473 results on '"Freeman GJ"'
Search Results
2. P16-08. Combined blockade of the PD-1 and IL-10 pathways synergistically enhance HIV-specific CD4 T cell functions
- Author
-
Walker BD, Freeman GJ, Kavanagh DG, Pavlik DF, Tighe DP, Kwon DS, Porichis F, and Kaufmann DE
- Subjects
Immunologic diseases. Allergy ,RC581-607 - Published
- 2009
- Full Text
- View/download PDF
3. Early signaling defects in human T cells anergized by T cell presentation of autoantigen.
- Author
-
LaSalle, JM, Tolentino, PJ, Freeman, GJ, Nadler, LM, and Hafler, DA
- Subjects
Antigen-Presenting Cells ,T-Lymphocytes ,Humans ,Calcium ,Tetradecanoylphorbol Acetate ,Receptors ,Antigen ,T-Cell ,Antigens ,Differentiation ,T-Lymphocyte ,HLA-DR Antigens ,Autoantigens ,Cytokines ,Immune Tolerance ,CD3 Complex ,Antigens ,Differentiation ,T-Lymphocyte ,Receptors ,Antigen ,T-Cell ,Medical and Health Sciences ,Immunology - Abstract
Major histocompatibility complex class II-positive human T cell clones are nontraditional antigen-presenting cells (APCs) that are able to simultaneously present and respond to peptide or degraded antigen, but are unable to process intact protein. Although T cell presentation of peptide antigen resulted in a primary proliferative response, T cells that had been previously stimulated by T cells presenting antigen were completely unresponsive to antigen but not to interleukin 2 (IL-2). In contrast, peptide antigen presented by B cells or DR2+ L cell transfectants resulted in T cell activation and responsiveness to restimulation. The anergy induced by T cell presentation of peptide could not be prevented by the addition of either autologous or allogeneic B cells or B7+ DR2+ L cell transfectants, suggesting that the induction of anergy could occur in the presence of costimulation. T cell anergy was induced within 24 h of T cell presentation of antigen and was long lasting. Anergized T cells expressed normal levels of T cell receptor/CD3 but were defective in their ability to release [Ca2+]i to both alpha CD3 and APCs. Moreover, anergized T cells did not proliferate to alpha CD2 monoclonal antibodies or alpha CD3 plus phorbol myristate acetate (PMA), nor did they synthesize IL-2, IL-4, or interferon gamma mRNA in response to either peptide or peptide plus PMA. In contrast, ionomycin plus PMA induced both normal proliferative responses and synthesis of cytokine mRNA, suggesting that the signaling defect in anergized cells occurs before protein kinase C activation and [Ca2+]i release.
- Published
- 1992
4. Expression of lectin-like transcript-1 in human tissues
- Author
-
Llibre, A, Garner, L, Partridge, A, Freeman, GJ, Klenerman, P, and Willberg, CB
- Subjects
Lectin-Like Transcript 1 (LLT1) ,distribution ,C-type lectins ,Articles ,immune-privilege ,human ,natural killer cell ,Immune Response ,Research Article - Abstract
Background: Receptor-ligand pairs of C-type lectin-like proteins have been shown to play an important role in cross talk between lymphocytes, as well as in immune responses within concrete tissues and structures, such as the skin or the germinal centres. The CD161-Lectin-like Transcript 1 (LLT1) pair has gained particular attention in recent years, yet a detailed analysis of LLT1 distribution in human tissue is lacking. One reason for this is the limited availability and poor characterisation of anti-LLT1 antibodies. Methods: We assessed the staining capabilities of a novel anti-LLT1 antibody clone (2H7), both by immunohistochemistry and flow cytometry, showing its efficiency at LLT1 recognition in both settings. We then analysed LLT1 expression in a wide variety of human tissues. Results: We found LLT1 expression in circulating B cells and monocytes, but not in lung and liver-resident macrophages. We found strikingly high LLT1 expression in immune-privileged sites, such as the brain, placenta and testes, and confirmed the ability of LLT1 to inhibit NK cell function. Conclusions: Overall, this study contributes to the development of efficient tools for the study of LLT1. Moreover, its expression in different healthy human tissues and, particularly, in immune-privileged sites, establishes LLT1 as a good candidate as a regulator of immune responses.
