Oncogenic Ras and ΔNp63α cooperate to recruit immunosuppressive polymorphonuclear myeloid-derived suppressor cells in a mouse model of squamous cancer pathogenesis

Sakakibara, Nozomi; Clavijo, Paúl E.; Sievers, Cem; Gray, Veronica C.; King, Kathryn E.; George, Andrea L.; Ponnamperuma, Roshini M.; Walter, Beatriz A.; Chen, Zhong; Van Waes, Carter; Allen, Clint T.; Weinberg, Wendy C.

doi:10.3389/fimmu.2023.1200970

Oncogenic Ras and ΔNp63α cooperate to recruit immunosuppressive polymorphonuclear myeloid-derived suppressor cells in a mouse model of squamous cancer pathogenesis

Journal Article · Thu Aug 10 00:00:00 EDT 2023 · Frontiers in Immunology

DOI:https://doi.org/10.3389/fimmu.2023.1200970· OSTI ID:1994573

Sakakibara, Nozomi; Clavijo, Paúl E.; Sievers, Cem; Gray, Veronica C.; King, Kathryn E.; George, Andrea L.; Ponnamperuma, Roshini M.; Walter, Beatriz A.; Chen, Zhong; Van Waes, Carter; Allen, Clint T.; Weinberg, Wendy C.

Introduction

Amplification of human chromosome 3q26-29, which encodes oncoprotein ΔNp63 among other isoforms of the p63 family, is a feature common to squamous cell carcinomas (SCCs) of multiple tissue origins. Along with overexpression of ΔNp63, activation of the protooncogene, RAS , whether by overexpression or oncogenic mutation, is frequently observed in many cancers. In this study, analysis of transcriptome data from The Cancer Genome Atlas (TCGA) demonstrated that expression of TP63 mRNA , particularly ΔNp63 isoforms, and HRAS are significantly elevated in advanced squamous cell carcinomas of the head and neck (HNSCCs), suggesting pathological significance. However, how co-overexpressed ΔNp63 and HRAS affect the immunosuppressive tumor microenvironment (TME) is incompletely understood.

Methods

Here, we established and characterized an immune competent mouse model using primary keratinocytes with retroviral-mediated overexpression of ΔNp63α and constitutively activated HRAS (v-ras ^Ha G12R) to evaluate the role of these oncogenes in the immune TME.

Results

In this model, orthotopic grafting of wildtype syngeneic keratinocytes expressing both v-ras ^Ha and elevated levels of ΔNp63α consistently yield carcinomas in syngeneic hosts, while cells expressing v-ras ^Ha alone yield predominantly papillomas. We found that polymorphonuclear (PMN) myeloid cells, experimentally validated to be immunosuppressive and thus representing myeloid-derived suppressor cells (PMN-MDSCs), were significantly recruited into the TME of carcinomas arising early following orthotopic grafting of ΔNp63α/v-ras ^Ha -expressing keratinocytes. ΔNp63α/v-ras ^Ha -driven carcinomas expressed higher levels of chemokines implicated in recruitment of MDSCs compared to v-ras ^Ha -initiated tumors, providing a heretofore undescribed link between ΔNp63α/HRAS-driven carcinomas and the development of an immunosuppressive TME.

Conclusion

These results support the utilization of a genetic carcinogenesis model harboring specific genomic drivers of malignancy to study mechanisms underlying the development of local immunosuppression.

View Journal Article

Sponsoring Organization:: USDOE

OSTI ID:: 1994573

Alternate ID(s):: OSTI ID: 2425753

Journal Information:: Frontiers in Immunology, Journal Name: Frontiers in Immunology Vol. 14; ISSN 1664-3224

Publisher:: Frontiers Media SACopyright Statement

Country of Publication:: Switzerland

Language:: English

References (61)

