19 research outputs found
D50A ve A88V mutasyonlarına bağlı connexin26 protein lokalizasyonunun ve hücre içi kalsiyum miktarının değişimi
No full textIntroduction: Connexins (Cx) play essential roles in cellular homeostasis by forming gap junctions and non-junctional hemichannels. In vitro characterization of Cx26 mutations causing keratitis-ichthyosis-deafness (KID) syndrome, were shown to form leaky hemichannels. The molecular/ cellular mechanisms affected by aberrant hemichannels have recently been elucidated. Here, we further wanted to characterize Cx26 KID syndrome mutations, D50A and A88V, which were shown to form aberrant hemichannels and remained unaddressed in the literature. Methods: Neurobiotin uptake assay in HeLa and N2A cells transfected with Cx26-WT, D50A or A88V verified the presence of aberrant hemichannels and immunofluorescent staining with fluorescent microscopy determined cellular localization of Cx26. Finally, intracellular calcium content was examined by using calcium indicator, Fluo-3AM, and flow cytometer. Results: Cx26-D50A and A88V mutations prevented the formation of gap junction plaques at cell-cell appositions and mutant proteins were observed to localize to the Golgi apparatus. Further, comparison of intracellular calcium content showed an increase in calcium amount in cells containing Cx26-D50A and A88V relative to Cx26-WT. Conclusion: Retention of Cx26 in the Golgi apparatus and alteration in the intracellular calcium content due to KID syndrome mutations may influence various cellular processes that might contribute to development of epidermal phenotypes.Connexin molekülleri (Cx) kurdukları hücrelerarası
kanallar ve hücrelerarası olmayan yarı kanallar sayesinde
hücre yaşamını dengelemede önemli roller üstlenirler. İn
vitro analizlerle keratitis-ichthyosis-deafness (KID) sendromuyla
ilişkili Cx26 mutasyonlarının düzgün kapanamayan
yarı kanalların oluşmasına sebep oldukları gösterilmiştir.
Bu tip anormal çalışan yarı kanalların etkilediği
moleküler / hücresel mekanizmalar son zamanlarda araştırılmaya
başlanmıştır. Bu çalışmada amacımız, daha
önce yarı kanallar yaptıkları gösterilen Cx26’da oluşan
KID sendromu mutasyonlarından D50A ve A88V’nin
hücre biyolojisinde yaptığı değişiklikleri incelemektir.
Yöntemler: Cx26-WT, D50Y ve A88V ile transfekte edilen
HeLa ve N2A hücrelerinde neurobiotin boya alım deneyleriyle
anormal yarı kanalların varlığı teyit edilmiş ve floresan
mikroskop ve immunoboyamasıyla Cx26’nın hücre
içi lokalizayonu belirlenmiştir. Son olarak, kalsiyum belirteci,
Fluo-3AM ve akış sitometresi kullanarak hücre içi kalsiyum
içeriği belirlenmiştir.Scientific and Technological Research Council of Turkey (210T035); FP7 Marie Curie Re-Integration (PIRG08-GA-2010-277101
Altered cellular localization and hemichannel activities of KID syndrome associated connexin26 I30N and D50Y mutations
No full textBackground: Gap junctions facilitate exchange of small molecules between adjacent cells, serving a crucial function for the maintenance of cellular homeostasis. Mutations in connexins, the basic unit of gap junctions, are associated with several human hereditary disorders. For example, mutations in connexin26 (Cx26) cause both non-syndromic deafness and syndromic deafness associated with skin abnormalities such as keratitis-ichthyosis-deafness (KID) syndrome. These mutations can alter the formation and function of gap junction channels through different mechanisms, and in turn interfere with various cellular processes leading to distinct disorders. The KID associated Cx26 