- Published
- 2016
5. Ex vivo generation of human anti-pre-B leukemia-specific autologous cytolytic T cells
- Author
-
Cardoso AA, Seamon MJ, Afonso HM, Boussiotis VA, Freeman GJ, Gribben JG, Sallan SE, Nadler LM, GHIA , PAOLO PROSPERO, Cardoso, Aa, Seamon, Mj, Afonso, Hm, Ghia, PAOLO PROSPERO, Boussiotis, Va, Freeman, Gj, Gribben, Jg, Sallan, Se, and Nadler, Lm
- Subjects
Membrane Glycoproteins ,Recombinant Fusion Proteins ,T-Lymphocytes ,Apoptosis ,CHO Cells ,DNA Fragmentation ,Dendritic Cells ,Burkitt Lymphoma ,Immunophenotyping ,Interferon-gamma ,Antigens, CD ,Bone Marrow ,Reference Values ,Cricetinae ,B7-1 Antigen ,Leukemia, B-Cell ,Tumor Cells, Cultured ,Animals ,Humans ,Preleukemia ,B7-2 Antigen ,Lymphocyte Culture Test, Mixed ,Cells, Cultured ,T-Lymphocytes, Cytotoxic - Abstract
In contrast to other neoplasms, antigen-specific autologous cytolytic T cells have not been detected in patients with human pre-B-cell leukemias. The absence of efficient B7 family (B7-1/CD80;B7-2/CD86)-mediated costimulation has been shown to be a major defect in tumor cells' capacity to function as antigen-presenting cells. We show here the generation of autologous anti-pre-B-cell leukemia-specific cytolytic T-cell lines from the marrows of 10 of 15 patients with pre-B-cell malignancies. T-cell costimulation via CD28 is an absolute requirement for the generation of these autologous cytolytic T cells (CTL). Although costimulation could be delivered by either bystander B7 transfectants or professional antigen-presenting cells (indirect costimulation), optimal priming and CTL expansion required that the costimulatory signal was expressed by the tumor cell (direct costimulatory. These anti-pre-B-cell leukemia-specific CTL lysed both unstimulated and CD40-stimulated tumor cells from each patient studied but did not lyse either K562 or CD40-stimulated allogeneic B cells. Cytolysis was mediated by the induction of tumor cell apoptosis by CD8(+) T cells via the perforin-granzyme pathway. Although we were able to generate anti-leukemia-specific CTL from the bone marrow, we were unable to generate such CTL from the peripheral blood of these patients. These studies show that antigen-specific CTL can be generated from the bone marrow of patients with pre-B-cell leukemias and these findings should facilitate the design of adoptive T-cell-mediated immunotherapy trials for the treatment of patients with B-cell precursor malignancies. (C) 1997 by The American Society of Hematology.
- Published
- 1997
6. P16-08. Combined blockade of the PD-1 and IL-10 pathways synergistically enhance HIV-specific CD4 T cell functions
- Author
-
Porichis, F, primary, Kwon, DS, additional, Tighe, DP, additional, Pavlik, DF, additional, Kavanagh, DG, additional, Freeman, GJ, additional, Walker, BD, additional, and Kaufmann, DE, additional
- Published
- 2009
- Full Text
- View/download PDF
7. Pre-B acute lymphoblastic leukemia cells may induce T-cell anergy to alloantigen
- Author
-
Cardoso, AA, primary, Schultze, JL, additional, Boussiotis, VA, additional, Freeman, GJ, additional, Seamon, MJ, additional, Laszlo, S, additional, Billet, A, additional, Sallan, SE, additional, Gribben, JG, additional, and Nadler, LM, additional
- Published
- 1996
- Full Text
- View/download PDF
8. Complete blockade of B7 family-mediated costimulation is necessary to induce human alloantigen-specific anergy: a method to ameliorate graft- versus-host disease and extend the donor pool
- Author
-
Gribben, JG, primary, Guinan, EC, additional, Boussiotis, VA, additional, Ke, XY, additional, Linsley, L, additional, Sieff, C, additional, Gray, GS, additional, Freeman, GJ, additional, and Nadler, LM, additional
- Published
- 1996
- Full Text
- View/download PDF
9. Role of B7-1 in mediating an immune response to myeloid leukemia cells
- Author
-
Matulonis, UA, primary, Dosiou, C, additional, Lamont, C, additional, Freeman, GJ, additional, Mauch, P, additional, Nadler, LM, additional, and Griffin, JD, additional
- Published
- 1995
- Full Text
- View/download PDF
10. CD40 ligand triggered interleukin-6 secretion in multiple myeloma
- Author
-
Urashima, M, primary, Chauhan, D, additional, Uchiyama, H, additional, Freeman, GJ, additional, and Anderson, KC, additional
- Published
- 1995
- Full Text
- View/download PDF
11. Pivotal role of the B7:CD28 pathway in transplantation tolerance and tumor immunity
- Author
-
Guinan, EC, primary, Gribben, JG, additional, Boussiotis, VA, additional, Freeman, GJ, additional, and Nadler, LM, additional
- Published
- 1994
- Full Text
- View/download PDF
12. The B7-2 (B70) costimulatory molecule expressed by monocytes and activated B lymphocytes is the CD86 differentiation antigen
- Author
-
Engel, P, primary, Gribben, JG, additional, Freeman, GJ, additional, Zhou, LJ, additional, Nozawa, Y, additional, Abe, M, additional, Nadler, LM, additional, Wakasa, H, additional, and Tedder, TF, additional
- Published
- 1994
- Full Text
- View/download PDF
13. The gene for B7, a costimulatory signal for T-cell activation, maps to chromosomal region 3q13.3-3q21
- Author
-
Freeman, GJ, primary, Disteche, CM, additional, Gribben, JG, additional, Adler, DA, additional, Freedman, AS, additional, Dougery, J, additional, and Nadler, LM, additional
- Published
- 1992
- Full Text
- View/download PDF
14. Constitutive neuronal expression of the immune regulator, programmed death 1 (PD-1), identified during experimental autoimmune uveitis.
