1. Immunology and Inflammation
  2. Medicine

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
https://doi.org/10.7554/eLife.78921
Share this article
Cite this article
  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
  2. Download BibTeX
  3. Download .RIS

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.

Metrics

  • 1,655
    views
  • 378
    downloads
  • 23
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  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

Share this article

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

Categories and tags

Research organisms

Further reading

    1. Immunology and Inflammation

    Follicular helper- and peripheral helper-like T cells drive autoimmune disease in human immune system mice

    Mohsen Khosravi-Maharlooei, Andrea Vecchione ... Megan Sykes
    Research Article

    Human immune system (HIS) mice constructed in various ways are widely used for investigations of human immune responses to pathogens, transplants, and immunotherapies. In HIS mice that generate T cells de novo from hematopoietic progenitors, T cell-dependent multisystem autoimmune disease occurs, most rapidly when the human T cells develop in the native NOD.Cg- Prkdcscid Il2rgtm1Wjl (NSG) mouse thymus, where negative selection is abnormal. Disease develops very late when human T cells develop in human fetal thymus grafts, where robust negative selection is observed. We demonstrate here that PD-1+CD4+ peripheral (Tph) helper-like and follicular (Tfh) helper-like T cells developing in HIS mice can induce autoimmune disease. Tfh-like cells were more prominent in HIS mice with a mouse thymus, in which the highest levels of IgG were detected in plasma, compared to those with a human thymus. While circulating IgG and IgM antibodies were autoreactive to multiple mouse antigens, in vivo depletion of B cells and antibodies did not delay the development of autoimmune disease. Conversely, adoptive transfer of enriched Tfh- or Tph-like cells induced disease and autoimmunity-associated B cell phenotypes in recipient mice containing autologous human APCs without T cells. Tfh/Tph cells from mice with a human thymus expanded and induced disease more rapidly than those originating in a murine thymus, implicating HLA-restricted T cell-APC interactions in this process. Since Tfh, Tph, autoantibodies, and lymphopenia-induced proliferation (LIP) have all been implicated in various forms of human autoimmune disease, the observations here provide a platform for the further dissection of human autoimmune disease mechanisms and therapies.

    1. Immunology and Inflammation

    A host enzyme reduces metabolic dysfunction-associated steatotic liver disease (MASLD) by inactivating intestinal lipopolysaccharide

    Zhiyan Wang, Nore Ojogun ... Mingfang Lu
    Research Article

    The incidence of metabolic dysfunction-associated steatotic liver disease (MASLD) has been increasing worldwide. Since gut-derived bacterial lipopolysaccharides (LPS) can travel via the portal vein to the liver and play an important role in producing hepatic pathology, it seemed possible that (1) LPS stimulates hepatic cells to accumulate lipid, and (2) inactivating LPS can be preventive. Acyloxyacyl hydrolase (AOAH), the eukaryotic lipase that inactivates LPS and oxidized phospholipids, is produced in the intestine, liver, and other organs. We fed mice either normal chow or a high-fat diet for 28 weeks and found that Aoah-/- mice accumulated more hepatic lipid than did Aoah+/+ mice. In young mice, before increased hepatic fat accumulation was observed, Aoah-/- mouse livers increased their abundance of sterol regulatory element-binding protein 1, and the expression of its target genes that promote fatty acid synthesis. Aoah-/- mice also increased hepatic expression of Cd36 and Fabp3, which mediate fatty acid uptake, and decreased expression of fatty acid-oxidation-related genes Acot2 and Ppara. Our results provide evidence that increasing AOAH abundance in the gut, bloodstream, and/or liver may be an effective strategy for preventing or treating MASLD.

    1. Immunology and Inflammation
    2. Microbiology and Infectious Disease

    Monoclonal antibodies derived from B cells in subjects with cystic fibrosis reduce Pseudomonas aeruginosa burden in mice

    Malika Hale, Kennidy K Takehara ... Marion Pepper
    Research Article

    Pseudomonas aeruginosa (PA) is an opportunistic, frequently multidrug-resistant pathogen that can cause severe infections in hospitalized patients. Antibodies against the PA virulence factor, PcrV, protect from death and disease in a variety of animal models. However, clinical trials of PcrV-binding antibody-based products have thus far failed to demonstrate benefit. Prior candidates were derivations of antibodies identified using protein-immunized animal systems and required extensive engineering to optimize binding and/or reduce immunogenicity. Of note, PA infections are common in people with cystic fibrosis (pwCF), who are generally believed to mount normal adaptive immune responses. Here, we utilized a tetramer reagent to detect and isolate PcrV-specific B cells in pwCF and, via single-cell sorting and paired-chain sequencing, identified the B cell receptor (BCR) variable region sequences that confer PcrV-specificity. We derived multiple high affinity anti-PcrV monoclonal antibodies (mAbs) from PcrV-specific B cells across three donors, including mAbs that exhibit potent anti-PA activity in a murine pneumonia model. This robust strategy for mAb discovery expands what is known about PA-specific B cells in pwCF and yields novel mAbs with potential for future clinical use.