mutations were mostly shown to result in elevated hemichannel activities. However, the effects of these aberrant hemichannels on cellular processes are recently being deciphered. Here, we assessed the effect of two Cx26 mutations associated with KID syndrome, Cx26I30N and D50Y, on protein biosynthesis and channel function in N2A and HeLa cells.Results: Immunostaining experiments showed that Cx26I30N and D50Y failed to form gap junction plaques at cell-cell contact sites. Further, these mutations resulted in the retention of Cx26 protein in the Golgi apparatus. Examination of hemichannel function by fluorescent dye uptake assays revealed that cells with Cx26I30N and D50Y mutations had increased dye uptake compared to Cx26WT (wild-type) containing cells, indicating abnormal hemichannel activities. Cells with mutant proteins had elevated intracellular calcium levels compared to Cx26WT transfected cells, which were abolished by a hemichannel blocker, carbenoxolone (CBX), as measured by Fluo-3 AM loading and flow cytometry.Conclusions: Here, we demonstrated that Cx26I30N and D50Y mutations resulted in the formation of aberrant hemichannels that might result in elevated intracellular calcium levels, a process which may contribute to the hyperproliferative epidermal phenotypes of KID syndrome
Loss of P3h2 gene causes thin basement membrane nephropathy in mice
No full textGFB is the decisive filter to generate primary urine. It is composed of a fenestrated endothelium, GBM, podocytes and SD. Secreted proteins (Col4, LM-521, nidogen, and agrin) from GECs and podocytes generate the meshwork structure of the GBM. It works as a scaffold to support these two cells and provides cellular cross-talk to keep the dynamic structure of the GFB in homeostasis. Disruption of the GFB leads to proteinuria and microhematuria.
A P3H2 gene mutation was identified in a nephrotic syndrome patient cohort by using an expression-based candidate gene approach. P3H2 encodes Prolyl 3-Hydroxylase 2 (P3H2), a post-translational Col4 modifier that hydroxylases the 3’ of prolines in Col4 sub-chains in the ER. P3H2’s relevance for glomerular homeostasis is unknown. Therefore, we aimed to investigate its effect on GBM and GFB homeostasis. We generated P3h2∆Pod mice line and P3H2 KO immortalized human podocyte cell lines. Characterization of these mice revealed that the absence of P3H2 in podocytes induces TBMN phenotype with thinner GBM than wild type mice and the development of microhematuria and microalbuminuria over time. Sclerosis in glomeruli, podocyte loss and podocyte hypertrophy were observed at 48w of age in KO mice. There was FSGS development in P3h2∆Pod mice over time. ECM and GBM proteomics of P3H2 KO cell lines and mice indicated a disrupted in the regulation of Col4 sub-chains and its interaction partners (Col18a1 and nidogens).
In conclusion, P3H2 is the first identified GBM modulator causing TBMN phenotype in mice and inducing FSGS over time. P3H2 might affect proper Col4 biosynthesis or stability explaining the observed phenotype on a molecular level.Die GFB ist der entscheidende Filter zur Erzeugung von Primärurin. Sie besteht aus einem fenestrierten Endothel, GBM, Podozyten und der SD. Sezernierte Proteine (Col4, LM-521, Nidogen und Agrin) aus GECs und Podozyten erzeugen die Netzstruktur der GBM. Sie fungiert als Gerüst zur Unterstützung dieser beiden Zelltypen und vermittelt zellulären Crosstalk, um die dynamische Struktur der GFB in der Homöostase zu halten. Eine Störung der GFB führt zu Proteinurie und Mikrohämaturie.