- Author
-
Chen L, Pai V, Levinson R, Sharpe AH, Freeman GJ, Braun J, Gordon LK, Chen, Ling, Pai, Vicky, Levinson, Ralph, Sharpe, Arlene H, Freeman, Gordon J, Braun, Jonathan, and Gordon, Lynn K
- Abstract
Purpose: Programmed death-1 (PD-1) ligation downregulates active lymphocyte responses. The authors tested whether PD-1 or its ligands are expressed in the posterior segment during active intraocular inflammation.Methods: Experimental autoimmune uveitis (EAU) was induced using interphotoreceptor retinoid-binding protein (IRBP 161-180). Ocular inflammation was evaluated by histology and expression of PD-1 ligand tested by immunohistochemistry. PD-1 expression was evaluated by immunohistochemistry, RT-PCR, and Western immunoblotting.Results: Using immunohistochemistry, PD-1, but not its ligands, was constitutively expressed in retinal neurons of naive mouse retina. Both PD-1 and its ligands were observed, as expected, in sites of active inflammation.Conclusions: PD-1 and its ligands were expressed in sites of active inflammation, in accordance with many other models of inflammatory disease. Surprisingly, PD-1, not previously described outside the immune system, was constitutively expressed in retinal neurons, raising the possibility that PD-1 signaling may be important for neuronal function in the absence of an inflammatory insult. [ABSTRACT FROM AUTHOR]- Published
- 2009
- Full Text
- View/download PDF
15. Endothelial programmed death-1 ligand 1 (PD-L1) regulates CD8+ T-cell mediated injury in the heart.
- Author
-
Grabie N, Gotsman I, DaCosta R, Pang H, Stavrakis G, Butte MJ, Keir ME, Freeman GJ, Sharpe AH, Lichtman AH, Grabie, Nir, Gotsman, Israel, DaCosta, Rosa, Pang, Hong, Stavrakis, George, Butte, Manish J, Keir, Mary E, Freeman, Gordon J, Sharpe, Arlene H, and Lichtman, Andrew H
- Published
- 2007
16. Changes in the strength of co-stimulation through the B7/CD28 pathway alter functional T cell responses to altered peptide ligands.
- Author
-
Murtaza, A, Kuchroo, VK, and Freeman, GJ
- Abstract
T cells require a TCR and a co-stimulatory signal for activation. We have examined the effect of the strength of TCR and co-stimulatory signals on proliferation and production of cytokines by differentiated T cell clones. The TCR signal was varied using antigen dose and altered peptide ligands. The co-stimulatory signal was varied by using as antigen-presenting cells, Chinese hamster ovary cell transfectants that express different levels of the B7-1 molecule with similar levels of MHC class II. Our results show that the level of co-stimulation has a profound effect on the response to an antigen, and that a strong co-stimulatory signal can convert a weak agonist into a full agonist and an agonist into a superagonist. Antigenicity is not absolute but a function of the strength of the TCR and co-stimulatory signals. Increasing the strength of co-stimulation can lower antigen concentration required for maximal proliferative responses by T cell clones by 5 log. These results show that the level of expression of co-stimulatory molecules will profoundly regulate T cell clonal expansion and effector functions. [ABSTRACT FROM PUBLISHER]
- Published
- 1999
- Full Text
- View/download PDF
17. Age-associated contraction of tumor-specific T cells impairs antitumor immunity.
- Author
-
Georgiev P, Han S, Huang AY, Nguyen TH, Drijvers JM, Creasey H, Pereira JA, Yao CH, Park JS, Conway TS, Fung ME, Liang D, Peluso M, Joshi S, Rowe JH, Miller BC, Freeman GJ, Sharpe AH, Haigis MC, and Ringel AE
- Abstract
Progressive decline of the adaptive immune system with increasing age coincides with a sharp increase in cancer incidence. In this study, we set out to understand whether deficits in antitumor immunity with advanced age promote tumor progression and/or drive resistance to immunotherapy. We found that multiple syngeneic cancers grew more rapidly in aged versus young adult mice, driven by dysfunctional CD8+ T-cell responses. By systematically mapping immune cell profiles within tumors, we identified loss of tumor antigen-specific CD8+ T cells as a primary feature accelerating the growth of tumors in aged mice and driving resistance to immunotherapy. When antigen-specific T cells from young adult mice were administered to aged mice, tumor outgrowth was delayed and the aged animals became sensitive to PD-1 blockade. These studies reveal how age-associated CD8+ T-cell dysfunction may license tumorigenesis in elderly patients and have important implications for the use of aged mice as pre-clinical models of aging and cancer.
