Prolonged hyperglycemia leads to diabetes complications, especially in individuals with African ancestry (AFR) - a health disparity. Glucose-6-phosphate dehydrogenase deficiency (G6PDd) ...disproportionately affects men with AFR ancestry (prevalence 9.5% vs. 2.2% in the US population) and shortens red cell lifespan, reducing HbA1c with no effect on glucose levels. We investigated whether men with AFR ancestry and G6PDd were at increased risk of diabetes complications. We performed a multi-ethnic genome-wide association study meta-analysis of diabetic retinopathy (DR) using the VA Million Veteran Program (MVP) dataset (nmax = 192,406) and studied clinical impact with the MVP and ACCORD trial datasets. Nine significant loci were associated with DR, including a causal variant for G6PDd rs1050828-T, OR 1.48 (95% CI 1.45 - 1.51), p = 1.99x10-90). Plasma glucose was much higher in those with vs. without G6PDd in the year preceding diabetes diagnosis (168 vs. 137 mg/dL, respectively) and insulin prescription (253 vs. 233), both p <0.001. In a Cox proportional hazards analysis, ACCORD participants with vs. without G6PDd had a higher likelihood of DR HR 1.78 (1.55 - 2.04), p <0.001 and neuropathy (HR 1.37 (1.23 - 1.54), p <0.005. A mediation analysis of G6PDd on DR showed that risk was fully attributable to higher glucose levels. In MVP participants, compared to those without G6PDd who were in the top two tertiles of HbA1c regressed onto plasma glucose, the risk of DR was increased with G6PDd (OR 1.40), but lower than the risk of DR in those without G6PDd who were in the lowest tertile of A1c vs. glucose (OR 1.61) - consistent with risk due to inadequate treatment rather than oxidative stress alone. Conclusions: Management based on both glucose and HbA1c, rather than HbA1c alone, might be needed to reduce diabetes complications in both individuals with African ancestry who have G6PDd, and other individuals with low HbA1c levels relative to their glucose levels. Disclosure J.H. Breeyear: None. J. Hellwege: None. J.S. House: None. S.L. Mitchell: None. B. Charest: None. T.B. Basnet: None. P. Reaven: Research Support; Dexcom, Inc. J.B. Meigs: None. M.K. Rhee: Research Support; Kowa Pharmaceuticals America, Inc. Y. Sun: None. O. Wilson: None. A.M. Hung: None. S.K. Iyengar: None. D.M. Rotroff: Consultant; Novo Nordisk. Research Support; Bayer Inc. J.B. Buse: Other Relationship; Novo Nordisk. Consultant; Corcept Therapeutics. Research Support; Corcept Therapeutics, Dexcom, Inc., Insulet Corporation. Consultant; Alkahest, Anji Pharmaceuticals, Aqua Medical, Altimmune Inc., AstraZeneca, Boehringer-Ingelheim, CeQur, Eli Lilly and Company, embecta, GentiBio, Glyscend Inc., Mellitus Health, Metsera, Pendulum Therapeutics, Praetego, LLC, Stability Health, Terns Pharmaceuticals, Insulet Corporation, Vertex Pharmaceuticals Incorporated, vTv Therapeutics. Other Relationship; Medtronic. Stock/Shareholder; Glyscend Inc., Mellitus Health, Pendulum Therapeutics, Praetego, LLC, Stability Health. A. Leong: Other Relationship; Merck & Co., Inc. J.M. Mercader: None. M. Brantley: None. N.S. Peachey: None. A. Motsinger-Reif: None. P.W. Wilson: None. Y. Sun: None. A. Giri: None. L.S. Phillips: Other Relationship; Diasyst, Inc. Research Support; Kowa Pharmaceuticals America, Inc., Janssen Pharmaceuticals, Inc., AbbVie Inc., Novo Nordisk, GlaxoSmithKline plc, Abbott, Sanofi-Aventis U.S., Pfizer Inc. T.L. Edwards: None. Funding NEI (F31EY033663, T32EY021453-10, R01EY025295, R01EY032159, P30-EY026877), NICHD (K12HD043483), NIAMS (K12AR084232-24), NIDDK (R01DK127083, K01DK120631, R21AI156161), NHGRI (U01HG011723, UL1TR002378)
IntroductionHydrochlorothiazide (HCTZ) is among the most commonly prescribed antihypertensives in the US, yet <50% of HCTZ treated patients achieve blood pressure (BP) control.HypothesisIntegrating ...metabolomic and genomic profiles of HCTZ treated patients could identify novel pathways/biomarkers for optimizing the use of HCTZ.MethodsPrimary analysis included 228 white hypertensives treated with HCTZ from the Pharmacogenomic Evaluation of Antihypertensive Response (PEAR) study. Genotyping was determined via Illumina Omni 1M-Quad Chip and untargeted metabolomics was performed on baseline fasting plasma samples using a GC-TOF MS platform. Pathway analysis was used to integrate metabolites significantly associated with HCTZ BP response (FDR<.05) and genetic signals at p<5x10-5 from HCTZ BP genome-wide