline K+ secretion and Na+ reabsorption were negligible in mouse CCD, we studied tubules isolated from mice fed a high-K+ diet for 2 wk. Compared with that in wild-type tubules, K+ secretion was reduced in KS-WNK1 knockout CCD perfused at a low luminal fluid rate of ∼1.5 nl/min. Na+ reabsorption and the lumen-negative transepithelial potential difference were also lower in the KS-WNK1 knockout CCD compared with control CCD. Increasing the perfusion rate to ∼5.5 nl/min stimulated K+ secretion in the wild-type as well as knockout CCD. The magnitudes of flow-stimulated increase in K+ secretion were similar in wild-type and knockout CCD. Maxi-K+ channel inhibitor iberiotoxin had no effect on K+ secretion when tubules were perfused at ∼1.5 nl/min, but completely abrogated the flow-dependent increase in K+ secretion at ∼5.5 nl/min. These findings support the notion that KS-WNK1 stimulates ROMK-mediated K+ secretion, but not flow-dependent K+ secretion mediated by maxi-K+ channels in CCD. In addition, KS-WNK1 plays a role in regulating Na+ transport in the CCD.

scholarly journals SEX-SPECIFIC TRAJECTORIES OF REELIN-DEPENDENT MATURATION OF DEEP LAYER PREFRONTAL NEURONS

2021 ◽  
Author(s):  
Thenzing J Silva Hurtado ◽  
Olivier Lassalle ◽  
Antoine Ameloot ◽  
Pascale Chavis
Keyword(s):  
Matrix Protein ◽  
Pyramidal Neurons ◽  
Deep Layer ◽  
Long Term Potentiation ◽  
Developmental Trajectory ◽  
Extracellular Matrix Protein ◽  
Wild Type ◽  
Functional Maturation

Throughout early adulthood, the anatomical and functional maturation of PFC circuitry continues under the influence of multiple extrinsic and intrinsic factors, most notably electrical activity, and molecular cues. We previously showed that the extracellular matrix protein reelin orchestrates the structural and functional maturation of deep layers medial PFC (mPFC) pyramidal neurons. Additionally, we reported that reelin haploinsufficiency is associated to prefrontal disruptions of long-term memory retention thereby illustrating the eminent role of reelin in cognitive maturation of the PFC. Prefrontal maturation follows a sex-specific developmental pattern, supporting the existence of sexual differences in the morphology-functional properties PFC. Here, we interrogated the role of reelin in the functional maturation of excitatory networks in the mPFC. The developmental trajectory of reelin's expression and deep layer pyramidal neurons synaptic plasticity was tracked in the mPFC of male and female mice, from the juvenile period to adulthood. To assess the role of reelin in both sexes, wild-type and heterozygous reeler mice (HRM) were compared. The results show that the maturational profile of reelin expression in the mPFC is sex-dependent and that the developmental trajectory of long-term potentiation is different between wild-type males and females. These data demonstrate reelin influence on prefrontal synapses is sex and period specific.

scholarly journals Kidney-specific WNK1 inhibits sodium reabsorption in the cortical thick ascending limb

2012 ◽  
Vol 303 (5) ◽  
pp. F667-F673 ◽  
Author(s):  
Chih-Jen Cheng ◽  
Thao Truong ◽  
Michel Baum ◽  
Chou-Long Huang
Keyword(s):  
Collecting Duct ◽  
Urine Volume ◽  
Sodium Reabsorption ◽  
Renal Tubules ◽  
Thick Ascending Limb ◽  
Cortical Collecting Duct ◽  
Wild Type ◽  
High K

Kidney-specific WNK1 (KS-WNK1) is a variant of full-length WNK1. Previous studies have reported that KS-WNK1 is predominantly expressed in the distal convoluted tubule (DCT) where it regulates sodium-chloride cotransporter. The role of KS-WNK1 in other nephron segments is less clear. Here, we measured the expression of KS-WNK1 transcript in microdissected renal tubules and found that KS-WNK1 was most abundant in the DCT, followed by cortical thick ascending limb (cTAL), connecting tubule, and cortical collecting duct. A high K+ diet enhanced the expression of KS-WNK1 in the DCT and cTAL, selectively. It has been reported that a high-K diet suppresses Na+ reabsorption in TAL. To understand the role of KS-WNK1 in Na+ transport in cTAL and the regulation by dietary K+, we examined Na+ reabsorption using in vitro microperfusion in cTAL isolated from KS-WNK1-knockout mice and wild-type littermates fed either a control-K+ or high-K+ diet. Furosemide-sensitive Na+ reabsorption in cTAL was higher in KS-WNK1-knockout (KO) mice than in wild-type. A high-K+ diet inhibited Na+ reabsorption in cTAL from wild-type mice, but the inhibition was eliminated in KS-WNK1-KO mice. We further examined the role of KS-WNK1 using transgenic mice that overexpress KS-WNK1. Na+ reabsorption in cTAL was lower in transgenic than in wild-type mice. In whole animal clearance studies, a high-K+ diet increased daily urine volume and urinary Na+ and K+ excretion in wild-type mice, which was blunted in KS-WNK1-KO mice. Thus KS-WNK1 inhibits Na+ reabsorption in cTAL and mediates the inhibition of Na+ reabsorption in the segment by a high-K diet.

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