- Published
- 2024
- Full Text
- View/download PDF
18. Targeting METTL3 reprograms the tumor microenvironment to improve cancer immunotherapy.
- Author
-
Yu H, Liu J, Bu X, Ma Z, Yao Y, Li J, Zhang T, Song W, Xiao X, Sun Y, Xiong W, Shi J, Dai P, Xiang B, Duan H, Yan X, Wu F, Zhang WC, Lin D, Hu H, Zhang H, Slack FJ, He HH, Freeman GJ, Wei W, and Zhang J
- Subjects
- Animals, Humans, Mice, Immunotherapy, Methyltransferases genetics, Tumor Microenvironment, Carcinoma, Non-Small-Cell Lung therapy, Lung Neoplasms therapy
- Abstract
The tumor microenvironment (TME) is a heterogeneous ecosystem containing cancer cells, immune cells, stromal cells, cytokines, and chemokines which together govern tumor progression and response to immunotherapies. Methyltransferase-like 3 (METTL3), a core catalytic subunit for RNA N
6 -methyladenosine (m6 A) modification, plays a crucial role in regulating various physiological and pathological processes. Whether and how METTL3 regulates the TME and anti-tumor immunity in non-small-cell lung cancer (NSCLC) remain poorly understood. Here, we report that METTL3 elevates expression of pro-tumorigenic chemokines including CXCL1, CXCL5, and CCL20, and destabilizes PD-L1 mRNA in an m6 A-dependent manner, thereby shaping a non-inflamed TME. Thus, inhibiting METTL3 reprograms a more inflamed TME that renders anti-PD-1 therapy more effective in several murine lung tumor models. Clinically, NSCLC patients who exhibit low-METTL3 expression have a better prognosis when receiving anti-PD-1 therapy. Collectively, our study highlights targeting METTL3 as a promising strategy to improve immunotherapy in NSCLC patients., Competing Interests: Declaration of interests G.J.F. has patents/pending royalties on the PD-1/PD-L1 pathway from Roche, Merck MSD, Bristol-Myers-Squibb, Merck KGA, Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, Eli Lilly, and Novartis. G.J.F. has served on advisory boards for Roche, Bristol-Myers-Squibb, Xios, Origimed, Triursus, iTeos, NextPoint, IgM, Jubilant, Trillium, GV20, IOME, and Geode. G.J.F. has equity in Nextpoint, Triursus, Xios, iTeos, IgM, GV20, Invaria, and Geode. W.W. is a co-founder and consultant for ReKindle Therapeutics. Other authors declare no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)- Published
- 2024
- Full Text
- View/download PDF
19. Affinity fine-tuning anti-CAIX CAR-T cells mitigate on-target off-tumor side effects.
- Author
-
Wang Y, Buck A, Piel B, Zerefa L, Murugan N, Coherd CD, Miklosi AG, Johal H, Bastos RN, Huang K, Ficial M, Laimon YN, Signoretti S, Zhong Z, Hoang SM, Kastrunes GM, Grimaud M, Fayed A, Yuan HC, Nguyen QD, Thai T, Ivanova EV, Paweletz CP, Wu MR, Choueiri TK, Wee JO, Freeman GJ, Barbie DA, and Marasco WA
- Subjects
- Animals, Mice, Humans, Carbonic Anhydrase IX genetics, Antigens, Neoplasm, Antibodies, T-Lymphocytes metabolism, Carcinoma, Renal Cell metabolism, Kidney Neoplasms pathology, Receptors, Chimeric Antigen genetics, Carbonic Anhydrases metabolism, Carbonic Anhydrases therapeutic use
- Abstract
One of the major hurdles that has hindered the success of chimeric antigen receptor (CAR) T cell therapies against solid tumors is on-target off-tumor (OTOT) toxicity due to sharing of the same epitopes on normal tissues. To elevate the safety profile of CAR-T cells, an affinity/avidity fine-tuned CAR was designed enabling CAR-T cell activation only in the presence of a highly expressed tumor associated antigen (TAA) but not when recognizing the same antigen at a physiological level on healthy cells. Using direct stochastic optical reconstruction microscopy (dSTORM) which provides single-molecule resolution, and flow cytometry, we identified high carbonic anhydrase IX (CAIX) density on clear cell renal cell carcinoma (ccRCC) patient samples and low-density expression on healthy bile duct tissues. A Tet-On doxycycline-inducible CAIX expressing cell line was established to mimic various CAIX densities, providing coverage from CAIX-high skrc-59 tumor cells to CAIX-low MMNK-1 cholangiocytes. Assessing the killing of CAR-T cells, we demonstrated that low-affinity/high-avidity fine-tuned G9 CAR-T has a wider therapeutic window compared to high-affinity/high-avidity G250 that was used in the first anti-CAIX CAR-T clinical trial but displayed serious OTOT effects. To assess the therapeutic effect of G9 on patient samples, we generated ccRCC patient derived organotypic tumor spheroid (PDOTS) ex vivo cultures and demonstrated that G9 CAR-T cells exhibited superior efficacy, migration and cytokine release in these miniature tumors. Moreover, in an RCC orthotopic mouse model, G9 CAR-T cells showed enhanced tumor control compared to G250. In summary, G9 has successfully mitigated OTOT side effects and in doing so has made CAIX a druggable immunotherapeutic target., (© 2024. The Author(s).)
- Published
- 2024
- Full Text
- View/download PDF
20. PD-1 Expression on Intratumoral Regulatory T Cells Is Associated with Lack of Benefit from Anti-PD-1 Therapy in Metastatic Clear-Cell Renal Cell Carcinoma Patients.
