An ACACB Variant Implicated in Diabetic Nephropathy Associates with Body Mass Index and Gene Expression in Obese Subjects

Ma, Lijun; Murea, Mariana; Snipes, James A.; Marinelarena, Alejandra; Krüger, Jacqueline; Hicks, Pamela J.; Langberg, Kurt A.; Bostrom, Meredith A.; Cooke, Jessica N.; Suzuki, Daisuke; Babazono, Tetsuya; Uzu, Takashi; Tang, Sydney C.W.; Mondal, Ashis; Sharma, Neeraj Kumar; Kobes, Sayuko; Antinozzi, Peter A.; Davis, Matthew; Das, Swapan K.; Rasouli, Neda; Kern, Philip A.; Shores, Nathan J.; Rudel, Lawrence L.; Blüher, Matthias; Stümvoll, Michael; Bowden, Donald W.; Maeda, Shiro; Parks, John S.; Kovacs, Peter; Hanson, Robert L.; Baier, Leslie J.; Elbein, Steven C.; Freedman, Barry I.

doi:10.1371/journal.pone.0056193

Cited by 13 publications

(10 citation statements)

References 32 publications

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“…11697498001)). 100 mg protein was loaded into individual wells of a Mini-PROTEAN TGX Precast Gel (Bio-Rad) and a western blot was performed as described [ 59 ]. Blots were incubated overnight at 4°C with anti-APOL1 antibody (1:1000; Abcam, EPR2907, ab108315).…”

Section: Methodsmentioning

confidence: 99%

In vivo Modeling Implicates APOL1 in Nephropathy: Evidence for Dominant Negative Effects and Epistasis under Anemic Stress

et al. 2015

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African Americans have a disproportionate risk for developing nephropathy. This disparity has been attributed to coding variants (G1 and G2) in apolipoprotein L1 (APOL1); however, there is little functional evidence supporting the role of this protein in renal function. Here, we combined genetics and in vivo modeling to examine the role of apol1 in glomerular development and pronephric filtration and to test the pathogenic potential of APOL1 G1 and G2. Translational suppression or CRISPR/Cas9 genome editing of apol1 in zebrafish embryos results in podocyte loss and glomerular filtration defects. Complementation of apol1 morphants with wild-type human APOL1 mRNA rescues these defects. However, the APOL1 G1 risk allele does not ameliorate defects caused by apol1 suppression and the pathogenicity is conferred by the cis effect of both individual variants of the G1 risk haplotype (I384M/S342G). In vivo complementation studies of the G2 risk allele also indicate that the variant is deleterious to protein function. Moreover, APOL1 G2, but not G1, expression alone promotes developmental kidney defects, suggesting a possible dominant-negative effect of the altered protein. In sickle cell disease (SCD) patients, we reported previously a genetic interaction between APOL1 and MYH9. Testing this interaction in vivo by co-suppressing both transcripts yielded no additive effects. However, upon genetic or chemical induction of anemia, we observed a significantly exacerbated nephropathy phenotype. Furthermore, concordant with the genetic interaction observed in SCD patients, APOL1 G2 reduces myh9 expression in vivo, suggesting a possible interaction between the altered APOL1 and myh9. Our data indicate a critical role for APOL1 in renal function that is compromised by nephropathy-risk encoding variants. Moreover, our interaction studies indicate that the MYH9 locus is also relevant to the phenotype in a stressed microenvironment and suggest that consideration of the context-dependent functions of both proteins will be required to develop therapeutic paradigms.

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Section: Methodsmentioning

confidence: 99%

In vivo Modeling Implicates APOL1 in Nephropathy: Evidence for Dominant Negative Effects and Epistasis under Anemic Stress

et al. 2015

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“…The upregulated genes included ACACB and NFKBIA. ACACB (acetyl-CoA carboxylase beta) is a rate-limiting enzyme involved in mitochondrial fatty acid oxidation and plays a key role in fatty acid metabolism [33]. NFKBIA (NFKB inhibitor alpha) is involved in inflammatory responses and is upregulated in AD [34].…”

Section: Molecular Biologymentioning

confidence: 99%

Microarray Analysis of Differential Gene Expression in Alzheimer’s Disease Identifies Potential Biomarkers with Diagnostic Value

