bluuebunny/crossref_metadata_2025 huggingface dataset:
Data from Cross-Talk between Myeloid-Derived Suppressor Cells and Mast Cells Mediates Tumor-Specific Immunosuppression in Prostate Cancer
Authors: Jachetti Elena , Cancila Valeria , Rigoni Alice , Bongiovanni Lucia , Cappetti Barbara , Belmonte Beatrice , Enriquez Claudia , Casalini Patrizia , Ostano Paola , Frossi Barbara , Sangaletti Sabina , Chiodoni Claudia , Chiorino Giovanna , Pucillo Carlo E. , Tripodo Claudio , Colombo Mario P.
Published: April 2023

Abstract
Immunotherapy, including the use of checkpoint inhibitors, is a potent therapeutic approach for some cancers, but has limited success with prostate tumors, in which immune suppression is instigated by the tumor. The immunosuppressive capacity of mast cells, which promote adenocarcinoma development in the prostate, prompted our investigation on whether mast cells promote tolerance to SV40 Large-T antigen, the transforming oncogene in transgenic adenocarcinoma of the mouse prostate (TRAMP) mice. The incidence of adenocarcinoma was reduced in the offspring of a cross between TRAMP mice and mast cell–deficient KitWsh mice. TRAMP mice are tolerant to the SV40 Large T antigen, which is otherwise immunogenic in normal syngeneic B6 mice. Genetic ablation of mast cells in TRAMP mice restored their ability to mount a tumor-specific cytotoxic T-cell response. In KitWsh-TRAMP mice, the restored T-cell immunity correlated with the reduced activity of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC), along with their reduced expression of Arg1, Nos2, and Stat3. Having found that CD40L-expressing mast cells can interact in vivo with CD40-expressing PMN-MDSC, we then determined that only KitWsh-TRAMP mice reconstituted with mast cells expressing CD40L could restore PMN-MDSCs suppressive functions, T-cell unresponsiveness and adenocarcinoma development. Thus, mast cells have an immunoregulatory effect on PMN-MDSCs activity through CD40L-CD40 interaction, favoring immunosuppression and tumor onset. In prostate cancer patients, in silico analyses correlated poor clinical outcomes with high expression of genes related to mast cells and PMN-MDSCs. Cancer Immunol Res; 6(5); 552–65. ©2018 AACR.
Data from T Cells Specific for an Unconventional Natural Antigen Fail to Recognize Leukemic Cells
Authors: Pont Margot J. , Oostvogels Rimke , van Bergen Cornelis A.M. , van der Meijden Edith D. , Honders Maria W. , Bliss Sophie , Jongsma Marlieke L.M. , Lokhorst Henk M. , Falkenburg J.H. Frederik , Mutis Tuna , Griffioen Marieke , Spaapen Robbert M.
Published: April 2023

Abstract
MHC-bound peptides from aberrant proteins may be a specific immunotherapeutic target on cancer cells. Because of difficulties in identifying such antigens, viral or model antigens have so far been used to study their biological relevance. We here identify a naturally existing human T-cell epitope derived from a truncated protein. The antigenic peptide is derived from the gene TTK only through an alternative transcript containing a premature termination codon that may target the transcript for nonsense-mediated decay (NMD). This antigen is recognized by HLA-A*02:01–restricted CD8+ T cells derived from an allotransplanted leukemia patient. Functional analyses showed that these T cells failed to recognize several HLA-matched primary leukemic cells that expressed the alternative TTK transcript. Conventional antigen processing and presentation were not affected, suggesting that leukemic cells modify the generation of antigens processed from aberrant proteins. This natural TTK epitope provides insights in the source of transcripts producing antigenic epitopes in healthy and leukemic cells. Our data underscore potential pitfalls of targeting NMD-derived or other unconventionally generated epitopes as immunotherapeutic approach.
Data from Chronic Lymphocytic Leukemia Monitoring with a Lamprey Idiotope-Specific Antibody
Authors: Nakahara Hirotomo , Herrin Brantley R. , Alder Matthew N. , Catera Rosa , Yan Xiao-Jie , Chiorazzi Nicholas , Cooper Max D.
Published: April 2023

Abstract
For antigen recognition, lampreys use leucine-rich repeats (LRR) instead of immunoglobulin V-(D)-J domains to generate variable lymphocyte receptors (VLR) of three types: VLRA, VLRB, and VLRC. VLRB-bearing lymphocytes respond to immunization with proliferation and differentiation into plasmacytes that secrete multivalent VLRB antibodies. Here, we immunized lampreys with B cells from patients with chronic lymphocytic leukemia (CLL) to generate recombinant monoclonal VLRB antibodies, one of which, VLR39, was specific for the donor CLL cells. The target epitope of VLR39 was shown to be the complementarity determining region 3 (CDR3) of the heavy chain variable region (VH) of the B-cell receptor. Using this antibody to monitor the CLL donor after chemoimmunotherapy-induced remission, we detected VLR39+ B cells in the patient 51 months later, before significant increase in lymphocyte count or CD5+ B cells. This indication of reemergence of the leukemic clone was verified by VH sequencing. Lamprey antibodies can exhibit exquisite specificity for a protein epitope, a CLL signature VH CDR3 sequence in this case, and offer a rapid strategy for generating anti-idiotype antibodies for early detection of leukemia recurrence. Cancer Immunol Res; 1(4); 223–8. ©2013 AACR.
Data from Immune-Checkpoint Blockade Opposes CD8+ T-cell Suppression in Human and Murine Cancer
Authors: Pfannenstiel Lukas W. , Diaz-Montero C. Marcela , Tian Ye F. , Scharpf Joseph , Ko Jennifer S. , Gastman Brian R.
Published: April 2023