analysis. Replication was conducted in another 212 white HCTZ treated patients. Replicated genetic signals were used to create a BP response score by summing their BP lowering alleles. We validated this score by testing it in 196 white HCTZ treated patients in the Genetic Epidemiology of Responses to Antihypertensives (GERA) study.ResultsThirteen metabolites were significantly associated with HCTZ systolic BP (SBP) and diastolic BP (DBP) responses (FDR<.05). Metabolomics-genomics integration revealed significant metabolites along with rs2727563 PRKAG2, rs12604940 DCC, and rs13262930 EPHX2, in the netrin signalling pathway (p=1x10-5), as potential markers significantly influencing HCTZ BP response. We successfully replicated those 3 genetic signals and added their alleles to create a response score which explained 11.3% and 11.9% of the variability to HCTZ SBP and DBP responses, respectively. We found that patients carrying one “response” allele had a significantly worse response than carriers of six alleles (ΔSBP/ΔDBP-1.5/1.2 vs -16.3/-10.4 mmHg, respectively, SBP score p=1x10-8 and DBP score p=3x10-9). We further validated this response score by testing it in the GERA (DBP score p=0.03, SBP score p=0.07).ConclusionThis study highlights the power of using different omics to identify novel pathways/biomarkers of drug response, and suggests that PRKAG2, DCC and EPHX2 might be important determinants of HCTZ BP response.
The AGMK1-9T7 cell line has been used to study neoplasia in tissue culture. By passage in cell culture, these cells evolved to become tumorigenic and metastatic in immunodeficient mice at passage 40. ...Of the 20 x 106 kidney cells originally plated, less than 2% formed the colonies that evolved to create this cell line. These cells could be the progeny of some type of kidney progenitor cells. To characterize these cells, we documented their renal lineage by their expression of PAX-2 and MIOX, detected by indirect immunofluorescence. These cells assessed by flow-cytometry expressed high levels of CD44, CD73, CD105, Sca-1, and GLI1 across all passages tested; these markers have been reported to be expressed by renal progenitor cells. The expression of GLI1 was confirmed by immunofluorescence and western blot analysis. Cells from passages 13 to 23 possessed the ability to differentiate into adipocytes, osteoblasts, and chondrocytes; after passage 23, their ability to form these cell types was lost. These data indicate that the cells that formed the AGMK1-9T7 cell line were GLI1+ perivascular, kidney, progenitor cells.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
To study neoplasia in tissue culture, cell lines representing the evolution of normal cells to tumor cells are needed. To produce such cells, we developed the AGMK1-9T7 cell line, established cell ...banks at 10-passage intervals, and characterized their biological properties. Here we examine the evolution of chromosomal DNA copy-number aberrations and miRNA expression in this cell line from passage 1 to the acquisition of a tumorigenic phenotype at passage 40. We demonstrated the use of a human microarray platform for DNA copy-number profiling of AGMK1-9T7 cells using knowledge of synteny to ‘recode’ data from human chromosome coordinates to those of the African green monkey. This approach revealed the accumulation of DNA copy-number gains and losses in AGMK1-9T7 cells from passage 3 to passage 40, which spans the period in which neoplastic transformation occurred. These alterations occurred in the sequences of genes regulating DNA copy-number imbalance of several genes that regulate endothelial cell angiogenesis, survival, migration, and proliferation. Regarding miRNA expression, 195 miRNAs were up- or down-regulated at passage 1 at levels that appear to be biologically relevant (i.e., log2 fold change >2.0 (q<0.05)). At passage 10, the number of up/down-regulated miRNAs fell to 63; this number increased to 93 at passage 40. Principal-component analysis grouped these miRNAs into 3 clusters; miRNAs in sub-clusters of these groups could be correlated with initiation, promotion, and progression, stages that have been described for neoplastic development. Thirty-four of the AGMK1-9T7 miRNAs have been associated with these stages in human cancer. Based on these data, we propose that the evolution of AGMK1-9T7 cells represents a detailed model of neoplasia in vitro.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
PDF