Interpretation of cellular proliferation data: Avoid the panglossian Roederer, Mario Cytometry Part A, Vol. 79A, Issue 2 https://doi.org/10.1002/cyto.a.21010	journal	January 2011
Intersection of the p63 and NF‐κB pathways in epithelial homeostasis and disease King, Kathryn E.; George, Andrea L.; Sakakibara, Nozomi Molecular Carcinogenesis, Vol. 58, Issue 9 https://doi.org/10.1002/mc.23081	journal	July 2019
The transcriptional regulatory function of p53 is essential for suppression of mouse skin carcinogenesis and can be dissociated from effects on TGF-β-mediated growth regulation Ponnamperuma, Roshini M.; King, Kathryn E.; Elsir, Tamador The Journal of Pathology, Vol. 219, Issue 2 https://doi.org/10.1002/path.2600	journal	July 2009
Epidermal overexpression of transgenic ΔNp63 promotes type 2 immune and myeloid inflammatory responses and hyperplasia via NF-κB activation: Δp63 promotes skin inflammation via NF-κB activation Du, Jihui; Romano, Rose-Anne; Si, Han The Journal of Pathology, Vol. 232, Issue 3 https://doi.org/10.1002/path.4302	journal	January 2014
CD4 and CD8 T lymphocyte interplay in controlling tumor growth Ostroumov, Dmitrij; Fekete-Drimusz, Nora; Saborowski, Michael Cellular and Molecular Life Sciences, Vol. 75, Issue 4 https://doi.org/10.1007/s00018-017-2686-7	journal	October 2017
Master regulatory role of p63 in epidermal development and disease Soares, Eduardo; Zhou, Huiqing Cellular and Molecular Life Sciences, Vol. 75, Issue 7 https://doi.org/10.1007/s00018-017-2701-z	journal	November 2017
Characterization of specific p63 and p63-N-terminal isoform antibodies and their application for immunohistochemistry Nekulova, Marta; Holcakova, Jitka; Nenutil, Rudolf Virchows Archiv, Vol. 463, Issue 3 https://doi.org/10.1007/s00428-013-1459-4	journal	July 2013
Multiplatform Analysis of 12 Cancer Types Reveals Molecular Classification within and across Tissues of Origin Hoadley, Katherine A.; Yau, Christina; Wolf, Denise M. Cell, Vol. 158, Issue 4 https://doi.org/10.1016/j.cell.2014.06.049	journal	August 2014
Single-Cell Transcriptomic Analysis of Primary and Metastatic Tumor Ecosystems in Head and Neck Cancer Puram, Sidharth V.; Tirosh, Itay; Parikh, Anuraag S. Cell, Vol. 171, Issue 7 https://doi.org/10.1016/j.cell.2017.10.044	journal	December 2017
Cell-of-Origin Patterns Dominate the Molecular Classification of 10,000 Tumors from 33 Types of Cancer Hoadley, Katherine A.; Yau, Christina; Hinoue, Toshinori Cell, Vol. 173, Issue 2 https://doi.org/10.1016/j.cell.2018.03.022	journal	April 2018
NF-κB Functions in Tumor Initiation by Suppressing the Surveillance of Both Innate and Adaptive Immune Cells Wang, David J.; Ratnam, Nivedita M.; Byrd, John C. Cell Reports, Vol. 9, Issue 1 https://doi.org/10.1016/j.celrep.2014.08.049	journal	October 2014
Genomic, Pathway Network, and Immunologic Features Distinguishing Squamous Carcinomas Campbell, Joshua D.; Yau, Christina; Bowlby, Reanne Cell Reports, Vol. 23, Issue 1 https://doi.org/10.1016/j.celrep.2018.03.063	journal	April 2018
The Nature of Myeloid-Derived Suppressor Cells in the Tumor Microenvironment Kumar, Vinit; Patel, Sima; Tcyganov, Evgenii Trends in Immunology, Vol. 37, Issue 3 https://doi.org/10.1016/j.it.2016.01.004	journal	March 2016