Abstract
Immune-checkpoint blockade enhances antitumor responses against cancers. One cancer type that is sensitive to checkpoint blockade is squamous cell carcinoma of the head and neck (SCCHN), which we use here to study limitations of this treatment modality. We observed that CD8+ tumor-infiltrating lymphocytes (TILs) in SCCHN and melanoma express excess immune checkpoints components PD-1 and Tim-3 and are also CD27−/CD28−, a phenotype we previously associated with immune dysfunction and suppression. In ex vivo experiments, patients' CD8+ TILs with this phenotype suppressed proliferation of autologous peripheral blood T cells. Similar phenotype and function of TILs was observed in the TC-1 mouse tumor model. Treatment of TC-1 tumors with anti–PD-1 or anti–Tim-3 slowed tumor growth in vivo and reversed the suppressive function of multi-checkpoint+ CD8+ TIL. Similarly, treatment of both human and mouse PD-1+ Tim-3+ CD8+ TILs with anticheckpoint antibodies ex vivo reversed their suppressive function. These suppressive CD8+ TILs from mice and humans expressed ligands for PD-1 and Tim-3 and exerted their suppressive function via IL10 and close contact. To model therapeutic strategies, we combined anti–PD-1 blockade with IL7 cytokine therapy or with transfer of antigen-specific T cells. Both strategies resulted in synergistic antitumor effects and reduced suppressor cell function. These findings enhance our understanding of checkpoint blockade in cancer treatment and identify strategies to promote synergistic activities in the context of other immunotherapies.
Data from Wnt3a/β-Catenin Signaling Conditions Differentiation of Partially Exhausted T-effector Cells in Human Cancers
Authors: Schinzari Valeria , Timperi Eleonora , Pecora Giulia , Palmucci Francesco , Gallerano Daniela , Grimaldi Alessio , Covino Daniela Angela , Guglielmo Nicola , Melandro Fabio , Manzi Emy , Sagnotta Andrea , Lancellotti Francesco , Sacco Luca , Chirletti Piero , Grazi Gian Luca , Rossi Massimo , Barnaba Vincenzo
Published: April 2023

Abstract
In this study, we investigated the role of the Wnt/β-catenin signaling pathway in antitumor immune responses. We report that the concentration of secreted Wnt3a was significantly higher in conditioned medium from tumor or nontumor tissues obtained from all hepatocellular carcinoma or colorectal cancer patients tested, than in serum of healthy donors or patients. In addition, both Wnt3a and β-catenin were overexpressed by tumor-infiltrating and nontumor-infiltrating CD4+ or CD8+ T cells. The majority of these T cells expressed a dysfunctional effector memory Eomes+T-bet−phenotype that we defined as partially exhausted, because they performed effector functions (in terms of interferon-γ and tumor necrosis factor-α production, as well as CD107a mobilization) despite their PD-1 expression. Wnt3a/β-catenin signaling in T naïve cells in vitro recapitulated the T-cell setting in vivo. Indeed, the differentiation of cultured T naïve cells was arrested, producing cells that resembled the EomeshighT-betlowβ-cateninhigh T cells with moderate effector functions that infiltrated tumor and nontumor areas. Wnt3a blockade improved the capacity of T naïve cells to differentiate into effector cells in vitro. However, Wnt3a blockade did not affect the function and phenotype of differentiated, partially exhausted, tumor-infiltrating T cells ex vivo. Taken together, our data suggest that Wnt3a blockade halts the capacity of Wnt/β-catenin signaling to inhibit the differentiation of T naïve cells, but it does not restore the dysfunction of differentiated T cells, in the tumor setting. Cancer Immunol Res; 6(8); 941–52. ©2018 AACR.
Data from IAP Antagonists Enhance Cytokine Production from Mouse and Human iNKT Cells
Authors: Clancy-Thompson Eleanor , Ali Lestat , Bruck Patrick T. , Exley Mark A. , Blumberg Richard S. , Dranoff Glenn , Dougan Michael , Dougan Stephanie K.
Published: April 2023

Abstract
Inhibitor of apoptosis protein (IAP) antagonists are in clinical trials for a variety of cancers, and mouse models show synergism between IAP antagonists and anti–PD-1 immunotherapy. Although IAP antagonists affect the intrinsic signaling of tumor cells, their most pronounced effects are on immune cells and the generation of antitumor immunity. Here, we examined the effects of IAP antagonism on T-cell development using mouse fetal thymic organ culture and observed a selective loss of iNKT cells, an effector cell type of potential importance for cancer immunotherapy. Thymic iNKT-cell development probably failed due to increased strength of TCR signal leading to negative selection, given that mature iNKT cells treated with IAP antagonists were not depleted, but had enhanced cytokine production in both mouse and human ex vivo cultures. Consistent with this, mature mouse primary iNKT cells and iNKT hybridomas increased production of effector cytokines in the presence of IAP antagonists. In vivo administration of IAP antagonists and α-GalCer resulted in increased IFNγ and IL-2 production from iNKT cells and decreased tumor burden in a mouse model of melanoma lung metastasis. Human iNKT cells also proliferated and increased IFNγ production dramatically in the presence of IAP antagonists, demonstrating the utility of these compounds in adoptive therapy of iNKT cells. Cancer Immunol Res; 6(1); 25–35. ©2017 AACR.
Data from IL13-Mediated Dectin-1 and Mannose Receptor Overexpression Promotes Macrophage Antitumor Activities through Recognition of Sialylated Tumor Cells
Authors: Alaeddine Mohamad , Prat Mélissa , Poinsot Véréna , Gouazé-Andersson Valérie , Authier Hélène , Meunier Etienne , Lefèvre Lise , Alric Camille , Dardenne Christophe , Bernad José , Alric Laurent , Segui Bruno , Balard Patricia , Couderc François , Couderc Bettina , Pipy Bernard , Coste Agnès
Published: April 2023

Abstract
Macrophage-mediated cytotoxicity is controlled by surface receptor expression and activation. Despite the numerous studies documenting the role of macrophage C-type lectin receptors (CLR) in pathogen elimination, little is known about their contribution to antitumor responses. Here, we report that IL13 inhibits T-cell lymphoma and ovarian adenocarcinoma development in tumor-bearing mice through the conversion of tumor-supporting macrophages to cytotoxic effectors, characterized by a CLR signature composed of dectin-1 and mannose receptor (MR). We show that dectin-1 and MR are critical for the recognition of tumor cells through sialic acid–specific glycan structure on their surface and for the subsequent activation of macrophage tumoricidal response. Finally, we validated that IL13 antitumor effect mediated by dectin-1 and MR overexpression on macrophages can extend to various types of human tumors. Therefore, these results identify these CLRs as potential targets to promote macrophage antitumor response and represent an attractive approach to elicit tumor-associated macrophage tumoricidal properties.
Data from Autocrine TGFβ Is a Survival Factor for Monocytes and Drives Immunosuppressive Lineage Commitment
Authors: Gonzalez-Junca Alba , Driscoll Kyla E. , Pellicciotta Ilenia , Du Shisuo , Lo Chen Hao , Roy Ritu , Parry Renate , Tenvooren Iliana , Marquez Diana M. , Spitzer Matthew H. , Barcellos-Hoff Mary Helen
Published: April 2023

