Nanoparticle-enhanced proton beam immunoradiotherapy drives immune activation and durable tumor rejection

The combination of radiation therapy (RT) and immunotherapy has emerged as a promising treatment option in oncology. Historically, x-ray radiation (XRT) has been the most commonly used form of RT. However, proton beam therapy (PBT) is gaining recognition as a viable alternative, as it has been shown to produce similar outcomes to XRT while minimizing off-target effects. The effects of PBT on the antitumor immune response have only just begun to be described, and to our knowledge no studies to date have examined the effect of PBT as part of a combinatorial immunoradiotherapeutic strategy. Here, using a 2-tumor model of lung cancer in mice, we show that PBT in tandem with an anti-PD1 antibody substantially reduced growth in both irradiated and unirradiated tumors. This was accompanied by robust activation of the immune response, as evidenced by whole-tumor and single-cell RNA sequencing showing upregulation of a multitude of immune-related transcripts. This response was further significantly enhanced by the injection of the tumor to be irradiated with NBTXR3 nanoparticles. Tumors of mice treated with the triple combination exhibited increased infiltration and activation of cytotoxic immune cells. This triple combination eradicated both tumors in 37.5% of the treated mice and showed robust long-term immunity to cancer.

[1]  S. Paris,et al.  Combining a nanoparticle-mediated immunoradiotherapy with dual blockade of LAG3 and TIGIT improves the treatment efficacy in anti-PD1 resistant lung cancer , 2022, Journal of Nanobiotechnology.

[2]  S. Paris,et al.  Radiotherapy-activated NBTXR3 nanoparticles modulate cancer cell immunogenicity and TCR repertoire , 2022, Cancer Cell International.

[3]  F. Ginhoux,et al.  Tissue-resident FOLR2+ macrophages associate with CD8+ T cell infiltration in human breast cancer , 2022, Cell.

[4]  Z. Fridlender,et al.  The dual role of neutrophils in cancer. , 2021, Seminars in immunology.

[5]  J. Welsh,et al.  A radioenhancing nanoparticle mediated immunoradiation improves survival and generates long-term antitumor immune memory in an anti-PD1-resistant murine lung cancer model , 2021, Journal of Nanobiotechnology.

[6]  D. Nakae,et al.  Iron oxide nanoparticles exert inhibitory effects on N-Bis(2-hydroxypropyl)nitrosamine (DHPN)-induced lung tumorigenesis in rats. , 2021, Regulatory toxicology and pharmacology : RTP.

[7]  B. Johnston,et al.  The Current Landscape of NKT Cell Immunotherapy and the Hills Ahead , 2021, Cancers.

[8]  Steven H. Lin,et al.  Current Status and Application of Proton Therapy for Esophageal Cancer. , 2021, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[9]  J. Welsh,et al.  Radiotherapy enhanced by NBTXR3 nanoparticles overcomes anti-PD1 resistance and evokes abscopal effects. , 2021, International journal of radiation oncology, biology, physics.

[10]  C. Hedrick,et al.  Neutrophils in cancer: heterogeneous and multifaceted , 2021, Nature Reviews Immunology.

[11]  Cheng-song Sun,et al.  Innate lymphoid cells and cancer: Role in tumor progression and inhibition , 2021, European journal of immunology.

[12]  T. Seiwert,et al.  A phase I trial evaluating NBTXR3 activated by radiotherapy in combination with nivolumab or pembrolizumab in patients with advanced cancers. , 2021 .

[13]  R. Boidot,et al.  Impact of proton therapy on antitumor immune response , 2021, Scientific Reports.

[14]  W. Liang,et al.  Tumor-associated macrophage polarization promotes the progression of esophageal carcinoma , 2020, Aging.

[15]  Raphael Gottardo,et al.  Integrated analysis of multimodal single-cell data , 2020, Cell.

[16]  A. Mantovani,et al.  Neutrophil diversity and plasticity in tumour progression and therapy , 2020, Nature Reviews Cancer.

