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
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Curated by eLife
Evaluation Summary:
Iago De Castro et al, constitute an exciting study conveying to readers that neutrophil-to-lymphocyte ratio dynamics predict pancreatic cancer pathologic response to neoadjuvant therapy. Specifically, the authors aim to determine the effect of gemcitabine/paclitaxel and anti-Ly6G treatment on stromal T cells and CAF populations. They conclude that neutrophil-to-lymphocyte ratios are associated with survival following this treatment and use animal models to show metastatic effects allied to it. The authors attempt to convey that microenvironmental neutrophils could play a causal role in pancreatic cancer chemoresistance. It was agreed that while the discoveries are very interesting, the public could benefit from the authors improving: the relevance to the human condition, providing a stronger link to fibroblastic cell functional transitions, broadening the discussion regarding previous/related published studies, and strengthening the anti-Ly6G specificity proof within the provided data.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #2 agreed to share their name with the authors.)
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Abstract
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, Ptf1a Cre/+ ; Kras LSL-G12D/+ ;Tgfbr2 flox/flox (PKT) mice and C57BL/6 mice orthotopically injected with Kras LSL-G12D/+ ;Trp53 LSL-R172H/+ ;Pdx1 Cre (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).
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Evaluation Summary:
Iago De Castro et al, constitute an exciting study conveying to readers that neutrophil-to-lymphocyte ratio dynamics predict pancreatic cancer pathologic response to neoadjuvant therapy. Specifically, the authors aim to determine the effect of gemcitabine/paclitaxel and anti-Ly6G treatment on stromal T cells and CAF populations. They conclude that neutrophil-to-lymphocyte ratios are associated with survival following this treatment and use animal models to show metastatic effects allied to it. The authors attempt to convey that microenvironmental neutrophils could play a causal role in pancreatic cancer chemoresistance. It was agreed that while the discoveries are very interesting, the public could benefit from the authors improving: the relevance to the human condition, providing a stronger link to fibroblastic cell …
Evaluation Summary:
Iago De Castro et al, constitute an exciting study conveying to readers that neutrophil-to-lymphocyte ratio dynamics predict pancreatic cancer pathologic response to neoadjuvant therapy. Specifically, the authors aim to determine the effect of gemcitabine/paclitaxel and anti-Ly6G treatment on stromal T cells and CAF populations. They conclude that neutrophil-to-lymphocyte ratios are associated with survival following this treatment and use animal models to show metastatic effects allied to it. The authors attempt to convey that microenvironmental neutrophils could play a causal role in pancreatic cancer chemoresistance. It was agreed that while the discoveries are very interesting, the public could benefit from the authors improving: the relevance to the human condition, providing a stronger link to fibroblastic cell functional transitions, broadening the discussion regarding previous/related published studies, and strengthening the anti-Ly6G specificity proof within the provided data.
(This preprint has been reviewed by eLife. We include the public reviews from the reviewers here; the authors also receive private feedback with suggested changes to the manuscript. Reviewer #2 agreed to share their name with the authors.)
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Reviewer #1 (Public Review):
This is an interesting study, addressing a timely question of the crosstalk between cancer, immune, and stromal cell populations in the tumor microenvironment, and the effect of therapy on the tumor microenvironment. The authors were aiming to show that the ratio between neutrophils and lymphocytes could predict treatment responses in pancreatic cancer. They indeed show that there is an association between the Neutrophil to lymphocyte ratio (NLR) and treatment outcome, suggesting that this could be a predictive marker. They go on to use a mouse model to perturb the NLR and combine this with treatment similar to that used in the clinic and find that targeting neutrophils affects tumor growth, suggesting a costive and not the only correlative role. Finally, they show that this could be mediated through the …
Reviewer #1 (Public Review):
This is an interesting study, addressing a timely question of the crosstalk between cancer, immune, and stromal cell populations in the tumor microenvironment, and the effect of therapy on the tumor microenvironment. The authors were aiming to show that the ratio between neutrophils and lymphocytes could predict treatment responses in pancreatic cancer. They indeed show that there is an association between the Neutrophil to lymphocyte ratio (NLR) and treatment outcome, suggesting that this could be a predictive marker. They go on to use a mouse model to perturb the NLR and combine this with treatment similar to that used in the clinic and find that targeting neutrophils affects tumor growth, suggesting a costive and not the only correlative role. Finally, they show that this could be mediated through the stromal compartment since this treatment affects the ratio of inflammatory to myofibroblastic CAFs.
The main strength of the paper is in tying together neutrophils, lymphocytes, and CAFs and showing how these populations affect each other. The correlations in human patients are promising and the regulation of CAF transitions is interesting.
While the correlation between NLR and survival is convincing and strong, the relevance of CAF transitions to this effect in human patients is weak, and shown only in mice and not in humans. Also in the mouse, the evidence for CAF transitions should be strengthened to support the authors' full conclusions.
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Reviewer #2 (Public Review):
The overall goal of this study by de Castro Silva and colleagues is certainly significant and clinically relevant in aiming to identify a biomarker to predict responses to neoadjuvant chemotherapy among PDAC patients. The authors' conclusions, that pre-treatment NLR, as well as change in NLR over the course of neoadjuvant chemotherapy, associate with partial/complete pathologic responses, are supported by the data and likely to be of broad interest to the pancreatic cancer research community. Additional strengths include the novel and compelling implication of Ly6G+ cells in chemoresistance in a preclinical model of PDAC as well as the roles of these cells in the regulation of CAF phenotypes and immune-modulatory gene expression in a paracrine manner, also supported by convincing results in the manuscript. …
Reviewer #2 (Public Review):
The overall goal of this study by de Castro Silva and colleagues is certainly significant and clinically relevant in aiming to identify a biomarker to predict responses to neoadjuvant chemotherapy among PDAC patients. The authors' conclusions, that pre-treatment NLR, as well as change in NLR over the course of neoadjuvant chemotherapy, associate with partial/complete pathologic responses, are supported by the data and likely to be of broad interest to the pancreatic cancer research community. Additional strengths include the novel and compelling implication of Ly6G+ cells in chemoresistance in a preclinical model of PDAC as well as the roles of these cells in the regulation of CAF phenotypes and immune-modulatory gene expression in a paracrine manner, also supported by convincing results in the manuscript. Weaknesses were also noted, including a lack of discussion of several relevant prior studies of NLR in PDAC in the neoadjuvant setting, lack of specificity of the Ly6G neutralizing antibody for neutrophils, and use of a single mouse model for preclinical evaluation. Addressing these weaknesses would strengthen this study and increase its impact on the field.
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