Abstract
Transforming growth factor β (TGFβ) is an effector of immune suppression and contributes to a permissive tumor microenvironment that compromises effective immunotherapy. We identified a correlation between TGFB1 and genes expressed by myeloid cells, but not granulocytes, in The Cancer Genome Atlas lung adenocarcinoma data, in which high TGFB1 expression was associated with poor survival. To determine whether TGFβ affected cell fate decisions and lineage commitment, we studied primary cultures of CD14+ monocytes isolated from peripheral blood of healthy donors. We discovered that TGFβ was a survival factor for CD14+ monocytes, which rapidly executed an apoptotic program in its absence. Continued exposure to TGFβ in combination with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 6 (IL6) amplified HLA-DRlowCD14+CD11b+CD33+ myeloid-derived suppressor cells (MDSCs) at the expense of macrophage and dendritic cell (DC) differentiation. MDSCs generated in the presence of TGFβ were more effective in suppressing T-cell proliferation and promoted the T regulatory cell phenotype. In contrast, inhibition of TGFβ signaling using a small-molecule inhibitor of receptor kinase activity in CD14+ monocytes treated with GM-CSF and IL6 decreased MDSC differentiation and increased differentiation to proinflammatory macrophages and antigen-presenting DCs. The effect of autocrine and paracrine TGFβ on myeloid cell survival and lineage commitment suggests that pharmacologic inhibition of TGFβ-dependent signaling in cancer would favor antitumor immunity.
Data from Treatment-Related Adverse Events Predict Improved Clinical Outcome in NSCLC Patients on KEYNOTE-001 at a Single Center
Authors: Lisberg Aaron , Tucker D. Andrew , Goldman Jonathan W. , Wolf Brian , Carroll James , Hardy Ariana , Morris Karolyn , Linares Paulina , Adame Carlos , Spiegel Marshall L. , Wells Courtney , McKenzie Jordan , Ledezma Blanca , Mendenhall Melody , Abarca Phillip , Bornazyan Krikor , Hunt Jaime , Moghadam Nima , Chong Natalie , Nameth Danielle , Marx Caitlin , Madrigal John , Vangala Sitaram , Shaverdian Narek , Elashoff David , Garon Edward B.
Published: April 2023

Abstract
We retrospectively analyzed non–small cell lung cancer (NSCLC) patients from a single center treated with pembrolizumab on the KEYNOTE-001 trial and evaluated the association between treatment-related adverse events (trAEs) and clinical outcomes. Investigators reported AEs on trial and graded them according to Common Terminology Criteria for Adverse Events v4.0, labeling them as unlikely, possibly, or probably treatment-related. AEs labeled as possibly/probably related were considered trAEs for this analysis. The relationship between the incidence of a trAE and clinical outcomes was evaluated. Ninety-seven NSCLC patients treated on KEYNOTE-001 at the University of California, Los Angeles were evaluated. Ten percent (85/826) of AEs were trAEs, occurring in 40% (39/97) of patients. The most frequent trAEs were rash (21% patients), fatigue (6% patients), and hypothyroidism (6% patients). The 39 patients that experienced a trAE had increased objective response rate (ORR, 38.5%), progression-free survival (PFS: median, 248 days), and overall survival (OS: median, 493 days), compared with the 58 patients that did not (ORR: 8.9%, PFS: median 60 days, OS: median 144.5 days). The observed association between trAEs and improved clinical outcome persisted when using Cox proportional hazards regression models to assess the confounding effect of covariates and mitigate guarantee-time bias. The association also remained when data were substratified by grade, degree of association, and treatment-related select AE designation. This single-center analysis revealed that trAEs predicted for improved clinical outcome with pembrolizumab, and when controlling for guarantee-time bias and plausible confounders, this association remained. This observed relationship adds to our understanding of anti–PD-1 therapy and could aid clinicians in identifying patients most likely to benefit from therapy. Cancer Immunol Res; 6(3); 288–94. ©2018 AACR.
Data from Exosomes Shuttle TREX1-Sensitive IFN-Stimulatory dsDNA from Irradiated Cancer Cells to DCs
Authors: Diamond Julie M. , Vanpouille-Box Claire , Spada Sheila , Rudqvist Nils-Petter , Chapman Jessica R. , Ueberheide Beatrix M. , Pilones Karsten A. , Sarfraz Yasmeen , Formenti Silvia C. , Demaria Sandra
Published: April 2023

Abstract
Radiotherapy (RT) used at immunogenic doses leads to accumulation of cytosolic double-stranded DNA (dsDNA) in cancer cells, which activates type I IFN (IFN-I) via the cGAS/STING pathway. Cancer cell–derived IFN-I is required to recruit BATF3-dependent dendritic cells (DC) to poorly immunogenic tumors and trigger antitumor T-cell responses in combination with immune checkpoint blockade. We have previously demonstrated that the exonuclease TREX1 regulates radiation immunogenicity by degrading cytosolic dsDNA. Tumor-derived DNA can also activate cGAS/STING-mediated production of IFN-I by DCs infiltrating immunogenic tumors. However, how DNA from cancer cells is transferred to the cytoplasm of DCs remains unclear. Here, we showed that tumor-derived exosomes (TEX) produced by irradiated mouse breast cancer cells (RT-TEX) transfer dsDNA to DCs and stimulate DC upregulation of costimulatory molecules and STING-dependent activation of IFN-I. In vivo, RT-TEX elicited tumor-specific CD8+ T-cell responses and protected mice from tumor development significantly better than TEX from untreated cancer cells in a prophylactic vaccination experiment. We demonstrated that the IFN-stimulatory dsDNA cargo of RT-TEX is regulated by TREX1 expression in the parent cells. Overall, these results identify RT-TEX as a mechanism whereby IFN-stimulatory dsDNA is transferred from irradiated cancer cells to DCs. We have previously shown that the expression of TREX1 is dependent on the RT dose size. Thus, these data have important implications for the use of RT with immunotherapy. Cancer Immunol Res; 6(8); 910–20. ©2018 AACR.
Data from Reducing Ex Vivo Culture Improves the Antileukemic Activity of Chimeric Antigen Receptor (CAR) T Cells
Authors: Ghassemi Saba , Nunez-Cruz Selene , O'Connor Roddy S. , Fraietta Joseph A. , Patel Prachi R. , Scholler John , Barrett David M. , Lundh Stefan M. , Davis Megan M. , Bedoya Felipe , Zhang Changfeng , Leferovich John , Lacey Simon F. , Levine Bruce L. , Grupp Stephan A. , June Carl H. , Melenhorst J. Joseph , Milone Michael C.
Published: April 2023

