Publications

Featured Publications

Reticker-Flynn, N.E.**, Zhang, W., Belk, J.A., Basto, P.A., Escalante, N.K., Pilarowski, G.O.W., Bejnood, A., Martins, M.M., Kenkel, J.A., Linde, I.L., Bagchi, S., Yuan, R., Chang, S., Spitzer, M.H., Carmi, Y., Cheng, J., Tolentino, L.L., Choi, O., Wu, N., Kong, C., Gentles, A.J., Sunwoo, J.B., Satpathy, A.T., Plevritis, S.K., Engleman, E.G.**, “Lymph node colonization induces tumor-immune tolerance to promote distant metastasis.” Cell, May 2022. PMID: 35525247, DOI: 10.1016/j.cell.2022.04.019
** Co-corresponding authors
Highlighted in Nature Reviews CancerNature Reviews ImmunologyCancer DiscoveryScience SignalingTrends in CancerNCI Cancer Currents

Spitzer, M.H.*, Carmi, Y.*, Reticker-Flynn, N.E.*, Kwek, S.S., Madhireddy, D., Martins, M.M., Gherardini, P.F., Prestwood, T.R., Chabon, J., Bendall, S.C., Fong, L., Nolan, G.P.**, Engleman, E.G.** “Systemic immunity is required for effective cancer immunotherapy.” Cell, January 2017. PMID: 28111070
* Co-lead authors; ** Co-last authors


 

Reticker-Flynn, N.E. and Bhatia, S.N. "Aberrant glycosylation promotes lung cancer metastasis through adhesion to galectins in the metastatic niche." Cancer Discovery. February 2015. PMCID: PMC4367955.
Highlighted in Nature Reviews CancerCancer Discovery, and Nature

Reticker-Flynn, N.E., Braga Malta, D.F., Winslow, M.M., Lamar, J.M., Xu, M.J., Underhill, G.H., Hynes, R.O., Jacks, T.E., Bhatia, S.N. “A combinatorial extracellular matrix platform identifies cell-ECM interactions that correlate with metastasis.” Nature Communications. 3: October 9, 2012. PMCID: PMC3794716.

All Publications

Instructor, Otolaryngology — Head & Neck Surgery

Publications

  • Lymph nodes: at the intersection of cancer treatment and progression. Trends in cell biology Reticker-Flynn, N. E., Engleman, E. G. 2023

    Abstract

    Metastasis to lymph nodes (LNs) is a common feature of disease progression in most solid organ malignancies. Consequently, LN biopsy and lymphadenectomy are common clinical practices, not only because of their diagnostic utility but also as a means of deterring further metastatic spread. LN metastases have the potential to seed additional tissues and can induce metastatic tolerance, a process by which tumor-specific immune tolerance in LNs promotes further disease progression. Nonetheless, phylogenetic studies have revealed that distant metastases are not necessarily derived from nodal metastases. Furthermore, immunotherapy efficacy is increasingly being attributed to initiation of systemic immune responses within LNs. We argue that lymphadenectomy and nodal irradiation should be approached with caution, particularly in patients receiving immunotherapy.

    View details for DOI 10.1016/j.tcb.2023.04.001

    View details for PubMedID 37149414

  • Lymph node colonization induces tumor-immune tolerance to promote distant metastasis. Cell Reticker-Flynn, N. E., Zhang, W., Belk, J. A., Basto, P. A., Escalante, N. K., Pilarowski, G. O., Bejnood, A., Martins, M. M., Kenkel, J. A., Linde, I. L., Bagchi, S., Yuan, R., Chang, S., Spitzer, M. H., Carmi, Y., Cheng, J., Tolentino, L. L., Choi, O., Wu, N., Kong, C. S., Gentles, A. J., Sunwoo, J. B., Satpathy, A. T., Plevritis, S. K., Engleman, E. G. 2022