Elevated ΔNp63α Levels Facilitate Epidermal and Biliary Oncogenic Transformation Devos, Michael; Gilbert, Barbara; Denecker, Geertrui Journal of Investigative Dermatology, Vol. 137, Issue 2 https://doi.org/10.1016/j.jid.2016.09.026	journal	February 2017
Co-Activation of Glucocorticoid Receptor and Peroxisome Proliferator–Activated Receptor-γ in Murine Skin Prevents Worsening of Atopic March Deckers, Julie; Bougarne, Nadia; Mylka, Viacheslav Journal of Investigative Dermatology, Vol. 138, Issue 6 https://doi.org/10.1016/j.jid.2017.12.023	journal	June 2018
Overcoming barriers to effective immunotherapy: MDSCs, TAMs, and Tregs as mediators of the immunosuppressive microenvironment in head and neck cancer Davis, Ruth J.; Van Waes, Carter; Allen, Clint T. Oral Oncology, Vol. 58 https://doi.org/10.1016/j.oraloncology.2016.05.002	journal	July 2016
ΔNp63α Is an Oncogene that Targets Chromatin Remodeler Lsh to Drive Skin Stem Cell Proliferation and Tumorigenesis Keyes, William M.; Pecoraro, Matteo; Aranda, Victoria Cell Stem Cell, Vol. 8, Issue 2 https://doi.org/10.1016/j.stem.2010.12.009	journal	February 2011
An activated Harvey ras oncogene produces benign tumours on mouse epidermal tissue Roop, Dennis R.; Lowy, Douglas R.; Tambourin, Pierre E. Nature, Vol. 323, Issue 6091 https://doi.org/10.1038/323822a0	journal	October 1986
p63, a Story of Mice and Men Vanbokhoven, Hans; Melino, Gerry; Candi, Eleonora Journal of Investigative Dermatology, Vol. 131, Issue 6 https://doi.org/10.1038/jid.2011.84	journal	June 2011
Comprehensive genomic characterization of head and neck squamous cell carcinomas No authors listed Nature, Vol. 517, Issue 7536, p. 576-582 https://doi.org/10.1038/nature14129	journal	January 2015
Cross-species identification of genomic drivers of squamous cell carcinoma development across preneoplastic intermediates Chitsazzadeh, Vida; Coarfa, Cristian; Drummond, Jennifer A. Nature Communications, Vol. 7, Issue 1 https://doi.org/10.1038/ncomms12601	journal	August 2016
Isolation and short-term culture of primary keratinocytes, hair follicle populations and dermal cells from newborn mice and keratinocytes from adult mice for in vitro analysis and for grafting to immunodeficient mice Lichti, Ulrike; Anders, Joanna; Yuspa, Stuart H. Nature Protocols, Vol. 3, Issue 5 https://doi.org/10.1038/nprot.2008.50	journal	April 2008
Chemokines in the cancer microenvironment and their relevance in cancer immunotherapy Nagarsheth, Nisha; Wicha, Max S.; Zou, Weiping Nature Reviews Immunology, Vol. 17, Issue 9 https://doi.org/10.1038/nri.2017.49	journal	May 2017
TNF-α modulates genome-wide redistribution of ΔNp63α/TAp73 and NF-κB cREL interactive binding on TP53 and AP-1 motifs to promote an oncogenic gene program in squamous cancer Si, H.; Lu, H.; Yang, X. Oncogene, Vol. 35, Issue 44 https://doi.org/10.1038/onc.2016.112	journal	May 2016
The genomic landscape of cutaneous SCC reveals drivers and a novel azathioprine associated mutational signature Inman, Gareth J.; Wang, Jun; Nagano, Ai Nature Communications, Vol. 9, Issue 1 https://doi.org/10.1038/s41467-018-06027-1	journal	September 2018