Eine P3H2-Genmutation wurde in einer Patientenkohorte mit nephrotischem Syndrom unter Verwendung eines expressions-basierten Kandidatengenansatzes identifiziert. P3H2 codiert Prolyl-3-Hydroxylase 2 (P3H2), einen posttranslationalen Col4 modifikator, der die 3'-Proline in Col4-Subketten im endoplasmatischen Retikulum hydroxyliert. Die Relevanz von P3H2 für die glomeruläre Homöostase ist unbekannt. Daher wollten wir die Auswirkungen auf die GBM- und GFB-Homöostase untersuchen. Wir erzeugten eine P3h2∆Pod-Mauslinie und P3H2-KO-immortalisierte menschliche Podozytenzelllinien. Die Charakterisierung dieser Mäuse ergab, dass das Fehlen von P3H2 in Podozyten einen progressiven Phänotyp der dünnen Basalmembran-Nephropathie (TBMN) mit dünnerem GBM als bei Wildtyp-Mäusen und die Entwicklung von Mikrohämaturie und Mikroalbuminurie im Laufe der Zeit induziert. Sklerose von Glomeruli, Podozytenverlust und Podozytenhypertrophie wurden im Alter von 48 W von KO-Mäusen beobachtet. Im Laufe der Zeit gab es eine FSGS-Entwicklung bei P3h2∆Pod-Mäusen. Die ECM- und GBM-Proteomik von P3H2-KO-Zelllinien und -Mäusen zeigte eine gestörte Regulation der Col4-Subketten und ihrer Interaktionspartner (Col18a1 und nidogene).
Zusammenfassend ist P3H2 der erste identifizierte GBM-Modulator, der bei Mäusen einen TBMN-Phänotyp verursacht und im Laufe der Zeit FSGS induziert. P3H2 könnte die ordnungsgemäße Col4-Biosynthese oder -Stabilität beeinflussen und den beobachteten Phänotyp auf molekularer Ebene erklären
Connexin 26- I30N ve D50Y mutasyonlarının sebep olduğu normal çalışmayan yarım kanalların fonksiyonlarının belirlenmesi
No full textThesis (Master)--Izmir Institute of Technology, Molecular Biology and Genetics, Izmir, 2015Includes bibliographical references (leaves: 42-49)Text in English; Abstract: Turkish and Englishix, 49 leavesCells need to communicate with each other for maintenance cellular and tissue homeostasis. Gap junctions are channel-forming structures that are formed by docking of two hemichannels on the plasma membrane of adjacent cells. Connexins are subunits of gap junctions. Connexin 26 (Cx26) is one of the connexin isoform and mutations on the Cx26 gene (GJB2) cause non-syndromic and syndromic deafness. Keratitis-ichthyosis-deafness (KID) syndrome is one of the syndromic deafness disorders caused by Cx26 mutations. Among these mutations, Cx26-I30N and D50Y missense mutations were shown to form aberrant hemichannels but their effect on protein biosynthesis and functions have not studied. In this study, we aimed to decipher in vitro functions of aberrant hemichannels formed by Cx26-I30N and D50Y mutations.
First of all, the effect of Cx26-I30N and D50Y mutations on localization, mRNA expression and protein synthesis properties were investigated in HeLa, N2A and HaCaT cells. Results suggested that Cx26-I30N and D50Y mutants were not able to form gap junction plaques on the plasma membrane and were localized in the Golgi apparatus. In addition, mutations resulted in a reduction in mRNA expression and protein synthesis. After, functional analysis was performed in Cx26-I30N and D50Y transfected N2A and HaCaT cells. Internal Ca2+ content measurement, measurement of released ATP, measurement of cell size and apoptosis assays were performed. Ca2+ measurement results showed that both Cx26-I30N and D50Y mutations deregulate Ca2+ balance in both N2A and HaCaT cells. Result of ATP release assay indicated that ATP amount in the extracellular environment decreased in N2A cells having Cx26-I30N and D50Y clones. Finally, apoptosis assay showed that number of necrotic cells increased when N2A cells were transfected with Cx26-I30N and D50Y constructs. Therefore, it was shown that aberrant hemichannels formed by Cx26-I30N and D50Y mutations may induce necrotic cell death by disrupting Ca2+ balance and ATP amount in cells.Hücresel ve doku dengeleşiminin devamı için hücreler arası iletişim şarttır. Oluklu bağlantılar kanal oluşturan yapılardır ve komşu hücrelerin, hücre zarındaki yarım kanalların birbirine bağlanması sonucu oluşurlar. Connexinler oluklu bağlnatıların yapı proteinleridir. Connexin 26 (Cx26) bir connexin formudur ve Cx26 geninde (GJB2) meydana gelen mutasyonlar sendromik ve sendromik olmayan sağirlığa sebep olur. Keratitis-ichthyosis-deafness (KID) Syndrome Cx26 gene mutasyonlarından dolayı meydana gelen bir sendromic hastalıktır. Bu mutasyonlardan Cx26-I30N ve D50Y yanlış anlam mutasyonları anormal çalışan yarım kanal oluşmasına sebep olmuşlardır. Fakat anormal çalışan kanalların protein biyosentezine ve fonksiyonuna etkileri bilinmemektedir. Bu çalışmada, Cx26-I30N ve D50Y mutasyonları tarafından oluşturulan yarım kanalların in vitro ortamda fonksiyonlarının belirlenmesi hedeflenmiştir.