Abstract
The success of chimeric antigen receptor (CAR)–mediated immunotherapy in acute lymphoblastic leukemia (ALL) highlights the potential of T-cell therapies with directed cytotoxicity against specific tumor antigens. The efficacy of CAR T-cell therapy depends on the engraftment and persistence of T cells following adoptive transfer. Most protocols for T-cell engineering routinely expand T cells ex vivo for 9 to 14 days. Because the potential for engraftment and persistence is related to the state of T-cell differentiation, we hypothesized that reducing the duration of ex vivo culture would limit differentiation and enhance the efficacy of CAR T-cell therapy. We demonstrated that T cells with a CAR-targeting CD19 (CART19) exhibited less differentiation and enhanced effector function in vitro when harvested from cultures at earlier (day 3 or 5) compared with later (day 9) timepoints. We then compared the therapeutic potential of early versus late harvested CART19 in a murine xenograft model of ALL and showed that the antileukemic activity inversely correlated with ex vivo culture time: day 3 harvested cells showed robust tumor control despite using a 6-fold lower dose of CART19, whereas day 9 cells failed to control leukemia at limited cell doses. We also demonstrated the feasibility of an abbreviated culture in a large-scale current good manufacturing practice–compliant process. Limiting the interval between T-cell isolation and CAR treatment is critical for patients with rapidly progressing disease. Generating CAR T cells in less time also improves potency, which is central to the effectiveness of these therapies. Cancer Immunol Res; 6(9); 1100–9. ©2018 AACR.
Data from A CD40 Agonist and PD-1 Antagonist Antibody Reprogram the Microenvironment of Nonimmunogenic Tumors to Allow T-cell–Mediated Anticancer Activity
Authors: Ma Hayley S. , Poudel Bibhav , Torres Evanthia Roussos , Sidhom John-William , Robinson Tara M. , Christmas Brian , Scott Blake , Cruz Kayla , Woolman Skylar , Wall Valerie Z. , Armstrong Todd , Jaffee Elizabeth M.
Published: April 2023

Abstract
In cancers with tumor-infiltrating lymphocytes (TILs), monoclonal antibodies (mAbs) that block immune checkpoints such as CTLA-4 and PD-1/PD-L1 promote antitumor T-cell immunity. Unfortunately, most cancers fail to respond to single-agent immunotherapies. T regulatory cells, myeloid derived suppressor cells (MDSCs), and extensive stromal networks within the tumor microenvironment (TME) dampen antitumor immune responses by preventing T-cell infiltration and/or activation. Few studies have explored combinations of immune-checkpoint antibodies that target multiple suppressive cell populations within the TME, and fewer have studied the combinations of both agonist and antagonist mAbs on changes within the TME. Here, we test the hypothesis that combining a T-cell–inducing vaccine with both a PD-1 antagonist and CD40 agonist mAbs (triple therapy) will induce T-cell priming and TIL activation in mouse models of nonimmunogenic solid malignancies. In an orthotopic breast cancer model and both subcutaneous and metastatic pancreatic cancer mouse models, only triple therapy was able to eradicate most tumors. The survival benefit was accompanied by significant tumor infiltration of IFNγ-, Granzyme B-, and TNFα-secreting effector T cells. Further characterization of immune populations was carried out by high-dimensional flow-cytometric clustering analysis and visualized by t-distributed stochastic neighbor embedding (t-SNE). Triple therapy also resulted in increased infiltration of dendritic cells, maturation of antigen-presenting cells, and a significant decrease in granulocytic MDSCs. These studies reveal that combination CD40 agonist and PD-1 antagonist mAbs reprogram immune resistant tumors in favor of antitumor immunity.
Data from The Atypical Receptor CCRL2 Is Essential for Lung Cancer Immune Surveillance
Authors: Del Prete Annalisa , Sozio Francesca , Schioppa Tiziana , Ponzetta Andrea , Vermi William , Calza Stefano , Bugatti Mattia , Salvi Valentina , Bernardini Giovanni , Benvenuti Federica , Vecchi Annunciata , Bottazzi Barbara , Mantovani Alberto , Sozzani Silvano
Published: April 2023

Abstract
CCRL2 is a nonsignaling seven-transmembrane domain receptor. CCRL2 binds chemerin, a protein that promotes chemotaxis of leukocytes, including macrophages and natural killer (NK) cells. In addition, CCRL2 controls the inflammatory response in different pathologic settings, such as hypersensitivity, inflammatory arthritis, and experimental autoimmune encephalitis. Here, we investigated the role of CCRL2 in the regulation of lung cancer–related inflammation. The genetic deletion of Ccrl2 promoted tumor progression in urethane-induced and in KrasG12D/+/p53LoxP lung tumor mouse models. Similarly, a Kras-mutant lung tumor displayed enhanced growth in Ccrl2-deficient mice. This phenotype was associated with a reduced inflammatory infiltrate characterized by the impaired recruitment of several leukocyte populations including NK cells. Bone marrow chimeras showed that CCRL2 expression by the nonhematopoietic cell compartment was responsible for the increased tumor formation observed in Kras-mutant Ccrl2-deficient mice. In human and mouse lungs, CCRL2 was expressed by a fraction of CD31+ endothelial cells, where it could control NK infiltration. Elevated CCRL2 expression in biopsies from human lung adenocarcinoma positively correlated with clinical outcome. These results provide evidence for a crucial role of CCRL2 in shaping an anti–lung tumor immune response.
Data from Coexpression of Inhibitory Receptors Enriches for Activated and Functional CD8+ T Cells in Murine Syngeneic Tumor Models
Authors: Xiong Huizhong , Mittman Stephanie , Rodriguez Ryan , Pacheco-Sanchez Patricia , Moskalenko Marina , Yang Yagai , Elstrott Justin , Ritter Alex T. , Müller Sören , Nickles Dorothee , Arenzana Teresita L. , Capietto Aude-Hélène , Delamarre Lélia , Modrusan Zora , Rutz Sascha , Mellman Ira , Cubas Rafael
Published: April 2023

