Bioengineering adipocytes for cancer therapy

PROJECT SUMMARY Increased glucose and fat metabolism are essential for tumor progression. Recent work showed that cold- induced activation of brown adipose tissue (BAT), a unique adipose tissue which produces heat via glycolysis and fat metabolism, suppressed tumor progression in several cancer mouse models due to competition for glucose and fat resources. White adipose tissue (WAT) is an endocrine tissue that functions as the main energy storage organ in our body, storing lipids and secreting hormones that regulate various biological functions, including appetite, glucose and fat metabolism, and insulin hemostasis. It is commonly used in clinical procedures such as liposuction and fat transplantation in plastic surgery. Thus, WAT could be readily used for cellular therapy. Here, we propose to showcase the utility of this approach, which we term as Adipose Manipulation Transplantation (AMT), for cancer therapy. We plan to bioengineer WAT and adipose organoids to become more BAT-like, which will increase their glucose and fat utilization, along with having increased glucose uptake and fat storage. This will be done using CRISPR activation (CRISPRa) to upregulated genes that are involved in BAT function and glucose and fat metabolism. Our preliminary results already show that co- culturing CRISPRa engineered BAT-like adipocytes with five different cancer cell lines suppresses their growth. Furthermore, implanting engineered adipose organoids with xenografts or in two cancer genetic mouse models (breast and pancreas) significantly reduces cancer growth. Finally, engineering adipocytes from eight different human breast cancer patients surgically resected tissue and co-culturing them with cancer organoids significantly reduced organoid growth. Here, we plan to build on these results. We will engineer adipocytes and organoids to have increased glucose and fat utilization by upregulating combinations of genes not only involved in browning, but also in glucose and fat metabolism and glucose transport and test their ability to reduce cancer growth in various cancer cell models (Aim 1). We will also test the ability of these CRISPRa gene combinations in adipose organoids to suppress cancer in xenograft cancer models from various cell lines (breast, colon, pancreas, prostate) and in several genetic mouse models (breast and pancreas) (Aim 2). Finally, to further dissect the translational potential of this approach, we will use surgically resected tumor tissue from breast cancer patients, isolate and engineer their adipocytes and then co-culture them with their respective tumor to assess their therapeutic potential (Aim 3). This will be done across the full spectrum of human breast cancer stages, including different stages of breast cancer (adjuvant, late recurrence, metastasis, challenging-to-treat breast cancer subtypes and BRCA mutations). Combined, this project will develop a novel ‘CAR T like’ therapeutic approach to treat cancer utilizing bioengineered adipocytes, having tremendous therapeutic implications for cancer treatment.