    Abstract

    For many solid malignancies, lymph node (LN) involvement represents a harbinger of distant metastatic disease and, therefore, an important prognostic factor. Beyond its utility as a biomarker, whether and how LN metastasis plays an active role in shaping distant metastasis remains an open question. Here, we develop a syngeneic melanoma mouse model of LN metastasis to investigate how tumors spread to LNs and whether LN colonization influences metastasis to distant tissues. We show that an epigenetically instilled tumor-intrinsic interferon response program confers enhanced LN metastatic potential by enabling the evasion of NK cells and promoting LN colonization. LN metastases resist T cell-mediated cytotoxicity, induce antigen-specific regulatory T cells, and generate tumor-specific immune tolerance that subsequently facilitates distant tumor colonization. These effects extend to human cancers and other murine cancer models, implicating a conserved systemic mechanism by which malignancies spread to distant organs.

    View details for DOI 10.1016/j.cell.2022.04.019

    View details for PubMedID 35525247

  • Cancer systems immunology. eLife Reticker-Flynn, N. E., Engleman, E. G. 2020; 9

    Abstract

    Tumor immunology is undergoing a renaissance due to the recent profound clinical successes of tumor immunotherapy. These advances have coincided with an exponential growth in the development of -omics technologies. Armed with these technologies and their associated computational and modeling toolsets, systems biologists have turned their attention to tumor immunology in an effort to understand the precise nature and consequences of interactions between tumors and the immune system. Such interactions are inherently multivariate, spanning multiple time and size scales, cell types, and organ systems, rendering systems biology approaches particularly amenable to their interrogation. While in its infancy, the field of 'Cancer Systems Immunology' has already influenced our understanding of tumor immunology and immunotherapy. As the field matures, studies will move beyond descriptive characterizations toward functional investigations of the emergent behavior that govern tumor-immune responses. Thus, Cancer Systems Immunology holds incredible promise to advance our ability to fight this disease.

    View details for DOI 10.7554/eLife.53839

    View details for PubMedID 32657757

  • Systemic Immunity Is Required for Effective Cancer Immunotherapy. Cell Spitzer, M. H., Carmi, Y., Reticker-Flynn, N. E., Kwek, S. S., Madhireddy, D., Martins, M. M., Gherardini, P. F., Prestwood, T. R., Chabon, J., Bendall, S. C., Fong, L., Nolan, G. P., Engleman, E. G. 2017; 168 (3): 487-502 e15

    Abstract

    Immune responses involve coordination across cell types and tissues. However, studies in cancer immunotherapy have focused heavily on local immune responses in the tumor microenvironment. To investigate immune activity more broadly, we performed an organism-wide study in genetically engineered cancer models using mass cytometry. We analyzed immune responses in several tissues after immunotherapy by developing intuitive models for visualizing single-cell data with statistical inference. Immune activation was evident in the tumor and systemically shortly after effective therapy was administered. However, during tumor rejection, only peripheral immune cells sustained their proliferation. This systemic response was coordinated across tissues and required for tumor eradication in several immunotherapy models. An emergent population of peripheral CD4 T cells conferred protection against new tumors and was significantly expanded in patients responding to immunotherapy. These studies demonstrate the critical impact of systemic immune responses that drive tumor rejection.

    View details for DOI 10.1016/j.cell.2016.12.022

    View details for PubMedID 28111070

  • Aberrant Glycosylation Promotes Lung Cancer Metastasis through Adhesion to Galectins in the Metastatic Niche CANCER DISCOVERY Reticker-Flynn, N. E., Bhatia, S. N. 2015; 5 (2): 168-181

    Abstract

    Metastasis is the leading cause of cancer-associated deaths. Although dissemination of tumor cells likely occurs early in tumorigenesis, the constituents of the microenvironment play essential rate-limiting roles in determining whether these cells will form clinically relevant tumors. Recent studies have uncovered many molecular factors that contribute to the establishment of a protumorigenic metastatic niche. Here, we demonstrate that galectin-3, whose expression has clinical associations with advanced malignancy and poor outcome, contributes to metastatic niche formation by binding to carbohydrates on metastatic cells. We show that galectin-3 is expressed early during tumorigenesis by both CD11b(+)Gr-1(+) and CD11b(+)Ly-6C(hi) leukocytes. Tumors mobilize these myeloid populations through secretion of soluble factors, including IL6. We find that metastatic cancer cells exhibit elevated presentation of the oncofetal galectin-3 carbohydrate ligand, the Thomsen-Friedenreich antigen, on their surfaces as a result of altered C2GnT2 and St6GalNAc4 glycosyltransferase activity that inhibits further glycosylation of this carbohydrate motif and promotes metastasis.Although clinical observations of elevated serum galectin-3 levels and altered glycosylation have been associated with malignancy, we identify novel roles for glycosyltransferases in promoting adhesion to galectins in the metastatic niche. This identification of a cytokine-leukocyte-glycosylation axis in metastasis provides mechanistic explanations for clinical associations between malignancy and aberrant glycosylation.