Liu

Zhong

et al. 2020

Med Sci Monit

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Funding was provided by the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (No. 17KJB360003) Background: Alzheimer disease (AD) is a common and fatal subtype of dementia that remains a challenge to diagnose and treat. This study aimed to identify potential biomarkers that influence the prognosis of AD. Material/Methods: A total of 6 gene expression profiles from the Gene Expression Omnibus (GEO) database were assessed for their potential as AD biomarkers. We identified differentially expressed genes (DEGs) using the prediction analysis for microarray (PAM) algorithm and obtained hub genes through the analysis of the protein-protein interaction (PPI) network and module analysis. Results: We identified 6 gene expression profiles from the GEO database and assessed their potential as AD biomarkers. Shared gene sets were extracted and integrated into large expression profile matrices. We identified 2514 DEGs including 68 upregulated-and 2446 downregulated genes through analysis of the limma package. We screened 379 significant DEGs including 68 upregulated and 307 downregulated genes for their ability to distinguish AD from control samples using PAM algorithm. Functional enrichment of the 379 target genes was produced from Database for Annotation, Visualization and Integrated Discovery.(DAVID) and included histone function, beta receptor signaling, cell growth, and angiogenesis. The downregulated genes were significantly enriched in MAPK signaling, synaptic signaling, neuronal apoptosis and AD associated pathways. Upon analysis of the PPI network, 32 hub genes including ENO2, CCT2, CALM2, ACACB, ATP5B, MDH1, and PP2CA were screened. Of these hub genes, NFKBIA and ACACB were upregulated and 29 genes were downregulated in AD patients. Conclusions: We screened 379 significant DEGs as potential biomarkers of AD using PAM and obtained 32 hub genes through PPI network and module analysis. These findings reveal new potential AD biomarkers with prognostic and therapeutic value.

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“…33 Blots were incubated overnight at 4°C with anti-APOL1 antibody (1:1,000, HPA018885; Sigma-Aldrich). The membranes were then washed three times in Tris-buffered saline containing 0.1% Tween 20 and incubated for 1 hour in blocking buffer with anti-rabbit IgG conjugated to horseradish peroxidase (1:20,000; Jackson ImmunoResearch Laboratories, West Grove, PA).…”

Section: Western Blotmentioning

confidence: 99%

Localization of APOL1 Protein and mRNA in the Human Kidney

Ma¹,

Shelness²,

Snipes³

et al. 2015

Journal of the American Society of Nephrology

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118

115

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Although APOL1 gene variants are associated with nephropathy in African Americans, little is known about APOL1 protein synthesis, uptake, and localization in kidney cells. To address these questions, we examined APOL1 protein and mRNA localization in human kidney and human kidney-derived cell lines. Indirect immunofluorescence microscopy performed on nondiseased nephrectomy cryosections from persons with normal kidney function revealed that APOL1 protein was markedly enriched in podocytes (colocalized with synaptopodin and Wilms' tumor suppressor) and present in lower abundance in renal tubule cells. Fluorescence in situ hybridization detected APOL1 mRNA in glomeruli (podocytes and endothelial cells) and tubules, consistent with endogenous synthesis in these cell types. When these analyses were extended to renal-derived cell lines, quantitative RT-PCR did not detect APOL1 mRNA in human mesangial cells; however, abundant levels of APOL1 mRNA were observed in proximal tubule cells and glomerular endothelial cells, with lower expression in podocytes. Western blot analysis revealed corresponding levels of APOL1 protein in these cell lines. To explain the apparent discrepancy between the marked abundance of APOL1 protein in kidney podocytes observed in cryosections versus the lesser abundance in podocyte cell lines, we explored APOL1 cellular uptake. APOL1 protein was taken up readily by human podocytes in vitro but was not taken up efficiently by mesangial cells, glomerular endothelial cells, or proximal tubule cells. We hypothesize that the higher levels of APOL1 protein in human cryosectioned podocytes may reflect both endogenous protein synthesis and APOL1 uptake from the circulation or glomerular filtrate.

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An ACACB Variant Implicated in Diabetic Nephropathy Associates with Body Mass Index and Gene Expression in Obese Subjects

Cited by 13 publications

References 32 publications

In vivo Modeling Implicates APOL1 in Nephropathy: Evidence for Dominant Negative Effects and Epistasis under Anemic Stress

In vivo Modeling Implicates APOL1 in Nephropathy: Evidence for Dominant Negative Effects and Epistasis under Anemic Stress

Microarray Analysis of Differential Gene Expression in Alzheimer’s Disease Identifies Potential Biomarkers with Diagnostic Value

Localization of APOL1 Protein and mRNA in the Human Kidney

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