[17]  A. Kwong,et al.  Functional Implications of Cathelicidin Antimicrobial Protein in Breast Cancer and Tumor-Associated Macrophage Microenvironment , 2020, Biomolecules.

[18]  Feng-Sheng Wang,et al.  Epigenetic Regulation of Macrophage Marker Expression Profiles in Kawasaki Disease , 2020, Frontiers in Pediatrics.

[19]  M. Minopoli,et al.  Tumor Associated Neutrophils. Their Role in Tumorigenesis, Metastasis, Prognosis and Therapy , 2019, Front. Oncol..

[20]  M. Gennaro,et al.  Foam Cells: One Size Doesn't Fit All. , 2019, Trends in immunology.

[21]  B. Ryffel,et al.  CD3+ Macrophages Deliver Proinflammatory Cytokines by a CD3- and Transmembrane TNF-Dependent Pathway and Are Increased at the BCG-Infection Site , 2019, Front. Immunol..

[22]  S. Paris,et al.  DNA damage enhancement by radiotherapy-activated hafnium oxide nanoparticles improves cGAS-STING pathway activation in human colorectal cancer cells. , 2019, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[23]  R. Weinberg,et al.  Immuno-PET identifies the myeloid compartment as a key contributor to the outcome of the antitumor response under PD-1 blockade , 2019, Proceedings of the National Academy of Sciences.

[24]  S. Formenti,et al.  Radiation therapy to enhance tumor immunotherapy: a novel application for an established modality , 2019, International journal of radiation biology.

[25]  F. Ghiringhelli,et al.  Deciphering the Roles of Innate Lymphoid Cells in Cancer , 2019, Front. Immunol..

[26]  J. Sicklick,et al.  MST1R Kinase Accelerates Pancreatic Cancer Progression Via Effects on both Epithelial Cells and Macrophages , 2019, Oncogene.

[27]  K. Tretina,et al.  Interferon-induced guanylate-binding proteins: Guardians of host defense in health and disease , 2019, The Journal of experimental medicine.

[28]  M. Durante Proton beam therapy in Europe: more centres need more research , 2018, British Journal of Cancer.

[29]  D. Guan,et al.  Cannabinoid 2 receptor attenuates inflammation during skin wound healing by inhibiting M1 macrophages rather than activating M2 macrophages , 2018, Journal of Inflammation.

[30]  F. Hall,et al.  Exploiting Oncogenic Drivers along the CCNG1 Pathway for Cancer Therapy and Gene Therapy , 2018, Molecular therapy oncolytics.

[31]  J. Olson,et al.  Proton vs. Photon Radiation Therapy for Primary Gliomas: An Analysis of the National Cancer Data Base , 2018, Front. Oncol..

[32]  A. Butte,et al.  Reference-based analysis of lung single-cell sequencing reveals a transitional profibrotic macrophage , 2018, Nature Immunology.

[33]  W. Harmsen,et al.  A Comparison of Grade 4 Lymphopenia With Proton Versus Photon Radiation Therapy for Esophageal Cancer , 2019, Advances in radiation oncology.

[34]  Charlotte L. Scott,et al.  Macrophages and lipid metabolism , 2018, Cellular immunology.

[35]  Yi-xuan Yang,et al.  Inhibition of CD9 expression reduces the metastatic capacity of human hepatocellular carcinoma cell line MHCC97-H. , 2018, International journal of oncology.

[36]  B. S. Sørensen,et al.  "Radiobiology of Proton Therapy": Results of an international expert workshop. , 2018, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[37]  M. Shaul,et al.  Cancer‐related circulating and tumor‐associated neutrophils – subtypes, sources and function , 2018, The FEBS journal.

[38]  S. Chokshi,et al.  Immune checkpoint receptors: homeostatic regulators of immunity , 2018, Hepatology International.

[39]  R. Rengan,et al.  Proton beam therapy and immunotherapy: an emerging partnership for immune activation in non-small cell lung cancer. , 2018, Translational lung cancer research.