    View details for DOI 10.1158/2159-8290.CD-13-0760

    View details for Web of Science ID 000349393600025

    View details for PubMedID 25421439

    View details for PubMedCentralID PMC4367955

  • A combinatorial extracellular matrix platform identifies cell-extracellular matrix interactions that correlate with metastasis NATURE COMMUNICATIONS Reticker-Flynn, N. E., Malta, D. F., Winslow, M. M., Lamar, J. M., Xu, M. J., Underhill, G. H., Hynes, R. O., Jacks, T. E., Bhatia, S. N. 2012; 3

    Abstract

    Extracellular matrix interactions have essential roles in normal physiology and many pathological processes. Although the importance of extracellular matrix interactions in metastasis is well documented, systematic approaches to identify their roles in distinct stages of tumorigenesis have not been described. Here we report a novel-screening platform capable of measuring phenotypic responses to combinations of extracellular matrix molecules. Using a genetic mouse model of lung adenocarcinoma, we measure the extracellular matrix-dependent adhesion of tumour-derived cells. Hierarchical clustering of the adhesion profiles differentiates metastatic cell lines from primary tumour lines. Furthermore, we uncovered that metastatic cells selectively associate with fibronectin when in combination with galectin-3, galectin-8 or laminin. We show that these molecules correlate with human disease and that their interactions are mediated in part by α3β1 integrin. Thus, our platform allowed us to interrogate interactions between metastatic cells and their microenvironments, and identified extracellular matrix and integrin interactions that could serve as therapeutic targets.

    View details for DOI 10.1038/ncomms2128

    View details for Web of Science ID 000313514100032

    View details for PubMedID 23047680

  • Interrogating the roles of lymph node metastasis in systemic immune surveillance. Clinical & experimental metastasis Basto, P. A., Reticker-Flynn, N. E. 2024

    Abstract

    Lymph nodes (LNs) are principal orchestrators of the adaptive immune response, yet in the context of malignancy, they are typically the first sites of metastasis. When tumors spread to LNs, they alter the immune repertoire, ultimately reconditioning it in a manner that suppresses anti-tumor immunity and promotes further metastatic dissemination. Conversely, activation of anti-tumor immunity within LNs is essential for immunotherapy, suggesting clinical approaches to radiotherapy in LNs and lymphadenectomy may need to be reconsidered in the context of immune checkpoint blockade (ICB). Herein, we discuss our understanding of the immune remodeling that coincides with LN metastasis as well as recent clinical studies exploring neoadjuvant immunotherapy and the roles of LNs in treatment of solid organ malignancies.

    View details for DOI 10.1007/s10585-023-10261-3

    View details for PubMedID 38315348

    View details for PubMedCentralID 5998822

  • Neutrophil-activating therapy for the treatment of cancer. Cancer cell Linde, I. L., Prestwood, T. R., Qiu, J., Pilarowski, G., Linde, M. H., Zhang, X., Shen, L., Reticker-Flynn, N. E., Chiu, D. K., Sheu, L. Y., Van Deursen, S., Tolentino, L. L., Song, W., Engleman, E. G. 2023