Ilk olarak, Cx26-I30N ve D50Y mutasyonlarının hücre içi lokalizasyon, mRNA ifadelenmesi ve protein sentez miktarlarına etkileri HeLa, N2A ve HaCaT hücrelerinde belirlenmiştir. Sonuçlarda, Cx26-I30N ve D50Y mutantları oluklu bağlantı oluşturamazken, Golgi aygıtında birikmiştir. Ayrıca, mutantların mRNA ifadeleri ve protein sentez miktarları düşmüştür. Daha sonra, Cx26-I30N ve D50Y mutasyonlarının fonksiyonel analizleri N2A ve HaCaT hücrelerinde yapılmıştır. Hücre içi Ca2+ ölçümü, salınmış ATP ölçümü, hücre boyutu ölçümü ve apoptosiz tahlili yapılan deneylerdendir. Ca2+ ölçümü göstermiştir ki Cx26-I30N and D50Y mutasyonları Ca2+ dengesini hem N2A hem de HaCaT hücrelerinde bozmaktadir. Ek olarak, salınmış ATP deneyi göstermiştir ki, hücre çevresindeki salınmış ATP miktarı içinde Cx26-I30N ve D50Y klonları olan N2A hücrelerinde düşmüştür. Son olarak yapılan apoptoziz deneyine göre, Cx26-I30N ve D50Y mutasyonlarını içeren N2A hücreleri nekroz olmuşlardır. Sonuçlar göstermiştir ki, anormal çalışan yarim kanallar Ca2+ ve ATP dengelerini bozarak, hücrelerin nekroz olmasına sebep olmuştur.TÜBİTAK: Grant number 210T03
Connexin 26- I30N ve D50Y mutasyonlarının sebep olduğu normal çalışmayan yarım kanalların fonksiyonlarının belirlenmesi
No full textThesis (Master)--Izmir Institute of Technology, Molecular Biology and Genetics, Izmir, 2015Includes bibliographical references (leaves: 42-49)Text in English; Abstract: Turkish and Englishix, 49 leavesCells need to communicate with each other for maintenance cellular and tissue homeostasis. Gap junctions are channel-forming structures that are formed by docking of two hemichannels on the plasma membrane of adjacent cells. Connexins are subunits of gap junctions. Connexin 26 (Cx26) is one of the connexin isoform and mutations on the Cx26 gene (GJB2) cause non-syndromic and syndromic deafness. Keratitis-ichthyosis-deafness (KID) syndrome is one of the syndromic deafness disorders caused by Cx26 mutations. Among these mutations, Cx26-I30N and D50Y missense mutations were shown to form aberrant hemichannels but their effect on protein biosynthesis and functions have not studied. In this study, we aimed to decipher in vitro functions of aberrant hemichannels formed by Cx26-I30N and D50Y mutations.