- Author
-
Denize T, Jegede OA, Matar S, El Ahmar N, West DJ, Walton E, Bagheri AS, Savla V, Nabil Laimon Y, Gupta S, Vemula SV, Braun DA, Burke KP, Catalano PJ, Freeman GJ, Motzer RJ, Atkins MB, McDermott DF, Sharpe AH, Choueiri TK, and Signoretti S
- Subjects
- Humans, Nivolumab therapeutic use, T-Lymphocytes, Regulatory metabolism, Hepatitis A Virus Cellular Receptor 2 metabolism, Everolimus therapeutic use, Programmed Cell Death 1 Receptor metabolism, Carcinoma, Renal Cell pathology
- Abstract
Purpose: Programmed cell death protein 1 (PD-1) expression on CD8+TIM-3-LAG-3- tumor-infiltrating cells predicts positive response to PD-1 blockade in metastatic clear-cell renal cell carcinoma (mccRCC). Because inhibition of PD-1 signaling in regulatory T cells (Treg) augments their immunosuppressive function, we hypothesized that PD-1 expression on tumor-infiltrating Tregs would predict resistance to PD-1 inhibitors., Experimental Design: PD-1+ Tregs were phenotyped using multiparametric immunofluorescence in ccRCC tissues from the CheckMate-025 trial (nivolumab: n = 91; everolimus: n = 90). Expression of CD8, PD-1, TIM-3, and LAG-3 was previously determined (Ficial and colleagues, 2021). Clinical endpoints included progression-free survival (PFS), overall survival (OS), and objective response rate (ORR)., Results: In the nivolumab (but not everolimus) arm, high percentage of PD-1+ Tregs was associated with shorter PFS (3.19 vs. 5.78 months; P = 0.021), shorter OS (18.1 vs. 27.7 months; P = 0.013) and marginally lower ORR (12.5% vs. 31.3%; P = 0.059). An integrated biomarker (PD-1 Treg/CD8 ratio) was developed by calculating the ratio between percentage of PD-1+Tregs (marker of resistance) and percentage of CD8+PD-1+TIM-3-LAG-3- cells (marker of response). In the nivolumab (but not everolimus) arm, patients with high PD-1 Treg/CD8 ratio experienced shorter PFS (3.48 vs. 9.23 months; P < 0.001), shorter OS (18.14 vs. 38.21 months; P < 0.001), and lower ORR (15.69% vs. 40.00%; P = 0.009). Compared with the individual biomarkers, the PD-1 Treg/CD8 ratio showed improved ability to predict outcomes to nivolumab versus everolimus., Conclusions: PD-1 expression on Tregs is associated with resistance to PD-1 blockade in mccRCC, suggesting that targeting Tregs may synergize with PD-1 inhibition. A model that integrates PD-1 expression on Tregs and CD8+TIM-3-LAG-3- cells has higher predictive value., (©2023 American Association for Cancer Research.)
- Published
- 2024
- Full Text
- View/download PDF
21. The B7:CD28 family and friends: Unraveling coinhibitory interactions.
- Author
-
Burke KP, Chaudhri A, Freeman GJ, and Sharpe AH
- Subjects
- Humans, Friends, T-Lymphocytes, CTLA-4 Antigen metabolism, Immunotherapy, B7-1 Antigen metabolism, Immunoglobulins metabolism, Butyrophilins metabolism, Antigens, CD metabolism, CD28 Antigens metabolism, Autoimmune Diseases
- Abstract
Immune responses must be tightly regulated to ensure both optimal protective immunity and tolerance. Costimulatory pathways within the B7:CD28 family provide essential signals for optimal T cell activation and clonal expansion. They provide crucial inhibitory signals that maintain immune homeostasis, control resolution of inflammation, regulate host defense, and promote tolerance to prevent autoimmunity. Tumors and chronic pathogens can exploit these pathways to evade eradication by the immune system. Advances in understanding B7:CD28 pathways have ushered in a new era of immunotherapy with effective drugs to treat cancer, autoimmune diseases, infectious diseases, and transplant rejection. Here, we discuss current understanding of the mechanisms underlying the coinhibitory functions of CTLA-4, PD-1, PD-L1:B7-1 and PD-L2:RGMb interactions and less studied B7 family members, including HHLA2, VISTA, BTNL2, and BTN3A1, as well as their overlapping and unique roles in regulating immune responses, and the therapeutic potential of these insights., Competing Interests: Declaration of interests A.H.S. has patents/pending royalties on the PD-1 pathway from Roche and Novartis. A.H.S. is on advisory boards for Elpiscience, Bicara, Monopteros, Fibrogen, Alixia, IOME, Corner Therapeutics, Bioentre, GlaxoSmithKline, Amgen, and Janssen. She also is on scientific advisory boards for the Massachusetts General Cancer Center, Program in Cellular and Molecular Medicine at Boston Children’s Hospital, the Human Oncology and Pathogenesis Program at Memorial Sloan Kettering Cancer Center, the Johns Hopkins Bloomberg Kimmel Institute for Cancer Immunotherapy, and the Gladstone Institute and is an academic editor for the Journal of Experimental Medicine. A.H.S. has funding from Quark/Iome; AbbVie; Calico; and Taiwan Bio. G.J.F. has patents/pending royalties on the PD-L1/PD-1 pathway from Roche, Merck MSD, Bristol-Myers-Squibb, Merck KGA, Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, Eli Lilly, and Novartis. G.J.F. has patents or patent applications on the use of PD-L1, PD-L2, PD-1, RGMb, HHLA2, KIR3DL3, and BTNL2 in cancer immunotherapy. G.J.F. has served on advisory boards for iTeos, NextPoint, IgM, GV20, IOME, Bioentre, Santa Ana Bio, Simcere of America, and Geode. G.J.F. has equity in Nextpoint, Triursus, Xios, iTeos, IgM, Trillium, Invaria, GV20, Bioentre, and Geode., (Copyright © 2024 Elsevier Inc. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
22. Immune-restoring CAR-T cells display antitumor activity and reverse immunosuppressive TME in a humanized ccRCC mouse model.
