Increased Fibrosis in White Adipose Tissue of Male and Female bGH Transgenic Mice Appears Independent of TGF-[beta] Action

Fibrosis is a pathological state caused by excess deposition of extracellular matrix proteins in a tissue. Male bovine growth hormone (bGH) transgenic mice experience metabolic dysfunction with a marked decrease in lifespan and with increased fibrosis in several tissues including white adipose tissue (WAT), which is more pronounced in the subcutaneous (Sc) depot. The current study expanded on these initial findings to evaluate WAT fibrosis in female bGH mice and the role of transforming growth factor (TGF)-[beta] in the development of WAT fibrosis. Our findings established that female bGH mice, like males, experience a depot-dependent increase in WAT fibrosis, and bGH mice of both sexes have elevated circulating levels of several markers of collagen turnover. Using various methods, TGF-[beta] signaling was found unchanged or decreased--as opposed to an expected increase--despite the marked fibrosis in WAT of bGH mice. However, acute GH treatments in vivo, in vitro, or ex vivo did elicit a modest increase in TGF-[beta] signaling in some experimental systems. Finally, single nucleus RNA sequencing confirmed no perturbation in TGF-[beta] or its receptor gene expression in any WAT cell subpopulations of Sc bGH WAT; however, a striking increase in B lymphocyte infiltration in bGH WAT was observed. Overall, these data suggest that bGH WAT fibrosis is independent of the action of TGF-[beta] and reveals an intriguing shift in immune cells in bGH WAT that should be further explored considering the increasing importance of B cell--mediated WAT fibrosis and pathology. Key Words: growth hormone, adipose tissue, fibrosis, TGF-[beta], bGH mice, pirfenidone Abbreviations: ANOVA, analysis of variance; bGH, bovine growth hormone; DMSO, dimethyl sulfoxide; FGF21, fibroblast growth factor 21; GH, growth hormone; ICTP, cross-linked C-terminal telopeptide of type I collagen; IGF, insulin-like growth factor; IL, interleukin; P1CP, procollagen I C-terminal propeptide; P1NP, procollagen I N-terminal propeptide; P3NP, procollagen III N-terminal propeptide; P3CP, procollagen III C-terminal propeptide; Peri, perigonadal; PGE2, prostaglandin E2; qPCR, quantitative polymerase chain reaction; Sc, subcutaneous; TGF, transforming growth factor; WAT, white adipose tissue.