Abstract
Exhausted T cells have been described in cancer patients and murine tumor models largely based on their expression of various inhibitory receptors. Understanding of the functional attributes of these cells is limited. Here, we report that among CD8+ T cells in commonly used syngeneic tumor models, the coexpression of inhibitory receptors PD-1, LAG3, and TIM3 defined a group of highly activated and functional effector cells. Coexpression of these receptors further enriched for antigen-specific cells with increased T-cell receptor clonality. Anti–PD-L1 treatment increased the number and activation of these triple-positive CD8+ T cells without affecting the density of PD-1− cells. The intratumoral density of CD8+ T cells coexpressing inhibitory receptors negatively correlated with tumor burden. The density ratio and pretreatment phenotype of CD8+ T cells coexpressing inhibitory receptors was positively correlated with response across a variety of tumor models. Our results demonstrate that coexpression of inhibitory receptors is not a signifier of exhausted T cells, but rather can define a group of activated and functional effector cells in syngeneic tumor models. In the cancer setting, these cells could represent a heterogeneous population of not only exhausted but also highly activated cells responsive to treatment.
Data from Cancer-Associated Fibroblasts Promote Immunosuppression by Inducing ROS-Generating Monocytic MDSCs in Lung Squamous Cell Carcinoma
Authors: Xiang Handan , Ramil Carlo P. , Hai Josephine , Zhang Chunsheng , Wang Huijun , Watkins Amanda A. , Afshar Roshi , Georgiev Peter , Sze Marc A. , Song Xuelei S. , Curran Patrick J. , Cheng Mangeng , Miller J. Richard , Sun Dongyu , Loboda Andrey , Jia Yanlin , Moy Lily Y. , Chi An , Brandish Philip E.
Published: April 2023

Abstract
Cancer-associated fibroblasts (CAF) represent a functionally heterogeneous population of activated fibroblasts that constitutes a major component of tumor stroma. Although CAFs have been shown to promote tumor growth and mediate resistance to chemotherapy, the mechanisms by which they may contribute to immune suppression within the tumor microenvironment (TME) in lung squamous cell carcinoma (LSCC) remain largely unexplored. Here, we identified a positive correlation between CAF and monocytic myeloid cell abundances in 501 primary LSCCs by mining The Cancer Genome Atlas data sets. We further validated this finding in an independent cohort using imaging mass cytometry and found a significant spatial interaction between CAFs and monocytic myeloid cells in the TME. To delineate the interplay between CAFs and monocytic myeloid cells, we used chemotaxis assays to show that LSCC patient–derived CAFs promoted recruitment of CCR2+ monocytes via CCL2, which could be reversed by CCR2 inhibition. Using a three-dimensional culture system, we found that CAFs polarized monocytes to adopt a myeloid-derived suppressor cell (MDSC) phenotype, characterized by robust suppression of autologous CD8+ T-cell proliferation and IFNγ production. We further demonstrated that inhibiting IDO1 and NADPH oxidases, NOX2 and NOX4, restored CD8+ T-cell proliferation by reducing reactive oxygen species (ROS) generation in CAF-induced MDSCs. Taken together, our study highlights a pivotal role of CAFs in regulating monocyte recruitment and differentiation and demonstrated that CCR2 inhibition and ROS scavenging abrogate the CAF–MDSC axis, illuminating a potential therapeutic path to reversing the CAF-mediated immunosuppressive microenvironment.
Data from Systemic Interferon-γ Increases MHC Class I Expression and T-cell Infiltration in Cold Tumors: Results of a Phase 0 Clinical Trial
Authors: Zhang Shihong , Kohli Karan , Black R. Graeme , Yao Lu , Spadinger Sydney M. , He Qianchuan , Pillarisetty Venu G. , Cranmer Lee D. , Van Tine Brian A. , Yee Cassian , Pierce Robert H. , Riddell Stanley R. , Jones Robin L. , Pollack Seth M.
Published: April 2023

Abstract
Interferon-γ (IFNγ) has been studied as a cancer treatment with limited evidence of clinical benefit. However, it could play a role in cancer immunotherapy combination treatments. Despite high expression of immunogenic cancer–testis antigens, synovial sarcoma (SS) and myxoid/round cell liposarcoma (MRCL) have a cold tumor microenvironment (TME), with few infiltrating T cells and low expression of major histocompatibility complex class I (MHC-I). We hypothesized that IFNγ treatment could drive inflammation in a cold TME, facilitating further immunotherapy. We conducted a phase 0 clinical trial treating 8 SS or MRCL patients with weekly systemic IFNγ. We performed pre- and posttreatment biopsies. IFNγ changed the SS and MRCL TME, inducing tumor-surface MHC-I expression and significant T-cell infiltration (P < 0.05). Gene-expression analysis suggested increased tumor antigen presentation and less exhausted phenotypes of the tumor-infiltrating T cells. Newly emergent antigen-specific humoral and/or T-cell responses were found in 3 of 7 evaluable patients. However, increased expression of PD-L1 was observed on tumor-infiltrating myeloid cells and in some cases tumor cells. These findings suggest that systemic IFNγ used to convert SS and MRCL into “hot” tumors will work in concert with anti–PD-1 therapy to provide patient benefit.
Data from Inhibition of MET Signaling with Ficlatuzumab in Combination with Chemotherapy in Refractory AML: Clinical Outcomes and High-Dimensional Analysis
Authors: Wang Victoria E. , Blaser Bradley W. , Patel Ravi K. , Behbehani Gregory K. , Rao Arjun A. , Durbin-Johnson Blythe , Jiang Tommy , Logan Aaron C. , Settles Matthew , Mannis Gabriel N. , Olin Rebecca , Damon Lloyd E. , Martin Thomas G. , Sayre Peter H. , Gaensler Karin M. , McMahon Emma , Flanders Michael , Weinberg Vivian , Ye Chun J. , Carbone David P. , Munster Pamela N. , Fragiadakis Gabriela K. , McCormick Frank , Andreadis Charalambos
Published: April 2023