- Author
-
Wang Y, Cho JW, Kastrunes G, Buck A, Razimbaud C, Culhane AC, Sun J, Braun DA, Choueiri TK, Wu CJ, Jones K, Nguyen QD, Zhu Z, Wei K, Zhu Q, Signoretti S, Freeman GJ, Hemberg M, and Marasco WA
- Abstract
One of the major barriers that have restricted successful use of chimeric antigen receptor (CAR) T cells in the treatment of solid tumors is an unfavorable tumor microenvironment (TME). We engineered CAR-T cells targeting carbonic anhydrase IX (CAIX) to secrete anti-PD-L1 monoclonal antibody (mAb), termed immune-restoring (IR) CAR G36-PDL1. We tested CAR-T cells in a humanized clear cell renal cell carcinoma (ccRCC) orthotopic mouse model with reconstituted human leukocyte antigen (HLA) partially matched human leukocytes derived from fetal CD34
+ hematopoietic stem cells (HSCs) and bearing human ccRCC skrc-59 cells under the kidney capsule. G36-PDL1 CAR-T cells, haploidentical to the tumor cells, had a potent antitumor effect compared to those without immune-restoring effect. Analysis of the TME revealed that G36-PDL1 CAR-T cells restored active antitumor immunity by promoting tumor-killing cytotoxicity, reducing immunosuppressive cell components such as M2 macrophages and exhausted CD8+ T cells, and enhancing T follicular helper (Tfh)-B cell crosstalk., Competing Interests: W.A.M. has patents in the PD-1/PDL1 field. G.J.F. has patents/pending royalties on the PD-1/PD-L1 pathway from Roche, Merck MSD, Bristol-Myers-Squibb, Merck KGA, Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, Eli Lilly, and Novartis. G.J.F. has served on advisory boards for Roche, Bristol-Myers-Squibb, Xios, Origimed, Triursus, iTeos, NextPoint, IgM, Jubilant, Trillium, IOME, GV20, Invaria, and Geode. G.J.F. has equity in NextPoint, Triursus, Xios, iTeos, IgM, GV20, Invaria, and Geode. M.H. is a co-founder of Neomer Diagnostics where he is also on the advisory board. K.W. received a sponsored-research agreement from Gilead Sciences. K.W. served as a consultant for Gilead Sciences and Horizon Therapeutics. D.A.B. reports nonfinancial support from Bristol Myers Squibb, honoraria from LM Education/Exchange Services, advisory board fees from Exelixis and AVEO, personal fees from Charles River Associates, Schlesinger Associates, Imprint Science, Insight Strategy, Trinity Group, Cancer Expert Now, Adnovate Strategies, MDedge, CancerNetwork, Catenion, OncLive, Cello Health BioConsulting, PWW Consulting, Haymarket Medical Network, Aptitude Health, ASCO Post/Harborside, Targeted Oncology, and AbbVie, and research support from Exelixis and AstraZeneca, outside of the submitted work. C.J.W. holds equity in BioNTech, Inc., and receives research funding from Pharmacyclics. S.S. reports receiving commercial research grants from Bristol-Myers Squibb, AstraZeneca, Exelixis, and Novartis; is a consultant/advisory board member for Merck, AstraZeneca, Bristol-Myers Squibb, CRISPR Therapeutics AG, AACR, and NCI; receives royalties from Biogenex; and mentored several non-US citizens on research projects with potential funding (in part) from non-US sources/Foreign Components. T.K.C. reports institutional and personal, paid and unpaid support for research, advisory boards, consultancy, and honoraria from AstraZeneca, Aravive, Aveo, Bayer, Bristol Myers-Squibb, Calithera, Circle Pharma, Eisai, EMD Serono, Exelixis, GlaxoSmithKline, IQVA, Infinity, Ipsen, Jansen, Kanaph, Lilly, Merck, Nikang, Nuscan, Novartis, Pfizer, Roche, Sanofi/Aventis, Surface Oncology, Takeda, Tempest, Up-To-Date, and CME events (Peerview, OncLive, MJH, and others), outside the submitted work. T.K.C. has institutional patents filed on molecular alterations and immunotherapy response/toxicity, and ctDNA. T.K.C. has equity in Tempest, Pionyr, Osel, and Precede Bio. T.K.C. has served on committees for NCCN, GU Steering Committee, ASCO/ESMO, ACCRU, and KidneyCan. T.K.C. has medical writing and editorial assistance support may have been funded by Communications companies in part. T.K.C. has mentored several non-US citizens on research projects with potential funding (in part) from non-US sources/Foreign Components. The institution (Dana-Farber Cancer Institute) may have received additional independent funding of drug companies or/and royalties potentially involved in research around the subject matter., (© 2024.)- Published
- 2024
- Full Text
- View/download PDF
23. Antigen presentation deficiency, mesenchymal differentiation, and resistance to immunotherapy in the murine syngeneic CT2A tumor model.