Myeloid-derived suppressor cells coming of age Veglia, Filippo; Perego, Michela; Gabrilovich, Dmitry Nature Immunology, Vol. 19, Issue 2 https://doi.org/10.1038/s41590-017-0022-x	journal	January 2018
Growth Regulated Oncogene-α expression by murine squamous cell carcinoma promotes tumor growth, metastasis, leukocyte infiltration and angiogenesis by a host CXC Receptor-2 dependent mechanism Loukinova, Elena; Dong, Gang; Enamorado-Ayalya, Ileana Oncogene, Vol. 19, Issue 31 https://doi.org/10.1038/sj.onc.1203687	journal	July 2000
p63α and ΔNp63α can induce cell cycle arrest and apoptosis and differentially regulate p53 target genes Dohn, Michael; Zhang, Shungzhen; Chen, Xinbin Oncogene, Vol. 20, Issue 25 https://doi.org/10.1038/sj.onc.1204427	journal	May 2001
ΔNp63α functions as both a positive and a negative transcriptional regulator and blocks in vitro differentiation of murine keratinocytes King, Kathryn E.; Ponnamperuma, Roshini M.; Yamashita, Toshiharu Oncogene, Vol. 22, Issue 23 https://doi.org/10.1038/sj.onc.1206536	journal	June 2003
Oncogenic Ha-Ras-induced Signaling Activates NF-κB Transcriptional Activity, Which Is Required for Cellular Transformation Finco, Timothy S.; Westwick, John K.; Norris, Jacqueline L. Journal of Biological Chemistry, Vol. 272, Issue 39 https://doi.org/10.1074/jbc.272.39.24113	journal	September 1997
Inhibition of tumor growth by elimination of granulocytes. Pekarek, L. A.; Starr, B. A.; Toledano, A. Y. The Journal of experimental medicine, Vol. 181, Issue 1 https://doi.org/10.1084/jem.181.1.435	journal	January 1995
IL-1R–MyD88 signaling in keratinocyte transformation and carcinogenesis Cataisson, Christophe; Salcedo, Rosalba; Hakim, Shakeeb Journal of Experimental Medicine, Vol. 209, Issue 9 https://doi.org/10.1084/jem.20101355	journal	August 2012
Senescence delay and repression of p16 INK4a by Lsh via recruitment of histone deacetylases in human diploid fibroblasts Zhou, Rui; Han, Limin; Li, Guodong Nucleic Acids Research, Vol. 37, Issue 15 https://doi.org/10.1093/nar/gkp533	journal	June 2009
canSAR: an integrated cancer public translational research and drug discovery resource Halling-Brown, Mark D.; Bulusu, Krishna C.; Patel, Mishal Nucleic Acids Research, Vol. 40, Issue D1 https://doi.org/10.1093/nar/gkr881	journal	October 2011
canSAR: an updated cancer research and drug discovery knowledgebase Tym, Joseph E.; Mitsopoulos, Costas; Coker, Elizabeth A. Nucleic Acids Research, Vol. 44, Issue D1 https://doi.org/10.1093/nar/gkv1030	journal	December 2015
canSAR: update to the cancer translational research and drug discovery knowledgebase Coker, Elizabeth A.; Mitsopoulos, Costas; Tym, Joesph E. Nucleic Acids Research, Vol. 47, Issue D1 https://doi.org/10.1093/nar/gky1129	journal	November 2018
Loss of transforming growth factor-β type II receptor promotes metastatic head-and-neck squamous cell carcinoma Lu, Shi-Long; Herrington, Heather; Reh, Douglas Genes & Development, Vol. 20, Issue 10 https://doi.org/10.1101/gad.1413306	journal	May 2006
Integrative Analysis of Complex Cancer Genomics and Clinical Profiles Using the cBioPortal Gao, J.; Aksoy, B. A.; Dogrusoz, U. Science Signaling, Vol. 6, Issue 269 https://doi.org/10.1126/scisignal.2004088	journal	March 2013