Abstract
Acute myeloid leukemia (AML) patients refractory to induction therapy or relapsed within 1 year have poor outcomes. Autocrine production of hepatocyte growth factor by myeloid blasts drives leukemogenesis in preclinical models. A phase Ib trial evaluated ficlatuzumab, a first-in-class anti-HGF antibody, in combination with cytarabine in this high-risk population. Dose-limiting toxicities were not observed, and 20 mg/kg was established as the recommended phase II dose. The most frequent treatment-related adverse event was febrile neutropenia. Among 17 evaluable patients, the overall response rate was 53%, all complete remissions. Phospho-proteomic mass cytometry showed potent on-target suppression of p-MET after ficlatuzumab treatment and that attenuation of p-S6 was associated with clinical response. Multiplexed single-cell RNA sequencing using prospectively acquired patient specimens identified IFN response genes as adverse predictive factors. The ficlatuzumab and cytarabine combination is well tolerated, with favorable efficacy. High-dimensional analyses at single-cell resolution represent promising approaches for identifying biomarkers of response and mechanisms of resistance in prospective clinical studies.
Significance:
This study demonstrates a favorable safety profile and promising clinical activity of ficlatuzumab and cytarabine in high-risk AML, thus supporting further investigation of this combination in a randomized trial. It also shows the utility of a novel application using multiplexed single-cell analyses to detect on-target activity and identify biomarkers of response.

This article is highlighted in the In This Issue feature, p. 403
Data from Disease- and Therapy-Specific Impact on Humoral Immune Responses to COVID-19 Vaccination in Hematologic Malignancies
Authors: Chung David J. , Shah Gunjan L. , Devlin Sean M. , Ramanathan Lakshmi V. , Doddi Sital , Pessin Melissa S. , Hoover Elizabeth , Marcello LeeAnn T. , Young Jennifer C. , Boutemine Sawsan R. , Serrano Edith , Sharan Saumya , Momotaj Saddia , Margetich Lauren , Bravo Christina D. , Papanicolaou Genovefa A. , Kamboj Mini , Mato Anthony R. , Roeker Lindsey E. , Hultcrantz Malin , Mailankody Sham , Lesokhin Alexander M. , Vardhana Santosha A. , Knorr David A.
Published: April 2023

Abstract
Coronavirus disease-19 (COVID-19) vaccine response data for patients with hematologic malignancy, who carry high risk for severe COVID-19 illness, are incomplete. In a study of 551 hematologic malignancy patients with leukemia, lymphoma, and multiple myeloma, anti–SARS-CoV-2 spike IgG titers and neutralizing activity were measured at 1 and 3 months from initial vaccination. Compared with healthy controls, patients with hematologic malignancy had attenuated antibody titers at 1 and 3 months. Furthermore, patients with hematologic malignancy had markedly diminished neutralizing capacity of 26.3% at 1 month and 43.6% at 3 months, despite positive seroconversion rates of 51.5% and 68.9% at the respective time points. Healthy controls had 93.2% and 100% neutralizing capacity at 1 and 3 months, respectively. Patients with leukemia, lymphoma, and multiple myeloma on observation had uniformly blunted responses. Treatment with Bruton tyrosine kinase inhibitors, venetoclax, phosphoinositide 3-kinase inhibitors, anti-CD19/CD20–directed therapies, and anti-CD38/B-cell maturation antigen–directed therapies substantially hindered responses, but single-agent immunomodulatory agents did not.
Significance:
Patients with hematologic malignancy have compromised COVID-19 vaccine responses at baseline that are further suppressed by active therapy, with many patients having insufficient neutralizing capacity despite positive antibody titers. Refining vaccine response parameters is critical to guiding clinical care, including the indication for booster vaccines, for this vulnerable population.

See related article by Tamari et al., p. 577.

This article is highlighted in the In This Issue feature, p. 549
Supplementary Figure 3 from Remodeling of the Tumor Microenvironment Through PAK4 Inhibition Sensitizes Tumors to Immune Checkpoint Blockade
Authors: Abril-Rodriguez Gabriel , Torrejon Davis Y. , Karin Daniel , Campbell Katie M. , Medina Egmidio , Saco Justin D. , Galvez Mildred , Champhekar Ameya S. , Perez-Garcilazo Ivan , Baselga-Carretero Ignacio , Singh Jas , Comin-Anduix Begoña , Puig-Saus Cristina , Ribas Antoni
Published: April 2023

Supplementary Figure 3: No difference in signalling through different stimuli between PAK4 KO and WT cells.
Supplementary Figure 2 from Protein Kinase C Alpha is a Central Node for Tumorigenic Transcriptional Networks in Human Prostate Cancer
Authors: Cooke Mariana , Zhang Xuyao , Zhang Suli , Eruslanov Evgeniy , Lal Priti , Daniel Reba E. , Feldman Michael D. , Abba Martin C. , Kazanietz Marcelo G.
Published: April 2023

Cell cycle analysis upon PKCalpha RNAi silencing. Representative Rb phosphorylation as well as cell cycle distribution analysis by FACS in PKCalpha silenced PC3 cells are shown.
Supplementary Data S1 from STAG2 Expression is Associated with Adverse Survival Outcomes and Regulates Cell Phenotype in Muscle-invasive Bladder Cancer
Authors: Athans Sarah R. , Krishnan Nithya , Ramakrishnan Swathi , Cortes Gomez Eduardo , Lage-Vickers Sofía , Rak Monika , Kazmierczak Zara I. , Ohm Joyce Ellen , Attwood Kristopher , Wang Jianmin , Woloszynska Anna
Published: April 2023

