Abstract 3191: Tumor intrinsic HuR promotes stroma activation in pancreatic cancer
Authors: McCarthy Grace A. , Finan Jennifer M. , Jain Aditi , DiNiro Roberto , Grossberg Aaron , Brody Jonathan R. 
Published: June 2022

Background: One of the factors contributing to poor prognosis of pancreatic ductal adenocarcinoma (PDAC) is thought to be the characteristically dense stroma and heterogeneous tumor microenvironment. The PDAC stroma involves multiple players, including cancer associated fibroblasts (CAFs) and pancreatic stellate cells (PSCs). These stromal cells, when activated, promote extensive fibrosis in PDAC, which is associated with poor prognosis and chemoresistance.

We have previously established that the RNA-binding protein Human Antigen R (HuR, ELAVL1) facilitates PDAC progression. In a tumor microarray of matched normal and malignant patient tissues, we found high active (i.e. cytoplasmic) HuR staining in 80% of PDAC tumors, while low or absent in adjacent normal tissues (n=80). Moreover, in both the orthotopic and the genetically engineered mouse models of PDAC, active HuR staining in tumors was dramatically higher than in normal pancreas. Pancreas-specific overexpression of HuR in a transgenic mouse model did not initiate tumorigenesis, but did increase fibrosis compared to control mice. We hypothesize that tumor intrinsic HuR regulates transcripts that are needed for tumor cells to activate the surrounding stroma.

Methods: MiaPaCa-2 or PANC-1 human PDAC cells, either HuR-proficient or CRISPR-mediated knocked-out, were used in an orthotopic model where cells were injected into the tail of the pancreas. Both NRG and athymic mice were used. Resulting tumors underwent immunohistochemistry, immunofluorescence, or RNA-sequencing. Secreted proteins from cells in 2D culture were analyzed using a cytokine array.

Results: HuR knockout tumors showed a decrease in CAF and PSC markers (i.e. podoplanin, alpha-SMA, desmin; &gt;55% decrease, p&lt;0.05), as well as a decrease in collagen deposition (56% decrease, p=0.0004). Additionally, HuR expression in tumors positively correlated with collagen abundance (p= 0.0002).

RNA sequencing of PDAC tumors at various times during tumor development found that gene expression of stroma-activating ligands was significantly downregulated in HuR knockout tumors (e.g. TGFb, CCL2, PDGFA; -0.8&lt;log2FC&lt;-3.6, 0.0009&lt;padj&lt;2^-57). Moreover, we found a significant decrease of these stroma-activating ligands in conditioned media obtained from HuR knockout compared to wild type (e.g. PDGFAA, PTX3, CCL2; &gt;50% decrease, p&lt;0.0001).

Conclusions: We found inhibition of HuR led to a robust decrease in stroma activation (i.e. lack of collagen, CAFs, and PSCs). To our knowledge, this is the first report of PDAC tumor intrinsic HuR having an extrinsic effect on neighboring stromal cells. Specifically, we found loss of HuR results in a decrease of secreted stroma-activating ligands, likely to be the cause of the decreased fibrosis in HuR knockout tumors. With evidence that fibrosis is associated with poor survival rates and chemoresistance, inhibiting HuR may sensitize tumors to standard of care therapy.

Citation Format: Grace A. McCarthy, Jennifer M. Finan, Aditi Jain, Roberto DiNiro, Aaron Grossberg, Jonathan R. Brody. Tumor intrinsic HuR promotes stroma activation in pancreatic cancer [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 3191.