Engineered allele substitution at PPARGC1A rs8192678 alters human white adipocyte differentiation, lipogenesis, and PGC-1α content and turnover
Huang, Mi LU ; Claussnitzer, Melina ; Saadat, Alham ; Coral, Daniel LU
; Kalamajski, Sebastian
LU
and Franks, Paul
LU
(2023)
In Diabetologia
66(7).
p.1289-1305
- 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... (More) - 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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/40e5de83-3b54-40a4-a2a2-47fb15c425ec
- author
- Huang, Mi
LU
; Claussnitzer, Melina
; Saadat, Alham
; Coral, Daniel
LU
; Kalamajski, Sebastian
LU
and Franks, Paul
LU
- organization
- publishing date
- 2023-05-12
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Gene and environment interaction, GWAS, Obesity, PGC-1a, PPARGC1A, rs8192678
- in
- Diabetologia
- volume
- 66
- issue
- 7
- pages
- 17 pages
- publisher
- Springer
- external identifiers
-
- pmid:37171500
- scopus:85159270950
- ISSN
- 1432-0428
- DOI
- 10.1007/s00125-023-05915-6
- language
- English
- LU publication?
- yes
- id
- 40e5de83-3b54-40a4-a2a2-47fb15c425ec
- date added to LUP
- 2023-05-12 14:42:31
- date last changed
- 2023-10-26 14:48:11
@article{40e5de83-3b54-40a4-a2a2-47fb15c425ec,
abstract = {{Aims/hypothesis<br/><br/>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.<br/>Methods<br/><br/>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.<br/>Results<br/><br/>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.<br/>Conclusions/interpretation<br/><br/>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.}},
author = {{Huang, Mi and Claussnitzer, Melina and Saadat, Alham and Coral, Daniel and Kalamajski, Sebastian and Franks, Paul}},
issn = {{1432-0428}},
keywords = {{Gene and environment interaction; GWAS; Obesity; PGC-1a; PPARGC1A; rs8192678}},
language = {{eng}},
month = {{05}},
number = {{7}},
pages = {{1289--1305}},
publisher = {{Springer}},
series = {{Diabetologia}},
title = {{Engineered allele substitution at PPARGC1A rs8192678 alters human white adipocyte differentiation, lipogenesis, and PGC-1α content and turnover}},
url = {{http://dx.doi.org/10.1007/s00125-023-05915-6}},
doi = {{10.1007/s00125-023-05915-6}},
volume = {{66}},
year = {{2023}},
}