- Author
-
Iorgulescu JB, Ruthen N, Ahn R, Panagioti E, Gokhale PC, Neagu M, Speranza MC, Eschle BK, Soroko KM, Piranlioglu R, Datta M, Krishnan S, Yates KB, Baker GJ, Jain RK, Suvà ML, Neuberg D, White FM, Chiocca EA, Freeman GJ, Sharpe AH, Wu CJ, and Reardon DA
- Subjects
- Humans, Animals, Mice, Janus Kinases, Signal Transduction, STAT Transcription Factors, Interferon-gamma, Immunotherapy, Antigen Presentation, Glioblastoma
- Abstract
Background: The GL261 and CT2A syngeneic tumor lines are frequently used as immunocompetent orthotopic mouse models of human glioblastoma (huGBM) but demonstrate distinct differences in their responses to immunotherapy., Methods: To decipher the cell-intrinsic mechanisms that drive immunotherapy resistance in CT2A-luc and to define the aspects of human cancer biology that these lines can best model, we systematically compared their characteristics using whole exome and transcriptome sequencing, and protein analysis through immunohistochemistry, Western blot, flow cytometry, immunopeptidomics, and phosphopeptidomics., Results: The transcriptional profiles of GL261-luc2 and CT2A-luc tumors resembled those of some huGBMs, despite neither line sharing the essential genetic or histologic features of huGBM. Both models exhibited striking hypermutation, with clonal hotspot mutations in RAS genes ( Kras p.G12C in GL261-luc2 and Nras p.Q61L in CT2A-luc). CT2A-luc distinctly displayed mesenchymal differentiation, upregulated angiogenesis, and multiple defects in antigen presentation machinery (e.g. Tap1 p.Y488C and Psmb8 p.A275P mutations) and interferon response pathways (e.g. copy number losses of loci including IFN genes and reduced phosphorylation of JAK/STAT pathway members). The defect in MHC class I expression could be overcome in CT2A-luc by interferon-γ treatment, which may underlie the modest efficacy of some immunotherapy combinations. Additionally, CT2A-luc demonstrated substantial baseline secretion of the CCL-2, CCL-5, and CCL-22 chemokines, which play important roles as myeloid chemoattractants., Conclusion: Although the clinical contexts that can be modeled by GL261 and CT2A for huGBM are limited, CT2A may be an informative model of immunotherapy resistance due to its deficits in antigen presentation machinery and interferon response pathways., Competing Interests: MLS is equity holder, scientific co-founder, and advisory board member of Immunitas Therapeutics. EAC is an advisor to Amacathera, Bionaut Labs, Genenta, Inc., Insightec, Inc., DNAtrix Inc., Seneca Therapeutics, Synthetic Biologics. EAC has equity options in Bionaut Laboratories, DNAtrix, Immunomic Therapeutics, Seneca Therapeutics, Synthetic Biologics, Ternalys Therapeutics. EAC is co-founder and on the Board of Directors of Ternalys Therapeutics. EAC also is a named inventor on patents related to oncolytic HSV1 and noncoding RNAs. MCS has equity options and is a current employee of GSK. MN is a current employee of AbbVie. DN has stock ownership in Madrigal Pharmaceuticals. RJ received consultant fees from Cur, Elpis, Innocoll, SPARC, and SynDevRx,; owns equity in Accurius, Enlight, and SynDevRx; is on the Board of Trustees of Tekla Healthcare Investors, Tekla Life Sciences Investors, Tekla Healthcare Opportunities Fund, and Tekla World Healthcare Fund; and received a research grant from Boehringer Ingelheim. AS currently has funding from Quark, Merck, AbbVie, Moderna and Vertex unrelated to the submitted work. AS serves on advisory boards for Surface Oncology, SQZ Biotechnologies, Selecta, Elpiscience, Monopteros, Bicara, Fibrogen, and Alixis. AS also is on scientific advisory boards for the Massachusetts General Cancer Center, Program in Cellular and Molecular Medicine at Boston Children’s Hospital, the Human Oncology and Pathogenesis Program at Memorial Sloan Kettering Cancer Center, Glaxo Smith Kline and Janssen. AS is an academic editor for the Journal of Experimental Medicine. AS has patents/pending royalties on the PD-1 pathway from Roche and Novartis. GF has patents/pending royalties on the PD-L1/PD-1 pathway from Roche, Merck MSD, Bristol-Myers-Squibb, Merck KGA, Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, Eli Lilly, and Novartis. GF has served on advisory boards for Roche, Bristol-Myers-Squibb, Origimed, Triursus, iTeos, NextPoint, IgM, Jubilant, Trillium, GV20, IOME, and Geode. GF has equity in Nextpoint, Triursus, Xios, iTeos, IgM, Trillium, Invaria, GV20, and Geode. CW is an equity holder of BioNTech, Inc. DR receives institutional support through Dana-Farber Cancer Institute from Acerta Phamaceuticals, Agenus, Bristol-Myers Squibb, Celldex, EMD Serono, Enterome, Epitopoietic Research Corporation, Incyte, Inovio, Insightec, Novartis, Omniox, and Tragara; and is an advisor/consultant for Abbvie, Advantagene, Agenus, Agios, Amgen, AnHeart Therapeutics, Avita Biomedical, Bayer, Boston Biomedical, Boehringer Ingelheim, Bristol-Myers Squibb, Celldex, Deciphera, Del Mar Pharma, DNAtrix, Ellipses Pharma, EMD Serono, Genenta, Genentech/Roche, Hoffman-LaRoche, Imvax, Inovio, Kintara, Kiyatec, Medicenna Biopharma, Merck, Merck KGaA, Monteris, Neuvogen, Novartis, Novocure, Oncorus, Oxigene, Regeneron, Stemline, Sumitono Dainippon Pharma, Pyramid, Taiho Oncology, Vivacitas Oncology, and Y-mabs Therapeutics. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Iorgulescu, Ruthen, Ahn, Panagioti, Gokhale, Neagu, Speranza, Eschle, Soroko, Piranlioglu, Datta, Krishnan, Yates, Baker, Jain, Suvà, Neuberg, White, Chiocca, Freeman, Sharpe, Wu and Reardon.)