    Abstract

    Despite their cytotoxic capacity, neutrophils are often co-opted by cancers to promote immunosuppression, tumor growth, and metastasis. Consequently, these cells have received little attention as potential cancer immunotherapeutic agents. Here, we demonstrate in mouse models that neutrophils can be harnessed to induce eradication of tumors and reduce metastatic seeding through the combined actions of tumor necrosis factor, CD40 agonist, and tumor-binding antibody. The same combination activates human neutrophils invitro, enabling their lysis of human tumor cells. Mechanistically, this therapy induces rapid mobilization and tumor infiltration of neutrophils along with complement activation in tumors. Complement component C5a activates neutrophils to produce leukotriene B4, which stimulates reactive oxygen species production via xanthine oxidase, resulting in oxidative damage and Tcell-independent clearance of multiple tumor types. These data establish neutrophils as potent anti-tumor immune mediators and define an inflammatory pathway that can be harnessed to drive neutrophil-mediated eradication of cancer.

    View details for DOI 10.1016/j.ccell.2023.01.002

    View details for PubMedID 36706760

  • Transient cell-in-cell formation underlies tumor relapse and resistance to immunotherapy. eLife Gutwillig, A., Santana-Magal, N., Farhat-Younis, L., Rasoulouniriana, D., Madi, A., Luxenburg, C., Cohen, J., Padmanabhan, K., Shomron, N., Shapira, G., Gleiberman, A., Parikh, R., Levy, C., Feinmesser, M., Hershkovitz, D., Zemser-Werner, V., Zlotnik, O., Kroon, S., Hardt, W., Debets, R., Reticker-Flynn, N. E., Rider, P., Carmi, Y. 2022; 11

    Abstract

    Despite the remarkable successes of cancer immunotherapies, the majority of patients will experience only partial response followed by relapse of resistant tumors. While treatment resistance has frequently been attributed to clonal selection and immunoediting, comparisons of paired primary and relapsed tumors in melanoma and breast cancers indicate that they share the majority of clones. Here, we demonstrate in both mouse models and clinical human samples that tumor cells evade immunotherapy by generating unique transient cell-in-cell structures, which are resistant to killing by T cells and chemotherapies. While the outer cells in this cell-in-cell formation are often killed by reactive T cells, the inner cells remain intact and disseminate into single tumor cells once T cells are no longer present. This formation is mediated predominantly by IFNgamma-activated T cells, which subsequently induce phosphorylation of the transcription factors signal transducer and activator of transcription 3 (STAT3) and early growth response-1 (EGR-1) in tumor cells. Indeed, inhibiting these factors prior to immunotherapy significantly improves its therapeutic efficacy. Overall, this work highlights a currently insurmountable limitation of immunotherapy and reveals a previously unknown resistance mechanism which enables tumor cells to survive immune-mediated killing without altering their immunogenicity.

    View details for DOI 10.7554/eLife.80315

    View details for PubMedID 36124553

  • Identification of cell types in multiplexed in situ images by combining protein expression and spatial information using CELESTA. Nature methods Zhang, W., Li, I., Reticker-Flynn, N. E., Good, Z., Chang, S., Samusik, N., Saumyaa, S., Li, Y., Zhou, X., Liang, R., Kong, C. S., Le, Q., Gentles, A. J., Sunwoo, J. B., Nolan, G. P., Engleman, E. G., Plevritis, S. K. 2022

    Abstract

    Advances in multiplexed in situ imaging are revealing important insights in spatial biology. However, cell type identification remains a major challenge in imaging analysis, with most existing methods involving substantial manual assessment and subjective decisions for thousands of cells. We developed an unsupervised machine learning algorithm, CELESTA, which identifies the cell type of each cell, individually, using the cell's marker expression profile and, when needed, its spatial information. We demonstrate the performance of CELESTA on multiplexed immunofluorescence images of colorectal cancer and head and neck squamous cell carcinoma (HNSCC). Using the cell types identified by CELESTA, we identify tissue architecture associated with lymph node metastasis in HNSCC, and validate our findings in an independent cohort. By coupling our spatial analysis with single-cell RNA-sequencing data on proximal sections of the same specimens, we identify cell-cell crosstalk associated with lymph node metastasis, demonstrating the power of CELESTA to facilitate identification of clinically relevant interactions.

    View details for DOI 10.1038/s41592-022-01498-z

    View details for PubMedID 35654951