TY - JOUR
T1 - Disruption of FDPS/Rac1 axis radiosensitizes pancreatic ductal adenocarcinoma by attenuating DNA damage response and immunosuppressive signalling
AU - Seshacharyulu, Parthasarathy
AU - Halder, Sushanta
AU - Nimmakayala, Ramakrishna
AU - Rachagani, Satyanarayana
AU - Chaudhary, Sanjib
AU - Atri, Pranita
AU - Chirravuri-Venkata, Ramakanth
AU - Ouellette, Michel M.
AU - Carmicheal, Joseph
AU - Gautam, Shailendra K.
AU - Vengoji, Raghupathy
AU - Wang, Shuo
AU - Li, Sicong
AU - Smith, Lynette
AU - Talmon, Geoffrey A.
AU - Klute, Kelsey
AU - Ly, Quan
AU - Reames, Bradley N.
AU - Grem, Jean L.
AU - Berim, Lyudmyla
AU - Padussis, James C.
AU - Kaur, Sukhwinder
AU - Kumar, Sushil
AU - Ponnusamy, Moorthy P.
AU - Jain, Maneesh
AU - Lin, Chi
AU - Batra, Surinder K.
N1 - Funding Information:
Study design: PS, CL, and SKB, Experiment performed and data analysis: PS, SH, RN, SR, SC, PA, RCV, RV, JC, SKG, SW, SL, and GAT, Funding acquisition: MJ and SKB, Clinical investigation: KK, Q Ly, BNR, JLG, LB, JCP, and CL, Data verification: PS, LS, JC, PA, RCV, MPP, S Kaur, SK, MJ, CL, and SKB, Supervision: CL and SKB, Writing draft manuscript: PS, JC, and LS, Manuscript revision and final proofed: SH, RN, SR, SC, PA, RCV, MMO, JC, SKG, RV, SW, SL, LS, GAT, KK, Q Ly, BNR, JLG, LB, JCP, S Kaur, SK, MPP, MJ, CL, and SKB. All authors are aware of the data in this paper's content, read and approved the final version of the manuscript. The processed transcriptomic data, BAM and FASTQ files, and corresponding metadata are submitted to NCBI's SRA database (Bio project PRJNA739147, Submission-ID: SUB9857194). The work in this article was supported, in parts, by the following grants from the National Institutes of Health (P50 CA 127297 (Dr. Batra), P01 CA217798 (Dr. Batra), R01 CA195586 (Dr. Batra and Dr. Jain), R01 CA247471 (Dr. Batra and Dr. Jain), R01 CA210637 (Dr. Ponnusamy), and U01 CA213862 (Dr. Jain). We thank Kavita Mallya and Corinn E. Grabow for their technical help. We also thank Amy Filler-Katz and Erin L Doherty for their coordination in a clinical study. We thank Jessica Mercer for the editorial assistance of this manuscript. We thank the UNMC Sequencing Core facility for RNA seq. analysis in clinical specimens. We thank Craig Semerad and Victoria B. Smith of the Flow Cytometry Research Facility, UNMC, for assisting with flow cytometry. Schematic figure and cartoons were created using BioRender.com. Finally, we thank Janice A. Taylor and James R. Talaska of the Advanced Microscopy Core Facility at the UNMC for help with confocal microscopy.
Funding Information:
The work in this article was supported, in parts, by the following grants from the National Institutes of Health (P50 CA 127297 (Dr. Batra), P01 CA217798 (Dr. Batra), R01 CA195586 (Dr. Batra and Dr. Jain), R01 CA247471 (Dr. Batra and Dr. Jain), R01 CA210637 (Dr. Ponnusamy), and U01 CA213862 (Dr. Jain).