First of all, the effect of Cx26-I30N and D50Y mutations on localization, mRNA expression and protein synthesis properties were investigated in HeLa, N2A and HaCaT cells. Results suggested that Cx26-I30N and D50Y mutants were not able to form gap junction plaques on the plasma membrane and were localized in the Golgi apparatus. In addition, mutations resulted in a reduction in mRNA expression and protein synthesis. After, functional analysis was performed in Cx26-I30N and D50Y transfected N2A and HaCaT cells. Internal Ca2+ content measurement, measurement of released ATP, measurement of cell size and apoptosis assays were performed. Ca2+ measurement results showed that both Cx26-I30N and D50Y mutations deregulate Ca2+ balance in both N2A and HaCaT cells. Result of ATP release assay indicated that ATP amount in the extracellular environment decreased in N2A cells having Cx26-I30N and D50Y clones. Finally, apoptosis assay showed that number of necrotic cells increased when N2A cells were transfected with Cx26-I30N and D50Y constructs. Therefore, it was shown that aberrant hemichannels formed by Cx26-I30N and D50Y mutations may induce necrotic cell death by disrupting Ca2+ balance and ATP amount in cells.Hücresel ve doku dengeleşiminin devamı için hücreler arası iletişim şarttır. Oluklu bağlantılar kanal oluşturan yapılardır ve komşu hücrelerin, hücre zarındaki yarım kanalların birbirine bağlanması sonucu oluşurlar. Connexinler oluklu bağlnatıların yapı proteinleridir. Connexin 26 (Cx26) bir connexin formudur ve Cx26 geninde (GJB2) meydana gelen mutasyonlar sendromik ve sendromik olmayan sağirlığa sebep olur. Keratitis-ichthyosis-deafness (KID) Syndrome Cx26 gene mutasyonlarından dolayı meydana gelen bir sendromic hastalıktır. Bu mutasyonlardan Cx26-I30N ve D50Y yanlış anlam mutasyonları anormal çalışan yarım kanal oluşmasına sebep olmuşlardır. Fakat anormal çalışan kanalların protein biyosentezine ve fonksiyonuna etkileri bilinmemektedir. Bu çalışmada, Cx26-I30N ve D50Y mutasyonları tarafından oluşturulan yarım kanalların in vitro ortamda fonksiyonlarının belirlenmesi hedeflenmiştir.
Ilk olarak, Cx26-I30N ve D50Y mutasyonlarının hücre içi lokalizasyon, mRNA ifadelenmesi ve protein sentez miktarlarına etkileri HeLa, N2A ve HaCaT hücrelerinde belirlenmiştir. Sonuçlarda, Cx26-I30N ve D50Y mutantları oluklu bağlantı oluşturamazken, Golgi aygıtında birikmiştir. Ayrıca, mutantların mRNA ifadeleri ve protein sentez miktarları düşmüştür. Daha sonra, Cx26-I30N ve D50Y mutasyonlarının fonksiyonel analizleri N2A ve HaCaT hücrelerinde yapılmıştır. Hücre içi Ca2+ ölçümü, salınmış ATP ölçümü, hücre boyutu ölçümü ve apoptosiz tahlili yapılan deneylerdendir. Ca2+ ölçümü göstermiştir ki Cx26-I30N and D50Y mutasyonları Ca2+ dengesini hem N2A hem de HaCaT hücrelerinde bozmaktadir. Ek olarak, salınmış ATP deneyi göstermiştir ki, hücre çevresindeki salınmış ATP miktarı içinde Cx26-I30N ve D50Y klonları olan N2A hücrelerinde düşmüştür. Son olarak yapılan apoptoziz deneyine göre, Cx26-I30N ve D50Y mutasyonlarını içeren N2A hücreleri nekroz olmuşlardır. Sonuçlar göstermiştir ki, anormal çalışan yarim kanallar Ca2+ ve ATP dengelerini bozarak, hücrelerin nekroz olmasına sebep olmuştur.TÜBİTAK: Grant number 210T03
Keratitis-Ichthyosis Syndrome Associated Mutations Impair the Localization and Functions of Connexin 26
No full textConnexins (Cx) form gap junctions and non-junctional hemichannels that play roles in several cellular mechanisms, including proliferation and differentiation. The importance of connexins for human physiology was shown by the association of mutations in several isoforms with various human hereditary disorders. Mutations in Cx26 cause both non-syndromic and syndromic deafness associated with skin disorders including keratitis-ichthyosis-deafness (KID) syndrome. In vitro characterization of Cx26 mutations suggested that mutations causing non-syndromic deafness and syndromic deafness show different properties, where the former ones result in loss-of-function and the latter ones cause gain-of-function mutations
Alteration of Protein Localization and Intracellular Calcium Content Due To Connexin26 D50a and A88v Mutations
No full textConnexin molekülleri (Cx) kurdukları hücrelerarası
kanallar ve hücrelerarası olmayan yarı kanallar sayesinde
hücre yaşamını dengelemede önemli roller üstlenirler. İn
vitro analizlerle keratitis-ichthyosis-deafness (KID) sendromuyla
ilişkili Cx26 mutasyonlarının düzgün kapanamayan
yarı kanalların oluşmasına sebep oldukları gösterilmiştir.
