Engineered allele substitution at PPARGC1A rs8192678 alters human white adipocyte differentiation, lipogenesis, and PGC-1α content and turnover

Diabetologia · 6 authors, 3 centres

AI SUMMARY

FIDELITY 100%

POPULATIONIsogenic human white pre-adipocyte (hWAs) cell line

INTERVENTIONCRISPR-Cas9 allelic substitution at PPARGC1A rs8192678 generating homozygous C/C, T/T, and heterozygous C/T clones

COMPARISONHomozygous C/C vs T/T vs heterozygous C/T isogenic cell populations after adipogenic differentiation

This summary was generated by AI from a single paper. It has not been reviewed by a clinician and is not clinical advice. Verify against the source before acting on it.

AGENT 01 · STRUCTURED SUMMARY

TRACE TO SOURCE

“

TL;DR

Using CRISPR-Cas9 allelic switching at PPARGC1A rs8192678 in an isogenic human pre-adipocyte cell line, the authors show that the C allele (482Gly) reduces adipocyte differentiation, triacylglycerol accumulation, lipogenesis, and mitochondrial respiration compared with the T allele (482Ser) in a dose-dependent manner.

Full summary

957 CHARS

The authors used CRISPR-Cas9 genome editing to perform an allelic switch at PPARGC1A rs8192678 in an isogenic human white pre-adipocyte cell line, generating homozygous T/T, C/C, and heterozygous C/T clones. After adipogenic differentiation, C/C adipocytes showed visibly less BODIPY staining and significantly lower triacylglycerol content than T/T and C/T adipocytes. The C allele presented a dose-dependent lowering effect on lipogenesis, adipogenic gene expression, and oxygen consumption rate at basal and maximal respiration. These effects were attributed to C-allele-driven dysregulation of PGC-1α protein content, turnover rate, and transcriptional coactivator activity. The T allele conversely improved lipogenesis and triacylglycerol accumulation. Limitations include that experiments were performed in a single cell type from a single genetic background, and controlled gene-by-environment interaction experiments could not be performed in vitro.

PICO

PPOPULATION

Isogenic human white pre-adipocyte (hWAs) cell line

IINTERVENTION

CRISPR-Cas9 allelic substitution at PPARGC1A rs8192678 generating homozygous C/C, T/T, and heterozygous C/T clones

OOUTCOME

Adipocyte differentiation, triacylglycerol content, lipogenesis, gene expression, oxygen consumption rate, and PGC-1α protein content and turnover

STUDY TYPE

other

SPECIALTY

endocrinology

SUMMARISED BY

AI pipeline

FIDELITY CHECK

100% · A

ORIGINAL PAPER

PMC10244287

Engineered allele substitution at PPARGC1A rs8192678 alters human white adipocyte differentiation, lipogenesis, and PGC-1α content and turnover

Diabetologia

READ THE FULL PAPER

ABSTRACT

Aims/hypothesis PPARGC1A encodes peroxisome proliferator-activated receptor γ coactivator 1-α (PGC-1α), a central regulator of energy metabolism and mitochondrial function. A common polymorphism in PPARGC1A (rs8192678, C/T, Gly482Ser) has been associated with obesity and related metabolic disorders, but no published functional studies have investigated direct allele-specific effects in adipocyte biology. We examined whether rs8192678 is a causal variant and reveal its biological function in human white adipose cells. Methods We used CRISPR-Cas9 genome editing to perform an allelic switch (C-to-T or T-to-C) at rs8192678 in an isogenic human pre-adipocyte white adipose tissue (hWAs) cell line. Allele-edited single-cell clones were expanded and screened to obtain homozygous T/T (Ser482Ser), C/C (Gly482Gly) and heterozygous C/T (Gly482Ser) isogenic cell populations, followed by functional studies of the allele-dependent effects on white adipocyte differentiation and mitochondrial function. Results After differentiation, the C/C adipocytes were visibly less BODIPY-positive than T/T and C/T adipocytes, and had significantly lower triacylglycerol content. The C allele presented a dose-dependent lowering effect on lipogenesis, as well as lower expression of genes critical for adipogenesis, lipid catabolism, lipogenesis and lipolysis. Moreover, C/C adipocytes had decreased oxygen consumption rate (OCR) at basal and maximal respiration, and lower ATP-linked OCR. We determined that these effects were a consequence of a C-allele-driven dysregulation of PGC-1α protein content, turnover rate and transcriptional coactivator activity. Conclusions/interpretation Our data show allele-specific causal effects of the rs8192678 variant on adipogenic differentiation. The C allele confers lower levels of PPARGC1A mRNA and PGC-1α protein, as well as disrupted dynamics of PGC-1α turnover and activity, with downstream effects on cellular differentiation and mitochondrial function. Our study provides the first experimentally deduced insights on the effects of rs8192678 on adipocyte phenotype. Graphical Abstract Supplementary Information The online version of this article (10.1007/s00125-023-05915-6) contains peer-reviewed but unedited supplementary material.

CITATION EXPORT

Mi Huang, Melina Claussnitzer, Alham Saadat et al.. Engineered allele substitution at PPARGC1A rs8192678 alters human white adipocyte differentiation, lipogenesis, and PGC-1α content and turnover. Diabetologia. (2023). https://doi.org/10.1007/s00125-023-05915-6

AUTHORS

6 · 3 CENTRES

Mi Huang, Melina Claussnitzer, Alham Saadat, Daniel E. Coral, Sebastian Kalamajski, Paul W. Franks

1grid.4514.40000 0001 0930 2361Department of Clinical Sciences, Genetic and Molecular Epidemiology Unit, Clinical Research Centre, Lund University, Malmö, Sweden
2grid.66859.340000 0004 0546 1623Metabolism Program, Broad Institute of MIT and Harvard, Cambridge, MA 02142 USA
3grid.38142.3c000000041936754XDepartment of Nutrition, Harvard T.H. Chan School of Public Health, Boston, MA USA