Supplementary methods, Supplementary Figures S1-S3, and Supplementary tables
Supplementary Table ST1 from Trends of Ovarian Cancer Incidence by Histotype and Race/Ethnicity in the United States 1992–2019
Authors: Phung Minh Tung , Pearce Celeste Leigh , Meza Rafael , Jeon Jihyoun
Published: April 2023

Supplementary Table 1 shows Registries included in SEER-12 and SEER-17
Supplemental Table 16 from Association Between Recreational Physical Activity and mTOR Signaling Pathway Protein Expression in Breast Tumor Tissue
Authors: Cheng Ting-Yuan David , Zhang Runzhi , Gong Zhihong , Qin Bo , Cannioto Rikki A. , Datta Susmita , Zhang Weizhou , Omilian Angela R. , Yao Song , Khoury Thaer , Hong Chi-Chen , Bandera Elisa V. , Ambrosone Christine B.
Published: April 2023

Supplemental Table 16 reported stratified analysis for high (above the median) and low (at or below the median) total energy intake levels.
Supplementary Figure SF2 from Analysis of N-linked Glycan Alterations in Tissue and Serum Reveals Promising Biomarkers for Intrahepatic Cholangiocarcinoma
Authors: Ochoa-Rios Shaaron , Blaschke Calvin R.K. , Wang Mengjun , Peterson Kendell D. , DelaCourt Andrew , Grauzam Stéphane Elie , Lewin David , Angel Peggi , Roberts Lewis R. , Drake Richard , Mehta Anand S.
Published: April 2023

Patient demographics for A. TMA 1 and B. TMA 2. C. Patient demographics for serum cohort. ALT (Alanine transaminase), AST (aspartate aminotransferase), AFP (Alpha-fetoprotein), and ALP (Alkaline phosphatase). Other liver diseases include nonalcoholic steatohepatitis, hepatitis C with cirrhosis, hepatic adenoma, benign fibrotic gallbladder disease, and diabetes. PSC (Primary Sclerosing cholangitis), HCC (Hepatocellular carcinoma), and OLD (Other liver diseases). Gray shading for missing clinical information. D. Representative N-glycan images of TMA 1 (top) and TMA 2 (bottom) of 2012.717m/z (left) and 1809.646m/z (right). Red boxes select for CCA samples. E. Table details other pathology diagnoses included in TMA 1 with the proposed structure for the N-glycans highly expressed in each. These modifications are characterized based on 1-2 patients.
Supplementary Figure S2 from The Ubiquitin-specific Protease USP36 Associates with the Microprocessor Complex and Regulates miRNA Biogenesis by SUMOylating DGCR8
Authors: Li Yanping , Carey Timothy S. , Feng Catherine H. , Zhu Hong-Ming , Sun Xiao-Xin , Dai Mu-Shui
Published: April 2023

Supplementary Fig. S2 shows that DGCR8 and Drosha can localize in the nucleolus and USP36 can colocalize with DGCR8 in the nucleolus.
Figure S3 from S100A8/S100A9 Promote Progression of Multiple Myeloma via Expansion of Megakaryocytes
Authors: Lin Cindy , Garcia-Gerique Laura , Bonner Erin E. , Mastio Jerome , Rosenwasser Matthew , Cruz Zachary , Lawler Michael , Bernabei Luca , Muthumani Kar , Liu Qin , Poncz Mortimer , Vogl Thomas , Törngren Marie , Eriksson Helena , Vogl Dan T. , Gabrilovich Dmitry I. , Nefedova Yulia
Published: April 2023

Effect of TPO and S100A9 on in vitro MK differentiation.
Figure S1 from S100A8/S100A9 Promote Progression of Multiple Myeloma via Expansion of Megakaryocytes
Authors: Lin Cindy , Garcia-Gerique Laura , Bonner Erin E. , Mastio Jerome , Rosenwasser Matthew , Cruz Zachary , Lawler Michael , Bernabei Luca , Muthumani Kar , Liu Qin , Poncz Mortimer , Vogl Thomas , Törngren Marie , Eriksson Helena , Vogl Dan T. , Gabrilovich Dmitry I. , Nefedova Yulia
Published: April 2023

MK count in the BM of tumor-free mice treated with TQ.
Supplemental Figure 1 from Digital spatial profiling links beta-2-microglobulin expression with immune checkpoint blockade outcomes in head and neck squamous cell carcinoma
Authors: Gavrielatou Niki , Vathiotis Ioannis A. , Aung Thazin Nwe , Shafi Saba , Burela Sneha , Fernandez Aileen I. , Moutafi Myrto , Burtness Barbara A. , Economopoulou Panagiota , Anastasiou Maria , Foukas Periklis , Psyrri Amanda , Rimm David L.
Published: April 2023

signal to noise ratio plots for tumor, leukocyte and macrophage compartments
Supplementary Table S1 from Whole-genome and epigenomic landscapes of malignant gastrointestinal stromal tumors harboring KIT exon 11 557–558 deletion mutations
Authors: Ohshima Keiichi , Nagashima Takeshi , Fujiya Keiichi , Hatakeyama Keiichi , Watanabe Yuko , Morimoto Kimiko , Kamada Fukumi , Shimoda Yuji , Ohnami Sumiko , Naruoka Akane , Serizawa Masakuni , Ohnami Shumpei , Kenmotsu Hirotsugu , Shiomi Akio , Tsubosa Yasuhiro , Bando Etsuro , Sugiura Teiichi , Sugino Takashi , Terashima Masanori , Uesaka Katsuhiko , Urakami Kenichi , Akiyama Yasuto , Yamaguchi Ken
Published: April 2023

Genes for expression signatures.
Data from Phase I Trial of Autologous RNA-electroporated cMET-directed CAR T cells Administered Intravenously in Patients with Melanoma & Breast Carcinoma
Authors: Shah Payal D , Huang Alexander C. , Xu Xiaowei , Orlowski Robert , Amaravadi Ravi K , Schuchter Lynn M. , Zhang Paul , Tchou Julia , Matlawski Tina , Cervini Amanda , Shea Joanne , Gilmore Joan , Lledo Lester , Dengel Karen , Marshall Amy , Wherry E. John , Linette Gerald P , Brennan Andrea Lynne , Gonzalez Vanessa E , Kulikovskaya Irina , Lacey Simon F , Plesa Gabriela , June Carl H , Vonderheide Robert H , Mitchell Tara C
Published: April 2023

Abstract
Purpose: Treatments are limited for metastatic melanoma (MM) and triple-negative breast cancer (mTNBC). This pilot phase I trial (NCT03060356) examined the safety and feasibility of intravenous RNA- electroporated chimeric antigen receptor (CAR) T cells targeting the cell-surface antigen cMET.

