Introduction and Objective: To achieve adipose mass reduction while maintaining or improving insulin sensitivity involves an integration of multiple neuroendocrine and paracrine inputs that requires an in vivo setting to be adequately studied. We therefore performed in vivo Perturb-Seq of visceral adipocytes to identify novel targets for the treatment of obesity.Methods: An AAV library encoding gRNA was administered to a Cas9 mouse model of obesity at a dose titrated to knockout a single gene per adipocyte. The transcriptomic responses to these perturbations were determined by single nucleus RNA-Sequencing of epididymal white adipose tissue and assessed according to curated ‘Disease Features’. A candidate perturbation was knocked down specifically in adipocytes as a single saturating gene therapy in obese mice to validate the therapeutic effect.Results: Despite the similar obesogenic and insulin sensitizing phenotypes described for the KO mice in the literature, Ncor1 KO and Pten KO adipocytes showed strongly divergent transcriptomic responses. Ncor1 KO adipocytes strongly upregulated features suggestive of metabolic health, including ‘Mitochondrial Biogenesis’, ‘Anaplerosis’, ‘Angiogenesis’ and multiple lipid synthesis features. Pten KO adipocytes downregulated these features, along with suppression of ‘Intracellular Lipolysis’. The insulin sensitizing effect of Pten deficiency was most evident in the strong upregulation of a ‘Cytoskeleton’ disease feature that has Cobll1 as a driver gene. Pten KO adipocytes also increased collagen gene expression, a finding that was validated using siRNA in ex vivo human adipocyte culture. A candidate perturbation with indications of increased lipolytic activity was found in a saturating experiment to decrease adipose mass while preserving lean mass and glucose tolerance.Conclusion: Our in vivo screening platform is capable of identifying novel therapeutic targets in an adipocyte type that is challenging to culture.
J. Hsieh: None. J. Chen: None. V. Kartha: None. L. Chio: None. D.R. Fuentes: None. C. Carrico: None. S. Hayward: None. I. Driver: Stock/Shareholder; Current; AbbVie Inc., Amgen Inc. C. Longtine: None. G. Pharaoh: None. A. Gupta: None. D. Popov: None. S. Shambhu: None. A. Lim: None. K. Doroschak: None. A. Vaidyanathan: None. J.M. Cruz Sampedro: None. S. Ruiz: None. J. Nonora: None. C.Q. Bowman: None. D. Shao: None. L. Fung: None. C. Towne: None. M.B. Jensen: None. F. LePort: None.
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