Pulsed Low-Dose-Rate Radiation (Pldr) Limits Pancreatic Pro-Tumor Stroma Aggravation: Pre-Clinical Basis For An Ongoing Pldr Trail.

Abstract Radiation, combined with radio-sensitizing chemotherapy, is used preoperatively as neoadjuvant therapy (NT) for borderline resectable pancreatic ductal adenocarcinoma (PDAC) with the intent of facilitating a curative surgical intervention. PDAC is uniquely characterized by an extensive fibrous microenvironment, desmoplasia, which can unintendedly be aggravated by NT to foster its pro-tumoral function. Radiation, as part of NT, is aimed at providing a margin adjacent to un-resectable vessels and sterilizing regional lymph nodes. Yet, to avoid the unintended desmoplastic aggravation, and because of a risk of toxicities associated with high doses due to the radio-sensitivity of adjacent small bowel and stomach, the total dose of NT radiation delivered to PDAC patients is classically modest. Therefore, NT radiation in PDAC patients is often lower than optimal for effective tumor cell elimination. Pulsed low-dose-rate (PLDR) radiation improves the safety of radiation treatment as it allows time for DNA damage repair in non-tumorous cells/tissues while simultaneously remaining as effective as continuous dose rate (CDR) radiation in cancer cells. In theory, PLDR could be amendable for increased radiation dosage. Of interest, the use of PLDR in pre-clinical animal studies revealed a systemic lowering of transforming growth factor beta (TGFβ), a known immunosuppressive factor. Hence, we posit that since cancer-associated fibroblasts (CAFs) produce TGFβ and constitute one of the most abundant cells in PDAC desmoplasia, variations in TGFβ levels in response to NT will inform on desmoplastic dynamic changes. Our team has developed means to assess CAF functions and activation statuses. These efforts were guided by generating a desmoplastic biomarker signature obtained from patient-harvested CAFs during the production of extracellular matrix (ECM), using our well-established in vitro 3D system. We employed this in vivo-mimetic system to test the hypothesis that PLDR limits pro-tumoral desmoplastic CAF aggravation. As part of the CAF functional signature, we tested levels of immunosuppressive TGFβ secretion, expression of netrin-G1, palladin, and others; together with the biogenesis of unique extracellular vesicles, and the production of ECMs capable of nurturing PDAC cells under starvation. During a 5-day assay, human PDAC 3D ECM producing CAFs were treated with gemcitabine (Gem; 5nm) plus 4Gy or 8Gy PLDR vs. CDR. Results indicated that Gem alone or with CDR indeed aggravated CAF’s pro-PDAC phenotype and function, while Gem combined with PDLR limited and sometimes reverted this functional pro-tumor CAF signature. Based on these results, we are poised to test this functional CAF/desmoplasia signature, as laboratory correlatives, in an ongoing clinical trial at Fox Chase Cancer Center. The phase I trial is a dose-escalation study of PLDR radiation and chemotherapy at standard and intensified doses in preoperative pancreatic cancer patients with toxicity, histopathologic, and CAF-informing translational endpoints. Citation Format: Janusz Franco-Barraza, Tiffany Luong, Jessica K. Wong, Debora B. Vendramini-Costa, Ralph Francescone, Jaye C. Gardiner, Kristopher S. Raghavan, Joshua E. Meyer, Edna Cukierman. Pulsed low-dose-rate radiation (PLDR) limits pancreatic pro-tumor stroma aggravation: Pre-clinical basis for an ongoing PLDR trail [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PR-006.