Publisher Copyright:
© 2021 The Author(s)
PY - 2022/1
Y1 - 2022/1
N2 - Background: Radiation therapy (RT) has a suboptimal effect in patients with pancreatic ductal adenocarcinoma (PDAC) due to intrinsic and acquired radioresistance (RR). Comprehensive bioinformatics and microarray analysis revealed that cholesterol biosynthesis (CBS) is involved in the RR of PDAC. We now tested the inhibition of the CBS pathway enzyme, farnesyl diphosphate synthase (FDPS), by zoledronic acid (Zol) to enhance radiation and activate immune cells. Methods: We investigated the role of FDPS in PDAC RR using the following methods: in vitro cell-based assay, immunohistochemistry, immunofluorescence, immunoblot, cell-based cholesterol assay, RNA sequencing, tumouroids (KPC-murine and PDAC patient-derived), orthotopic models, and PDAC patient's clinical study. Findings: FDPS overexpression in PDAC tissues and cells (P < 0.01 and P < 0.05) is associated with poor RT response and survival (P = 0.024). CRISPR/Cas9 and pharmacological inhibition (Zol) of FDPS in human and mouse syngeneic PDAC cells in conjunction with RT conferred higher PDAC radiosensitivity in vitro (P < 0.05, P < 0.01, and P < 0.001) and in vivo (P < 0.05). Interestingly, murine (P = 0.01) and human (P = 0.0159) tumouroids treated with Zol+RT showed a significant growth reduction. Mechanistically, RNA-Seq analysis of the PDAC xenografts and patients-PBMCs revealed that Zol exerts radiosensitization by affecting Rac1 and Rho prenylation, thereby modulating DNA damage and radiation response signalling along with improved systemic immune cells activation. An ongoing phase I/II trial (NCT03073785) showed improved failure-free survival (FFS), enhanced immune cell activation, and decreased microenvironment-related genes upon Zol+RT treatment. Interpretation: Our findings suggest that FDPS is a novel radiosensitization target for PDAC therapy. This study also provides a rationale to utilize Zol as a potential radiosensitizer and as an immunomodulator in PDAC and other cancers. Funding: National Institutes of Health (P50, P01, and R01).
AB - Background: Radiation therapy (RT) has a suboptimal effect in patients with pancreatic ductal adenocarcinoma (PDAC) due to intrinsic and acquired radioresistance (RR). Comprehensive bioinformatics and microarray analysis revealed that cholesterol biosynthesis (CBS) is involved in the RR of PDAC. We now tested the inhibition of the CBS pathway enzyme, farnesyl diphosphate synthase (FDPS), by zoledronic acid (Zol) to enhance radiation and activate immune cells. Methods: We investigated the role of FDPS in PDAC RR using the following methods: in vitro cell-based assay, immunohistochemistry, immunofluorescence, immunoblot, cell-based cholesterol assay, RNA sequencing, tumouroids (KPC-murine and PDAC patient-derived), orthotopic models, and PDAC patient's clinical study. Findings: FDPS overexpression in PDAC tissues and cells (P < 0.01 and P < 0.05) is associated with poor RT response and survival (P = 0.024). CRISPR/Cas9 and pharmacological inhibition (Zol) of FDPS in human and mouse syngeneic PDAC cells in conjunction with RT conferred higher PDAC radiosensitivity in vitro (P < 0.05, P < 0.01, and P < 0.001) and in vivo (P < 0.05). Interestingly, murine (P = 0.01) and human (P = 0.0159) tumouroids treated with Zol+RT showed a significant growth reduction. Mechanistically, RNA-Seq analysis of the PDAC xenografts and patients-PBMCs revealed that Zol exerts radiosensitization by affecting Rac1 and Rho prenylation, thereby modulating DNA damage and radiation response signalling along with improved systemic immune cells activation. An ongoing phase I/II trial (NCT03073785) showed improved failure-free survival (FFS), enhanced immune cell activation, and decreased microenvironment-related genes upon Zol+RT treatment. Interpretation: Our findings suggest that FDPS is a novel radiosensitization target for PDAC therapy. This study also provides a rationale to utilize Zol as a potential radiosensitizer and as an immunomodulator in PDAC and other cancers. Funding: National Institutes of Health (P50, P01, and R01).
KW - FDPS
KW - PDAC
KW - Radioresistance
KW - Radiotherapy
KW - Zoledronic acid
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U2 - 10.1016/j.ebiom.2021.103772
DO - 10.1016/j.ebiom.2021.103772
M3 - Article
C2 - 34971971
AN - SCOPUS:85121972956
SN - 2352-3964
VL - 75
JO - EBioMedicine
JF - EBioMedicine
M1 - 103772
ER -