MET signaling in keratinocytes activates EGFR and initiates squamous carcinogenesis Cataisson, Christophe; Michalowski, Aleksandra M.; Shibuya, Kelly Science Signaling, Vol. 9, Issue 433 https://doi.org/10.1126/scisignal.aaf5106	journal	June 2016
The p53 Homologue Np63 Interacts with the Nuclear Factor- B Pathway to Modulate Epithelial Cell Growth King, K. E.; Ponnamperuma, R. M.; Allen, C. Cancer Research, Vol. 68, Issue 13 https://doi.org/10.1158/0008-5472.CAN-07-6123	journal	July 2008
Inducible Cutaneous Inflammation Reveals a Protumorigenic Role for Keratinocyte CXCR2 in Skin Carcinogenesis Cataisson, Christophe; Ohman, Rebecca; Patel, Gopal Cancer Research, Vol. 69, Issue 1 https://doi.org/10.1158/0008-5472.CAN-08-2490	journal	December 2008
Np63 Versatilely Regulates a Broad NF- B Gene Program and Promotes Squamous Epithelial Proliferation, Migration, and Inflammation Yang, X.; Lu, H.; Yan, B. Cancer Research, Vol. 71, Issue 10 https://doi.org/10.1158/0008-5472.CAN-10-3445	journal	May 2011
TNF- Promotes c-REL/ Np63 Interaction and TAp73 Dissociation from Key Genes That Mediate Growth Arrest and Apoptosis in Head and Neck Cancer Lu, H.; Yang, X.; Duggal, P. Cancer Research, Vol. 71, Issue 21 https://doi.org/10.1158/0008-5472.CAN-11-2460	journal	September 2011
Osteopontin Shapes Immunosuppression in the Metastatic Niche Sangaletti, Sabina; Tripodo, Claudio; Sandri, Sara Cancer Research, Vol. 74, Issue 17 https://doi.org/10.1158/0008-5472.CAN-13-3334	journal	September 2014
CD38-Expressing Myeloid-Derived Suppressor Cells Promote Tumor Growth in a Murine Model of Esophageal Cancer Karakasheva, Tatiana A.; Waldron, Todd J.; Eruslanov, Evgeniy Cancer Research, Vol. 75, Issue 19 https://doi.org/10.1158/0008-5472.CAN-14-3639	journal	September 2015
Anti-PD-L1 Efficacy Can Be Enhanced by Inhibition of Myeloid-Derived Suppressor Cells with a Selective Inhibitor of PI3Kδ/γ Davis, Ruth J.; Moore, Ellen C.; Clavijo, Paul E. Cancer Research, Vol. 77, Issue 10 https://doi.org/10.1158/0008-5472.CAN-16-2534	journal	May 2017
Mutational Landscape of Aggressive Cutaneous Squamous Cell Carcinoma Pickering, Curtis R.; Zhou, Jane H.; Lee, J. Jack Clinical Cancer Research, Vol. 20, Issue 24 https://doi.org/10.1158/1078-0432.CCR-14-1768	journal	December 2014
Tipifarnib as a Precision Therapy for HRAS-Mutant Head and Neck Squamous Cell Carcinomas Gilardi, Mara; Wang, Zhiyong; Proietto, Marco Molecular Cancer Therapeutics, Vol. 19, Issue 9 https://doi.org/10.1158/1535-7163.MCT-19-0958	journal	September 2020
The cBio Cancer Genomics Portal: An Open Platform for Exploring Multidimensional Cancer Genomics Data: Figure 1. Cerami, Ethan; Gao, Jianjiong; Dogrusoz, Ugur Cancer Discovery, Vol. 2, Issue 5 https://doi.org/10.1158/2159-8290.CD-12-0095	journal	May 2012
Host Immunity Following Near-Infrared Photoimmunotherapy Is Enhanced with PD-1 Checkpoint Blockade to Eradicate Established Antigenic Tumors Nagaya, Tadanobu; Friedman, Jay; Maruoka, Yasuhiro Cancer Immunology Research, Vol. 7, Issue 3 https://doi.org/10.1158/2326-6066.CIR-18-0546	journal	March 2019
Murine models of HRAS-mediated cutaneous skeletal hypophosphatemia syndrome suggest bone as the FGF23 excess source Ovejero, Diana; Michel, Zachary; Cataisson, Christophe Journal of Clinical Investigation, Vol. 133, Issue 9 https://doi.org/10.1172/JCI159330	journal	May 2023
