Search

Pancreatic ductal adenocarcinoma (PDAC) is one of the top five deadliest forms of cancer with very few treatment options. The 5-year survival rate for PDAC is 10% following diagnosis. Preclinical murine models have been developed that leverage key driver mutations and have significantly contributed to our understanding of PDAC. One such genetically engineered mouse model (GEMM) that has emerged as an important tool is the KPC mouse (LSL-KrasG12D/+;LSL-Trp53R172H/+; p48-Cre) that spontaneously develops pancreatic tumors at ~14-16 weeks of age. Cadherin-11 (Cdh11), a cell-to-cell adhesion molecule has been suggested to play a role in development of the desmoplastic stroma in PDAC, that leads to difficulties in drug accessibility and has been hypothesized to contribute to chemotherapeutic resistance and correlate with poor prognosis. However, the mechanisms by which Cdh11 deficiency in the stromal microenvironment of PDAC-bearing KPC mice influences tumor infiltrating immune cells, has yet to be fully understood. Single-cell RNA sequencing (scRNAseq) of the immune (CD45+) compartment of tumor bearing Cdh11 proficient (KPC/Cdh11+/+), tumor bearing Cdh11 deficient (KPC/Cdh11+/-), non-tumor bearing Cdh11 deficient (Cdh11+/-) and wildtype (Cdh11+/+) mice was performed. We observed a sharp decrease in the presence of myeloid/monocyte lineage cells (CD14+) in KPC/Cdh11+/- tumors and also an increase in T, B and plasma cells, compared to KPC/Cdh11+/+ tumors. Genes upregulated in infiltrating T- and NK cells specific to a Cdh11 deficient background include Cd8a, Nkg7, Maf. Additionally, genes found to be upregulated in B cell clusters in Cdh11 deficient mice include those related to B cell differentiation/activation such as Lgals1, Id2, Itgb1, Rgs1. The increase in B and T cell infiltration was specific to the Cdh11 deficient background, since both pancreata from KPC/Cdh11+/+and Cdh11+/- mice had elevated levels of infiltration. Immunohistochemical validation of these findings has confirmed these changes in tumor infiltrating immune cells. Additionally of note, an increase in antibody-producing plasma cells was observed specifically in a Cdh11 deficient background. Igkc, an immunoglobulin found to be enriched in plasma cells was highly expressed in this group of immune cells. We also observed that KPC/Cdh11+/-had significantly more Igkc expressing cells than KPC/Cdh11+/+. Future work is needed to clearly define the role of Cdh11 in modulating B, T and plasma cell behavior and subsequent contributions to PDAC outcome. This study received funding by LDRD 19-SI-003. This work was conducted under the auspices of the USDOE by LLNL (DE-AC52-07NA27344), LLNL-ABS-820889. This work was supported by AACR-AstraZeneca Fellowship in Immunooncology Research, grant 17-40-12-PERA; The Ruesch Center for the Cure of Gastrointestinal Cancers grant award; NIH R01 CA170653; and NIH Cancer Center Support Grant P30 CA051008. Citation Format: Kelly A. Martin, Aimy Sebastian, Nicholas Hum, Ivana Peran, Stephen Byers, Elizabeth Wheeler, Matthew Coleman, Gabriela Loots. Loss of cadherin 11 in pancreatic cancer induces altered immune cell infiltration [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2516.

Introduction: Emerging evidence suggests that tumor extracellular matrix (ECM) may play a role in tumor-immune interactions. Breast tumors with high immune infiltrates have a distinct ECM profile, and T cell exclusion has been linked to specific ECM signatures. Despite this evidence suggesting a link between immune infiltrates and tumor matrix, it remains unclear whether ECM can directly affect the ultimate step in tumor clearance by the immune system, T cell mediated cytotoxicity. Methods: We compared clearance of 4T1 mammary gland carcinoma cells (MCC) seeded on ECM arrays by T cells isolated from spleens of MHC mismatched strain of mice. Briefly, 4T1 were seeded at 10k/ml for 1 hour, cultured for 24 hours then cocultured with T cells for 2 hours before fixing and staining. For RNA sequencing, ECM proteins (Collagen 1 -Col1, Collagen 4- Col4, Fibronectin -Fn or Vitronectin- Vtn) were coated onto plates at 250ug/ml, then 4t1 were added for 24 hours, then T cells were added for 24 hours followed by RNA isolation and sequencing. Results: We compared number of cells per spot with and without T cells across all ECM combinations and found that co-culture with T cells reduced the average number of MCCs, but this difference did not reach statistical significance. Only in the following conditions did MCC number significantly decrease: Col1 alone, Col6 alone, Fn alone, Vtn alone and Col6+ Eln (Fig 2B). In Col4 containing conditions, MCC cell number increased in the presence of T cells. Intensity of CD274 (PD-L1) and the MHC class 1 protein H2-Kd varied with substrate (p This work was funded by LDRD 19-SI-003 under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under contract DE-AC52-07NA27344. LLNL-ABS-8235222 Citation Format: Claire Robertson, Aimy Sebastian, Aubree Hinckley, Naiomy Rios-Arce, William Hynes, Wei He, Nicholas Hum, Elizabeth Wheeler, Gabriela Loots, Matthew Coleman, Monica Moya. Extracellular matrix modulates T cell clearance of malignant cells in vitro [abstract]. In: Abstracts: AACR Virtual Special Conference: Tumor Immunology and Immunotherapy; 2021 Oct 5-6. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(1 Suppl):Abstract nr P043.

Rift Valley Fever Virus (RVFV) is a category A biodefense pathogen due to the potential for intentional spread by aerosol and the lack of any licensed vaccine or antiviral therapeutics. Despite the brain being a major target of viral replication and tissue damage following aerosol infection, little is known about innate immune responses to RVFV in the brain. Thus, we are investigating the responses of resident innate immune cells of the brain to RVFV infection. Microglia and astrocytes mount a robust response to RVFV infection that is dependent upon MAVS and independent of TLR3 and TLR7. MAVS KO mice have significantly higher viral titers in the brain following intranasal infection, but surprisingly many markers of immune activity were similarly upregulated in the brains of WT and KO mice, including infiltration of CD45+ cells, upregulation of inflammatory genes, and cytokine expression. Indeed, many inflammatory and cytokine genes were upregulated more significantly in infected KO brains than in WT. This is in contrast to the in vitro response of microglia and astrocytes, where MAVS KO cells were virtually unresponsive to RVFV infection. RNA-seq analysis of infected brain tissue revealed that key signal transduction pathways are not activated in MAVS KO animals. These results are an important step towards understanding the precise molecular pathways responsible for controlling RVFV infection in the brain, and will be critical to informing the development of vaccines and antiviral therapeutics that are successful in preventing encephalitis caused by RVFV.