Obesity and type 2 diabetes are associated with disturbances in insulin-regulated glucose and lipid fluxes and severe comorbidities including cardiovascular disease and steatohepatitis. Whole body metabolism is regulated by lipid-storing white adipocytes as well as "brown" and "brite/beige" adipocytes that express thermogenic uncoupling protein 1 (UCP1) and secrete factors favorable to metabolic health. Implantation of brown fat into obese mice improves glucose tolerance, but translation to humans has been stymied by low abundance of primary human beige adipocytes. Here we apply methods to greatly expand human adipocyte progenitors from small samples of human subcutaneous adipose tissue and then disrupt the thermogenic suppressor gene NRIP1 by CRISPR. Ribonucleoprotein consisting of Cas9 and sgRNA delivered ex vivo are fully degraded by the human cells following high efficiency NRIP1 depletion without detectable off-target editing. Implantation of such CRISPR-enhanced human or mouse brown-like adipocytes into high fat diet fed mice decreases adiposity and liver triglycerides while enhancing glucose tolerance compared to implantation with unmodified adipocytes. These findings advance a therapeutic strategy to improve metabolic homeostasis through CRISPR-based genetic enhancement of human adipocytes without exposing the recipient to immunogenic Cas9 or delivery vectors.
- MeSH
- bílé tukové buňky metabolismus MeSH
- buněčná diferenciace MeSH
- buněčné kultury metody MeSH
- CRISPR-Cas systémy genetika MeSH
- dieta s vysokým obsahem tuků škodlivé účinky MeSH
- dospělé kmenové buňky fyziologie MeSH
- editace genu metody MeSH
- guide RNA, Kinetoplastida genetika MeSH
- hnědé tukové buňky metabolismus transplantace MeSH
- lidé MeSH
- metabolismus lipidů genetika MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- nuclear receptor interacting protein 1 genetika metabolismus MeSH
- obezita komplikace metabolismus terapie MeSH
- podkožní tuk cytologie MeSH
- porucha glukózové tolerance etiologie metabolismus terapie MeSH
- termogeneze genetika MeSH
- ztučnělá játra etiologie metabolismus prevence a kontrola MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
BACKGROUND: The de novo biosynthesis of fatty acids (DNL) through fatty acid synthase (FASN) in adipocytes is exquisitely regulated by nutrients, hormones, fasting, and obesity in mice and humans. However, the functions of DNL in adipocyte biology and in the regulation of systemic glucose homeostasis are not fully understood. METHODS & RESULTS: Here we show adipocyte DNL controls crosstalk to localized sympathetic neurons that mediate expansion of beige/brite adipocytes within inguinal white adipose tissue (iWAT). Induced deletion of FASN in white and brown adipocytes of mature mice (iAdFASNKO mice) enhanced glucose tolerance, UCP1 expression, and cAMP signaling in iWAT. Consistent with induction of adipose sympathetic nerve activity, iAdFASNKO mice displayed markedly increased neuronal tyrosine hydroxylase (TH) and neuropeptide Y (NPY) content in iWAT. In contrast, brown adipose tissue (BAT) of iAdFASNKO mice showed no increase in TH or NPY, nor did FASN deletion selectively in brown adipocytes (UCP1-FASNKO mice) cause these effects in iWAT. CONCLUSIONS: These results demonstrate that downregulation of fatty acid synthesis via FASN depletion in white adipocytes of mature mice can stimulate neuronal signaling to control thermogenic programming in iWAT.
- MeSH
- AMP cyklický metabolismus MeSH
- krevní glukóza metabolismus MeSH
- kultivované buňky MeSH
- lipogeneze * MeSH
- mastné kyseliny biosyntéza MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- neurony metabolismus fyziologie MeSH
- neuropeptid Y metabolismus MeSH
- sympatický nervový systém cytologie fyziologie MeSH
- syntázy mastných kyselin metabolismus MeSH
- termogeneze * MeSH
- tukové buňky metabolismus MeSH
- tyrosin-3-monooxygenasa metabolismus MeSH
- uncoupling protein 1 metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Research Support, N.I.H., Extramural MeSH