Patients and Methods:
MM or mTNBC subjects had at least 30% tumor expression of cMET, measurable disease and progression on prior therapy. Patients received up to 6 infusions (1x10e8 T cells / dose) of CAR T cells without lymphodepleting chemotherapy. Forty-eight percent of prescreened subjects met the cMET expression threshold. Seven (3 MM, 4 mTNBC) were treated.

Results:
Mean age was 50 years (35-64); median ECOG 0 (0-1); median prior lines of chemotherapy/immunotherapy were 4/0 for TNBC and 1/3 for melanoma subjects. Six patients experienced grade (G) 1 or 2 toxicity. Toxicities in at least 1 patient included anemia, fatigue and malaise. One subject had grade 1 cytokine release syndrome. No G 3 or higher toxicity, neurotoxicity, or treatment discontinuation occurred. Best response was stable disease in 4 and disease progression in 3 subjects. Messenger RNA signals corresponding to CAR T cells were detected by RT-PCR in all patients’ blood including in 3 subjects on Day +1 (no infusion administered on this day). Five subjects underwent post-infusion biopsy with no CAR T cell signals seen in tumor. Three subjects had paired tumor tissue; immunohistochemistry showed increases in CD8 and CD3 and decreases in pS6 and Ki67.

Conclusions:
Intravenous administration of RNA-electroporated cMET-directed CAR T cells is safe and feasible.
Data from Efficacy of Low-Dose Aspirin in Colorectal Cancer Risk Prevention is Dependent on ADH1B and ALDH2 Genotype in Japanese Familial Adenomatous Polyposis Patients
Authors: Mure Kanae , Ishikawa Hideki , Mutoh Michihiro , Horinaka Mano , Otani Takahiro , Suzuki Sadao , Wakabayashi Keiji , Sakai Toshiyuki , Sato Yasushi , Doyama Hisashi , Tajika Masahiro , Tanaka Shinji , Horimatsu Takahiro , Takeuchi Yoji , Kashida Hiroshi , Tashiro Jun , Ezoe Yasumasa , Nakajima Takeshi , Ikematsu Hiroaki , Hori Shinichiro , Takayama Tetsuji , Ohda Yoshio
Published: April 2023

Aspirin has gained great attention as a cancer preventive agent. Our previous study revealed that the low-dose aspirin prevents colorectal tumor recurrence in Japanese patients with colorectal adenomas and/or adenocarcinomas, whereas aspirin increases risks in smokers and has no effects on regular drinkers. Our recent study revealed that aspirin reduces polyp growth in Japanese patients with familial adenomatous polyposis (FAP). In this study, we have studied the association of genotypes of alcohol metabolizing enzymes (ADH1B and ALDH2) on aspirin's efficacy of suppressing polyp growth (≥5 mm) in a total of 81 Japanese patients with FAP. Our study revealed that aspirin showed significant preventive effects for patients with ADH1B-AA and AA+GA types [OR = 0.21; 95% confidence interval (CI), 0.05–0.95, and OR = 0.31; 95% CI, 0.10–0.95, respectively], and for patients with ALDH2-GG and GG+GA types (OR = 0.10; 95% CI, 0.01–0.92, and OR = 0.29; 95% CI, 0.09–0.94, respectively), but not for patients with ADH1B-GG and GA+GG types, and ALDH2-AA and GA+AA types. In addition, substantial preventive effects of aspirin were seen for patients with ADH1B-AA type who do not drink regularly (<3 times/week, OR = 0.11; 95% CI, 0.02–0.78), where a statistically significant interaction between aspirin and ADH1B was observed (Pinteraction = 0.036). Results from this exploratory study strongly indicate that aspirin is beneficial in prevention of polyp growth for patients with FAP with ADH1B-AA and AA+GA types, and ALDH2-GG and GG+GA types. Taken together, we propose ADH1B and ALDH2 as candidate markers for the personalized prevention by aspirin.
Significance:
Aspirin is beneficial to patients with FAP with ADH1B-AA and AA+GA types or ALDH2-GG and GG+GA types. ADH1B and ALDH2 genotypes can be the markers for the personalized prevention of colorectal cancer by aspirin.
Data from Chimeric Antigen Receptors Directed at Mutant KRAS Exhibit an Inverse Relationship Between Functional Potency and Neoantigen Selectivity
Authors: Tokatlian Talar , Asuelime Grace E. , Naradikian Martin S. , Mock Jee-Young , Daris Mark E. , Martin Aaron D. , Toledo Warshaviak Dora , Kamb Alexander , Hamburger Agnes E.
Published: April 2023

Neoantigens are among the most intriguing potential immuno-oncology targets because, unlike many cancer targets that are expressed on normal tissues, they are by definition restricted to cancer cells. Medicines directed at common neoantigens such as mutant KRAS are especially interesting because they may offer the convenience and cost of an off-the-shelf therapy. However, all common KRAS mutations produce proteins that differ from the wild type at a single amino acid, creating challenges for molecular discrimination. We have undertaken an effort to optimize single-chain variable fragments (scFv) against peptide/major histocompatibility antigen complexes composed of HLA-A*11 and either G12V- or G12D-mutant KRAS peptides. These scFvs could in principle be used in chimeric antigen receptor (CAR) T-cell therapies for selected patients whose tumors bear either of these mutations. Here we show that optimization of such CARs involves a trade-off between potency and selectivity. We further show that targeting this family without high selectivity engenders risks of cross-reactivity against other members of the G-protein family to which KRAS belongs.
Significance:
We report an effort to generate high potency, selective CARs directed at mutant KRAS peptides. Although the heavily optimized CARs maintain high selectivity against wild-type KRAS, they lose selectivity against other KRAS-related peptides derived from human proteins. To our knowledge, this work is the first to examine the trade-off between potency and selectivity with regard to KRAS pMHC-directed CARs, illustrating the challenge to achieve both sufficient potency and high selectivity.