Transcriptomics of 3D model of adipose tissue

3D model to study the immunomodulatory functions of adipocyte-T lymphocytes paracrine crosstalkThe Adipose tissue (AT) is a critical endocrine and immunological organ that regulates systemic homeostasis. Obesity and type 2 diabetes, have an inflammatory component provided by both metabolic and immune cells. Among the immune cells, T lymphocytes play a critical role in driving AT inflammation and dysfunction by regulating macrophages and adipocytes. The interplay between adipocytes and T cells in the inflammatory response may rely on both direct cell-cell contact and cytokine production.Our first aim was to generate a model of AT-derived adipospheres from the subcutaneous biopsies of patients with different levels of metabolic perturbations. Following in vitro 3D differentiation of the stromal fraction cell bank into pre-vascularized adipospheres, we analyzed the effects of normocaloric and hypercaloric conditions on the morpho-functions of matured AT organoids. We demonstrated that adipocyte dysfunction and a pro-inflammatory state occurred upon metabolic cues in the absence of T cells. The adipokine secretory profile is influenced by donor-related factors, resulting in patient-specific responses in vitro.Subsequently, to decipher the crosstalk between adipocytes and T cells independently of systemic factors, we designed a new paracrine 3D co-culture composed of mature adipospheres in Transwell settings with blood-derived T cells. We provided a comparative analysis of single adipospheres cultures and co-cultures with T cells, encompassing adipokine, cytokine, lactate secretion profiles and metabolic phenotype. We also characterized the transcriptomic profiles of adipospheres. We showed that indirect interaction with T cells modulates the immune response of adipospheres towards inflammation. According to our model, we observed that dysfunctional subcutaneous AT adipocytes perturb T cell responses.This approach may be useful to (i) decipher the contribution of obesity to the loss of efficiency of immune responses, addressing both the profile of infected cells and effector immune cells, (ii) the interaction between myeloid and T cells within the AT environment, and (iii) the potential impact of drugs targeting metabolic pathways.