Neutrophil-mediated fibroblast-tumor cell IL-6/STAT-3 signaling underlies the association between neutrophil-to-lymphocyte ratio dynamics and chemotherapy response in localized pancreatic cancer: a hybrid clinical-preclinical study

  1. Iago de Castro Silva
  2. Anna Bianchi
  3. Nilesh U Deshpande
  4. Prateek Sharma
  5. Siddharth Mehra
  6. Vanessa Tonin Garrido
  7. Shannon Jacqueline Saigh
  8. Jonathan England
  9. Peter Joel Hosein
  10. Deukwoo Kwon
  11. Nipun B Merchant
  12. Jashodeep Datta  Is a corresponding author
  1. University of Miami, United States
  2. University of Nebraska Medical Center, United States
  3. Sylvester Comprehensive Cancer Center, United States
  4. The University of Texas Health Science Center at Houston, United States

Abstract

Background: Partial/complete pathologic response following neoadjuvant chemotherapy (NAC) in pancreatic cancer (PDAC) patients undergoing pancreatectomy is associated with improved survival. We sought to determine whether neutrophil-to-lymphocyte ratio (NLR) dynamics predict pathologic response following chemotherapy in PDAC, and if manipulating NLR impacts chemosensitivity in preclinical models and uncovers potential mechanistic underpinnings underlying these effects.

Methods: Pathologic response in PDAC patients (n=94) undergoing NAC and pancreatectomy (7/2015-12/2019) was dichotomized as partial/complete or poor/absent. Bootstrap-validated multivariable models assessed associations between pre-chemotherapy NLR (%neutrophils÷%lymphocytes) or NLR dynamics during chemotherapy (ΔNLR=pre-surgery-pre-chemotherapy NLR) and pathologic response, disease-free survival (DFS), and overall survival (OS). To preclinically model effects of NLR attenuation on chemosensitivity, Ptf1aCre/+; KrasLSL-G12D/+;Tgfbr2flox/flox (PKT) mice and C57BL/6 mice orthotopically injected with KrasLSL-G12D/+;Trp53LSL-R172H/+;Pdx1Cre (KPC) cells were randomized to vehicle, gemcitabine/paclitaxel alone, and NLR-attenuating anti-Ly6G with/without gemcitabine/paclitaxel treatment.

Results: In 94 PDAC patients undergoing NAC (median:4 months), pre-chemotherapy NLR (P<0.001) and ΔNLR attenuation during NAC (P=0.002) were independently associated with partial/complete pathologic response. An NLR score=pre-chemotherapy NLR+ΔNLR correlated with DFS (P=0.006) and OS (P=0.002). Upon preclinical modeling, combining NLR-attenuating anti-Ly6G treatment with gemcitabine/paclitaxel-compared with gemcitabine/paclitaxel or anti-Ly6G alone-not only significantly reduced tumor burden and metastatic outgrowth, but also augmented tumor-infiltrating CD107a+-degranulating CD8+ T-cells (P<0.01) while dampening inflammatory cancer-associated fibroblast (CAF) polarization (P=0.006) and chemoresistant IL-6/STAT-3 signaling in vivo. Neutrophil-derived IL-1β emerged as a novel mediator of stromal inflammation, inducing inflammatory CAF polarization and CAF-tumor cell IL-6/STAT-3 signaling in ex vivo co-cultures.

Conclusions: Therapeutic strategies to mitigate neutrophil-CAF-tumor cell IL-1β/IL-6/STAT-3 signaling during NAC may improve pathologic responses and/or survival in PDAC.

Funding: Supported by KL2 career development grant by Miami CTSI under NIH Award UL1TR002736, Stanley Glaser Foundation, American College of Surgeons Franklin Martin Career Development Award, and Association for Academic Surgery Joel J. Roslyn Faculty Award (to J. Datta); NIH R01 CA161976 (to N.B. Merchant); and NCI/NIH Award P30CA240139 (to J. Datta and N.B. Merchant).

Data availability

Clinicodemographic data utilized in this analysis can be tracked back to individual patients (e.g., age, CA19-9 values, pathologic response score) despite deidentification. Since these comprise protected health information from human subjects, a limited deidentified dataset with only relevant data to allow reproduction of major findings are provided. All relevant source data from in vitro experiments have also been provided.

Article and author information

Author details

  1. Iago de Castro Silva

    Department of Surgery, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Anna Bianchi

    Department of Surgery, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Nilesh U Deshpande

    Department of Surgery, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Prateek Sharma

    Department of Surgery, University of Nebraska Medical Center, Omaha, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Siddharth Mehra

    Department of Surgery, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Vanessa Tonin Garrido

    Department of Surgery, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Shannon Jacqueline Saigh

    Sylvester Comprehensive Cancer Center, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Jonathan England

    Department of Pathology, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Peter Joel Hosein

    Department of Medicine, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Deukwoo Kwon

    Department of Public Health Sciences, The University of Texas Health Science Center at Houston, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Nipun B Merchant

    Department of Surgery, University of Miami, Miami, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Jashodeep Datta

    Department of Surgery, University of Miami, Miami, United States
    For correspondence
    jash.datta@med.miami.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2869-1571

Funding

National Institutes of Health (UL1TR002736)

  • Jashodeep Datta

American College of Surgeons (Franklin H. Martin Research Fellowship)

  • Jashodeep Datta

Association for Academic Surgery Foundation (Joel J. Roslyn Faculty Award)

  • Jashodeep Datta

University of Miami (Stanley Glaser Foundation Award)

  • Jashodeep Datta

National Cancer Institute (P30CA240139)

  • Nipun B Merchant
  • Jashodeep Datta

National Cancer Institute (R01CA161976)

  • Nipun B Merchant

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: This study was strictly performed in agreement with all the recommendations stablished in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All animal work was performed following the approved Institutional Animal Care and Use Committee (IACUC) protocol (#21-057) of the University of Miami, and supervised by the Division of Veterinary Resources (DVR). All surgical procedures were performed under general anesthesia, analgesic drugs were administered postoperatively, and every effort was made to minimize any form of animal suffering.

Human subjects: -Informed consent was not necessary since tissue sections were accrued previously under a cancer center-wide biospecimen protocol-Institution Review Board /PRMC/site disease group approval for this study was obtained under protocol 20200123

Copyright

© 2022, de Castro Silva et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Iago de Castro Silva
  2. Anna Bianchi
  3. Nilesh U Deshpande
  4. Prateek Sharma
  5. Siddharth Mehra
  6. Vanessa Tonin Garrido
  7. Shannon Jacqueline Saigh
  8. Jonathan England
  9. Peter Joel Hosein
  10. Deukwoo Kwon
  11. Nipun B Merchant
  12. Jashodeep Datta
(2022)
Neutrophil-mediated fibroblast-tumor cell IL-6/STAT-3 signaling underlies the association between neutrophil-to-lymphocyte ratio dynamics and chemotherapy response in localized pancreatic cancer: a hybrid clinical-preclinical study
eLife 11:e78921.
https://doi.org/10.7554/eLife.78921

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https://doi.org/10.7554/eLife.78921

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