Bu tip anormal çalışan yarı kanalların etkilediği
moleküler / hücresel mekanizmalar son zamanlarda araştırılmaya
başlanmıştır. Bu çalışmada amacımız, daha
önce yarı kanallar yaptıkları gösterilen Cx26’da oluşan
KID sendromu mutasyonlarından D50A ve A88V’nin
hücre biyolojisinde yaptığı değişiklikleri incelemektir.
Yöntemler: Cx26-WT, D50Y ve A88V ile transfekte edilen
HeLa ve N2A hücrelerinde neurobiotin boya alım deneyleriyle
anormal yarı kanalların varlığı teyit edilmiş ve floresan
mikroskop ve immunoboyamasıyla Cx26’nın hücre
içi lokalizayonu belirlenmiştir. Son olarak, kalsiyum belirteci,
Fluo-3AM ve akış sitometresi kullanarak hücre içi kalsiyum
içeriği belirlenmiştir.Introduction: Connexins (Cx) play essential roles in cellular homeostasis by forming gap junctions and non-junctional hemichannels. In vitro characterization of Cx26 mutations causing keratitis-ichthyosis-deafness (KID) syndrome, were shown to form leaky hemichannels. The molecular/ cellular mechanisms affected by aberrant hemichannels have recently been elucidated. Here, we further wanted to characterize Cx26 KID syndrome mutations, D50A and A88V, which were shown to form aberrant hemichannels and remained unaddressed in the literature. Methods: Neurobiotin uptake assay in HeLa and N2A cells transfected with Cx26-WT, D50A or A88V verified the presence of aberrant hemichannels and immunofluorescent staining with fluorescent microscopy determined cellular localization of Cx26. Finally, intracellular calcium content was examined by using calcium indicator, Fluo-3AM, and flow cytometer. Results: Cx26-D50A and A88V mutations prevented the formation of gap junction plaques at cell-cell appositions and mutant proteins were observed to localize to the Golgi apparatus. Further, comparison of intracellular calcium content showed an increase in calcium amount in cells containing Cx26-D50A and A88V relative to Cx26-WT. Conclusion: Retention of Cx26 in the Golgi apparatus and alteration in the intracellular calcium content due to KID syndrome mutations may influence various cellular processes that might contribute to development of epidermal phenotypes
Alteration of protein localization and intracellular calcium content due to connexin26 D50A and A88V mutations
No full textAbstractIntroduction:Connexins (Cx) play essential roles in cellular homeostasis by forming gap junctions and non-junctional hemichannels. In vitro characterization of Cx26 mutations causing keratitis-ichthyosis-deafness (KID) syndrome, were shown to form leaky hemichannels. The molecular/cellular mechanisms affected by aberrant hemichannels have recently been elucidated. Here, we further wanted to characterize Cx26 KID syndrome mutations, D50A and A88V, which were shown to form aberrant hemichannels and remained unaddressed in the literature.Methods:Neurobiotin uptake assay in HeLa and N2A cells transfected with Cx26-WT, D50A or A88V verified the presence of aberrant hemichannels and immunofluorescent staining with fluorescent microscopy determined cellular localization of Cx26. Finally, intracellular calcium content was examined by using calcium indicator, Fluo-3AM, and flow cytometer.Results:Cx26-D50A and A88V mutations prevented the formation of gap junction plaques at cell-cell appositions and mutant proteins were observed to localize to the Golgi apparatus. Further, comparison of intracellular calcium content showed an increase in calcium amount in cells containing Cx26-D50A and A88V relative to Cx26-WT.Conclusion:Retention of Cx26 in the Golgi apparatus and alteration in the intracellular calcium content due to KID syndrome mutations may influence various cellular processes that might contribute to development of epidermal phenotypes.</jats:sec
Comparative validation of the Mayo Clinic imaging classification and PROPKD score for predicting ADPKD progression across diverse ethnic cohorts.