FGFR2 signaling underlies p63 oncogenic function in squamous cell carcinoma Ramsey, Matthew R.; Wilson, Catherine; Ory, Benjamin Journal of Clinical Investigation, Vol. 123, Issue 8 https://doi.org/10.1172/JCI68899	journal	July 2013
ΔNp63-driven recruitment of myeloid-derived suppressor cells promotes metastasis in triple-negative breast cancer Kumar, Sushil; Wilkes, David W.; Samuel, Nina Journal of Clinical Investigation, Vol. 128, Issue 11 https://doi.org/10.1172/JCI99673	journal	September 2018
Abnormal hair follicle development and altered cell fate of follicular keratinocytes in transgenic mice expressing ΔNp63α Romano, Rose-Anne; Smalley, Kirsten; Liu, Song Development, Vol. 137, Issue 9 https://doi.org/10.1242/dev.045427	journal	May 2010
Dysregulated ΔNp63α Inhibits Expression of Ink4a/arf, Blocks Senescence, and Promotes Malignant Conversion of Keratinocytes Ha, Linan; Ponnamperuma, Roshini M.; Jay, Steven PLoS ONE, Vol. 6, Issue 7 https://doi.org/10.1371/journal.pone.0021877	journal	July 2011
Epidermal p65/ NF ‐κB signalling is essential for skin carcinogenesis Kim, Chun; Pasparakis, Manolis EMBO Molecular Medicine, Vol. 6, Issue 7 https://doi.org/10.15252/emmm.201303541	journal	June 2014
Induction of ΔNp63 by the Newly Identified Keratinocyte-Specific Transforming Growth Factor β Signaling Pathway with Smad2 and IκB Kinase α in Squamous Cell Carcinoma Fukunishi, Nahoko; Katoh, Iyoko; Tomimori, Yoshiya Neoplasia, Vol. 12, Issue 12 https://doi.org/10.1593/neo.101054	journal	December 2010
Targeting Myeloid-Derived Suppressor Cells to Bypass Tumor-Induced Immunosuppression Fleming, Viktor; Hu, Xiaoying; Weber, Rebekka Frontiers in Immunology, Vol. 9 https://doi.org/10.3389/fimmu.2018.00398	journal	March 2018
Chemokines Modulate Immune Surveillance in Tumorigenesis, Metastasis, and Response to Immunotherapy Vilgelm, Anna E.; Richmond, Ann Frontiers in Immunology, Vol. 10 https://doi.org/10.3389/fimmu.2019.00333	journal	February 2019
Molecular Mechanisms of p63-Mediated Squamous Cancer Pathogenesis Moses, Michael A.; George, Andrea L.; Sakakibara, Nozomi International Journal of Molecular Sciences, Vol. 20, Issue 14 https://doi.org/10.3390/ijms20143590	journal	July 2019
Evaluation of p63 and p73 antibodies for cross-reactivity Rosenbluth, Jennifer M.; Johnson, Kimberly; Tang, Luojia Cell Cycle, Vol. 8, Issue 22 https://doi.org/10.4161/cc.8.22.10036	journal	November 2009

Similar Records

Two oncogenes, v-fos and v-ras, cooperate to convert normal keratinocytes to squamous cell carcinoma

Journal Article · Sun Dec 31 23:00:00 EST 1989 · Proceedings of the National Academy of Sciences of the United States of America; (USA) · OSTI ID:6709918

Molecular Mechanisms of p63-Mediated Squamous Cancer Pathogenesis

Journal Article · Mon Jul 01 00:00:00 EDT 2019 · International Journal of Molecular Sciences (Online) · OSTI ID:1628384

Oncogenic Ras and ΔNp63α cooperate to recruit immunosuppressive polymorphonuclear myeloid-derived suppressor cells in a mouse model of squamous cancer pathogenesis

Citation Formats

References (61)

Similar Records

Related Subjects