Oncogene-induced matrix reorganization controls CD8+ T cell function in the soft-tissue sarcoma microenvironment

Abstract

† Equally contributed. AMF and HCP can each list their name first on their respective CVs.

Collagen modification remodels the sarcoma tumor microenvironment and promotes resistance to immune checkpoint inhibition

“…These animals lack mature adaptive immune cells (T cells and B cells) but generally retain innate immunity (e.g., myeloid-lineage cells). Therefore, to understand the role of UPS cell-intrinsic Plod2 in immune evasion, we leveraged the Kras G12D/+ ; Trp53 fl/fl (KP) syngeneic transplant system previously introduced in 16 . In this system, sarcoma cells derived from the gold-standard KP genetically engineered mouse model (GEMM) of UPS 29 on a pure C57BL/6 background (herein referred to as “KP cells”) are implanted subcutaneously or orthotopically (intramuscularly) into syngeneic, immunocompetent C57BL/6 mice.…”

“…The unstained tissue slides were maintained in water, and image stacks (6.99 μm) were acquired using a 25X 1.0NA water immersion objective with 4x zoom. Collagen fiber width and orientation distribution from SHG image stacks (maximum-intensity projections) were quantified using CT-FIRE as described previously 16, 30 .…”

“…The xCELLigence real-time cytotoxicity assay was performed as previously described. 16 In brief, STS109 cells (target cells) expressing a control or one of multiple PLOD2 -targeting shRNAs were harvested and seeded in triplicate into 96-well polyethylene terephthalate plates (E-Plate VIEW 96 PET, Agilent) at 20,000 cells/per well. STS-109 cells were allowed to adhere to the bottoms of wells for 24 hours (37°C, 5% CO 2 conditions) after which effector cells (CART-TnMUC1 cells or donor-matched un-transduced T cells [NTD cells]) were added to wells at effector:target cell ratios of 10:1, 5:1, 1:1, and 0:1.…”

“…In particular, the solid tumor ECM is rich in molecules belonging to the collagen superfamily, a large, diverse protein family that contains 28 molecular species encoded by over 40 genes 15 . Although the roles of individual collagen proteins in cancer-associated processes are only beginning to be defined - particularly with respect to their impact on solid tumor immune evasion - our recent work demonstrated that collagen type VI (ColVI), a microfibrillar collagen, facilitates tumor progression by inhibiting T cell migration, infiltration, and function 16, 17 . Specifically, ColVI promotes immune evasion and hinders ICI efficacy in UPS by inactivating CD8 + T cells and remodeling collagen type I (ColI) in the TME 16 .…”

“…Although the roles of individual collagen proteins in cancer-associated processes are only beginning to be defined - particularly with respect to their impact on solid tumor immune evasion - our recent work demonstrated that collagen type VI (ColVI), a microfibrillar collagen, facilitates tumor progression by inhibiting T cell migration, infiltration, and function 16, 17 . Specifically, ColVI promotes immune evasion and hinders ICI efficacy in UPS by inactivating CD8 + T cells and remodeling collagen type I (ColI) in the TME 16 . These findings implicate ColVI ablation, in combination with immunotherapy, as a promising approach to mitigate immune evasion in UPS, and potentially other solid tumors.…”