No full textBackground Autosomal dominant polycystic kidney disease (ADPKD) is a common cause of end-stage kidney disease (ESKD). In this retrospective multicenter cohort study, we aimed to evaluate the concordance, sensitivity and specificity of the predictive abilities of the Mayo Clinic Imaging Class (MCIC) and Predicting Renal Outcome in Polycystic Kidney Disease (PROPKD) score for progression to ESKD. Methods Data for adult ADPKD patients of diverse ethnicities were obtained from 4 renal centers (Saint Louis, Kayseri, Bursa, Istanbul). The predictive value of MCIC and PROPKD was compared for progression to ESKD. ESKD was defined by the following criteria: (1) two consecutive estimated glomerular filtration rate (eGFR) < 15 ml/min/1.73m(2) more than 1 month apart and/or (2) initiation of kidney replacement therapy. Results One hundred eighty-seven ADPKD patients with PKD1 or PKD2 variants were included, with the majority identifying as White (93%), followed by Black (6%) and Hispanic (0.5%). The PROPKD score showed moderate ability (AUC = 0.702) to predict progression to ESKD, performing better in Black patients (AUC = 0.800) than in White patients (AUC = 0.764). However, the predictive accuracy declined (AUC = 0.644) when the PROPKD score was categorized into risk levels (low, intermediate, high). The MCIC showed lower ability (AUC = 0.578) to predict progression to ESKD than the PROPKD score. Using LASSO regularization and multivariable Cox regression, self-identified Black ethnicity [HR = 12.24 (95% CI: 3.19-46.94, p < 0.001)] and age at initial follow-up [HR = 1.05 (95% CI:1.01-1.09, p = 0.007)] emerged as significant predictors of progression to ESKD. Conclusions The PROPKD score appears to be a useful tool for predicting ADPKD progression across various ethnic groups, with self-identified Black ethnicity and age at initial follow-up emerging as key predictors of kidney failure.[GRAPHICS].Saint Louis University Research Scholars in Internal Medicine Grant (RSIMG 2023
Genomics in Pancreas–Kidney Transplantation: From Risk Stratification to Personalized Medicine
No full textBackground: Pancreas and pancreas–kidney transplantation are well-established therapeutic options for patients with type 1 diabetes mellitus (T1DM) and end-stage kidney disease (ESKD), offering the potential to restore endogenous insulin production and kidney function. It improves metabolic control, quality of life, and long-term survival. While surgical techniques and immunosuppressive strategies have advanced considerably, graft rejection and limited long-term graft survival remain significant clinical challenges. Method: To better understand these risks, the genetic and immunological factors that influence transplant outcomes are examined. Beyond traditional human leukocyte antigen (HLA) matching, non-HLA genetic variants such as gene deletions and single-nucleotide polymorphisms (SNPs) have emerged as contributors to alloimmune activation and graft failure. Result: Polymorphisms in cytokine genes, minor histocompatibility antigens, and immune-regulatory pathways have been implicated in transplant outcomes. However, the integration of such genomic data into clinical practice remains limited due to underexplored gene targets, variability in study results, and the lack of large, diverse, and well-characterized patient cohorts. Initiatives like the International Genetics & Translational Research in Transplantation Network (iGeneTRAiN) are addressing these limitations by aggregating genome-wide data from thousands of transplant donors and recipients across multiple centers. These large-scale collaborative efforts aim to identify clinically actionable genetic markers and support the development of personalized immunosuppressive strategies. Conclusions: Overall, genetic testing and genomics hold great promise in advancing precision medicine in pancreas and pancreas–kidney transplantation
