Ablation of hematopoietic stem cell derived adipocytes reduces tumor burden in syngeneic mouse models of high-grade serous carcinoma
Authors: Woodruff Elizabeth R , Bailey Courtney A , To Francis , Manda Vyshnavi , Maltzahn Joanne K , Sullivan Timothy M , Boorgula Meher P , Recouvreux Maria Sol , Vianzon Ruby , Conrad Bogi , Gavin Kathleen M , Jordan Kimberly R , Klemm Dwight J , Orsulic Sandra , Bitler Benjamin G , Watson Zachary L 
Published: September 2024

ABSTRACTIn this study we examined the influence of hematopoietic stem cell-derived adipocytes (HSCDAs) on the proliferation and metastasis of high-grade serous carcinoma (HGSC) – the most common type of ovarian cancer. HSCDAs are a subtype of adipocytes that differentiate from myeloid precursors that traffic from bone marrow to adipose tissue and accumulate therein. These are distinct from conventional mesenchymal adipocytes (CMAs), which are derived from mesenchymal precursors. We hypothesized that HSCDAs promote HGSC progression and establish a pro-tumoral niche within peritoneal adipose tissues such as the omentum.Primary human white adipose tissue samples were obtained via biopsy and then sorted into myeloid and mesenchymal populations through flow cytometry. These adipose precursors were then differentiatedin vitrointo mature HSCDAs and CMAs, respectively. Transcriptomic analysis showed that HSCDAs have a distinct transcriptional profile from CMAs, including downregulation of cell cycle and upregulation of multiple metabolic and adipogenic pathways. Using ELISA, we found that HSCDAs secreted greater amounts of inflammatory cytokines IL-6 and IL-8 than CMAs. Next, we incubated HGSC cells in conditioned media from HSCDAs and CMAs and performed proliferation and protein expression profiling. HGSC cells in HSCDA media, compared to those in CMA media, had elevated expression of protein markers related to epithelial to mesenchymal plasticity, including fibronectin, as well as increased serine phosphorylation of pro-survival AKT1/2. Conversely, HGSC cells in HSCDA media exhibited comparably downregulated expression of tumor suppressors including the Wnt regulator GSK3β. Depending on the cell line and adipose donor, HGSC cells also showed altered growth rates in conditioned media.We next investigated the role of HSCDAs in HGSC progression and metastasisin vivo. We generated immunocompetent mice that were either HSCDA Proficient (can make both adipocyte subtypes) or Deficient (can only make CMAs). Using these models, we conducted two independent tumor studies using the ID8 (Tp53-/-,Brca2-/-) and SO (Tp53-/-,Brca1/2wild-type,HrasandMycamplified) syngeneic models. Overall tumor burden was lower in HSCDA Deficient mice in both models. In the ID8 model, omental tumors from HSCDA Deficient mice showed reduced proliferation (Ki67) and apoptosis (cleaved caspase 3) relative to those from Proficient mice.Transcriptionally, omental ID8 tumors from HSCDA Deficient downregulated oxidative phosphorylation, adipogenesis, and fatty acid metabolism relative to tumors from HSCDA Proficient mice. These pathways were enriched in HSCDA cellsin vitro, suggesting that ablation of HSCDAs had a significant influence on the tumor metabolic environment. Reduced inflammatory pathways in ID8 tumors from HSCDA Deficient mice were also observed leading us to interrogate immune cell infiltration into omental tumors. Compared to HSCDA Proficient mice, tumors from HSCDA Deficient mice showed reduced densities of dendritic cells (DC) and natural killer (NK) cells, as well as fewer DCs, NKs, and B-cells in proximity to tumor cells, as determined by spatial analysis. Overall, our data suggest that HSCDAs promote HGSC survival and plasticity while downregulating expression of tumor suppressors and altering the peritoneal immune and metabolic environment to promote HGSC progression.