[40]  Fuqiang Pan,et al.  Distinct prognostic value of mRNA expression of guanylate-binding protein genes in skin cutaneous melanoma , 2018, Oncology letters.

[41]  I. Wistuba,et al.  VISTA is an inhibitory immune checkpoint that is increased after ipilimumab therapy in patients with prostate cancer , 2017, Nature Medicine.

[42]  A. M. Houghton,et al.  Neutrophils dominate the immune cell composition in non-small cell lung cancer , 2017, Nature Communications.

[43]  Yoshiyuki Hizukuri,et al.  Guanylate-binding protein 5 is a marker of interferon-γ-induced classically activated macrophages , 2016, Clinical & translational immunology.

[44]  K. Ricketts,et al.  Investigation into the effects of high-Z nano materials in proton therapy , 2016, Physics in medicine and biology.

[45]  Shohei Koyama,et al.  Adaptive resistance to therapeutic PD-1 blockade is associated with upregulation of alternative immune checkpoints , 2016, Nature Communications.

[46]  J. Bae,et al.  Tumor-Associated Macrophages and Neutrophils in Tumor Microenvironment , 2016, Mediators of inflammation.

[47]  Irene Garcia,et al.  Much More than M1 and M2 Macrophages, There are also CD169+ and TCR+ Macrophages , 2015, Front. Immunol..

[48]  M. Okoniewski,et al.  Complement is a central mediator of radiotherapy-induced tumor-specific immunity and clinical response. , 2015, Immunity.

[49]  Liyuan Zhang,et al.  Overexpression of CD9 correlates with tumor stage and lymph node metastasis in esophageal squamous cell carcinoma. , 2015, International journal of clinical and experimental pathology.

[50]  D. Corbeil,et al.  Tetraspanin CD9 determines invasiveness and tumorigenicity of human breast cancer cells , 2015, Oncotarget.

[51]  L. Levy,et al.  Phenotypic diversity and plasticity in circulating neutrophil subpopulations in cancer. , 2015, Cell reports.

[52]  Ben Tran,et al.  Prognostic role of neutrophil-to-lymphocyte ratio in solid tumors: a systematic review and meta-analysis. , 2014, Journal of the National Cancer Institute.

[53]  C. Garlanda,et al.  Tumor associated macrophages and neutrophils in cancer. , 2013, Immunobiology.

[54]  C. Garlanda,et al.  Tumor associated macrophages and neutrophils in tumor progression , 2013, Journal of cellular physiology.

[55]  Kihong Kim,et al.  Enhanced proton treatment in mouse tumors through proton irradiated nanoradiator effects on metallic nanoparticles , 2012, Physics in medicine and biology.

[56]  R. Palmqvist,et al.  The Distribution of Macrophages with a M1 or M2 Phenotype in Relation to Prognosis and the Molecular Characteristics of Colorectal Cancer , 2012, PloS one.

[57]  S. Tsuchida,et al.  Malignant Ovarian Tumors with Induced Expression of Carbonyl Reductase Show Spontaneous Regression , 2012, Clinical Medicine Insights. Oncology.

[58]  J. Albers,et al.  PLTP regulates STAT3 and NFκB in differentiated THP1 cells and human monocyte-derived macrophages. , 2011, Biochimica et biophysica acta.

[59]  A. Puig-Kröger,et al.  Activin A skews macrophage polarization by promoting a proinflammatory phenotype and inhibiting the acquisition of anti-inflammatory macrophage markers. , 2011, Blood.

[60]  G. Cheng,et al.  Polarization of tumor-associated neutrophil phenotype by TGF-beta: "N1" versus "N2" TAN. , 2009, Cancer cell.

[61]  Daxin Wang,et al.  Overexpressed PLTP in macrophage may promote cholesterol accumulation by prolonged endoplasmic reticulum stress. , 2017, Medical hypotheses.

[62]  D. Hou,et al.  Microarray-based determination of anti-inflammatory genes targeted by 6-(methylsulfinyl)hexyl isothiocyanate in macrophages. , 2010, Experimental and therapeutic medicine.