- Published
- 2023
- Full Text
- View/download PDF
24. Formate Supplementation Enhances Antitumor CD8+ T-cell Fitness and Efficacy of PD-1 Blockade.
- Author
-
Rowe JH, Elia I, Shahid O, Gaudiano EF, Sifnugel NE, Johnson S, Reynolds AG, Fung ME, Joshi S, LaFleur MW, Park JS, Pauken KE, Rabinowitz JD, Freeman GJ, Haigis MC, and Sharpe AH
- Subjects
- Humans, CD8-Positive T-Lymphocytes metabolism, Formates, Dietary Supplements, Tumor Microenvironment, Programmed Cell Death 1 Receptor metabolism, Neoplasms genetics
- Abstract
The tumor microenvironment (TME) restricts antitumor CD8+ T-cell function and immunotherapy responses. Cancer cells compromise the metabolic fitness of CD8+ T cells within the TME, but the mechanisms are largely unknown. Here we demonstrate that one-carbon (1C) metabolism is enhanced in T cells in an antigen-specific manner. Therapeutic supplementation of 1C metabolism using formate enhances CD8+ T-cell fitness and antitumor efficacy of PD-1 blockade in B16-OVA tumors. Formate supplementation drives transcriptional alterations in CD8+ T-cell metabolism and increases gene signatures for cellular proliferation and activation. Combined formate and anti-PD-1 therapy increases tumor-infiltrating CD8+ T cells, which are essential for enhanced tumor control. Our data demonstrate that formate provides metabolic support to CD8+ T cells reinvigorated by anti-PD-1 to overcome a metabolic vulnerability in 1C metabolism in the TME to further improve T-cell function., Significance: This study identifies that deficiencies in 1C metabolism limit the efficacy of PD-1 blockade in B16-OVA tumors. Supplementing 1C metabolism with formate during anti-PD-1 therapy enhances CD8+ T-cell fitness in the TME and CD8+ T-cell-mediated tumor clearance. These findings demonstrate that formate supplementation can enhance exhausted CD8+ T-cell function. See related commentary by Lin et al., p. 2507. This article is featured in Selected Articles from This Issue, p. 2489., (©2023 American Association for Cancer Research.)
- Published
- 2023
- Full Text
- View/download PDF
25. Regulation of HHLA2 expression in kidney cancer and myeloid cells.
- Author
-
Shigemura T, Perrot N, Huang Z, Bhatt RS, Sheshdeh AB, Ahmar NE, Ghandour F, Signoretti S, McDermott DF, Freeman GJ, and Mahoney KM
- Subjects
- Humans, Animals, Mice, Cytokines metabolism, Myeloid Cells metabolism, Immunoglobulins genetics, Tumor Microenvironment, Carcinoma, Renal Cell genetics, Endogenous Retroviruses metabolism, Kidney Neoplasms genetics
- Abstract
Background: The immune checkpoint HERV-H LTR-associating 2 (HHLA2) is expressed in kidney cancer and various other tumor types. Therapeutics targeting HHLA2 or its inhibitory receptor KIR3DL3 are being developed for solid tumors, including renal cell carcinoma (RCC). However, the regulation of HHLA2 expression remains poorly understood. A better understanding of HHLA2 regulation in tumor cells and the tumor microenvironment is crucial for the successful translation of these therapeutic agents into clinical applications., Methods: Flow cytometry and quantitative real-time PCR were used to analyze HHLA2 expression in primary kidney tumors ex vivo and during in vitro culture. HHLA2 expression in A498 and 786-O ccRCC cell lines was examined in vitro and in subcutaneous tumor xenografts in NSG mice. Monocytes and dendritic cells were analyzed for HHLA2 expression. We tested a range of cytokines and culture conditions, including hypoxia, to induce HHLA2 expression., Results: Analysis of HHLA2 expression revealed that HHLA2 is expressed on tumor cells in primary kidney tumors ex vivo; however, its expression gradually diminishes during a 4-week in vitro culture period. A498 and 786-O ccRCC tumor cell lines do not express HHLA2 in vitro, but HHLA2 expression was observed when grown as subcutaneous xenografts in NSG immunodeficient mice. Induction experiments using various cytokines and culture conditions failed to induce HHLA2 expression in A498 and 786-O tumor cell lines in vitro. Analysis of HHLA2 expression in monocytes and dendritic cells demonstrated that only IL-10 and BMP4, along with IL-1β and IL-6 to a lesser extent, modestly enhanced HHLA2 protein and mRNA expression., Conclusions: HHLA2 expression is induced on kidney cancer cells in vivo by a tumor microenvironmental signal that is not present in vitro. HHLA2 expression is differentially regulated in kidney cancer epithelial cells and monocytes. Cytokines, particularly IL10, that induce HHLA2 expression in monocytes fail to upregulate HHLA2 expression in tumor cell lines in vitro. These findings underscore the importance of the interplay between tumor cell and tumor microenvironmental signals in the regulation of HHLA2. Further investigation is warranted to elucidate the mechanisms involved in HHLA2 regulation and its implications for therapeutic development., (© 2023. The Author(s).)
- Published
- 2023
- Full Text
- View/download PDF
26. The CX3CL1-CX3CR1 chemokine axis can contribute to tumor immune evasion and blockade with a novel CX3CR1 monoclonal antibody enhances response to anti-PD-1 immunotherapy.
- Author
-
Chaudhri A, Bu X, Wang Y, Gomez M, Torchia JA, Hua P, Hung SH, Davies MA, Lizee GA, von Andrian U, Hwu P, and Freeman GJ
- Subjects
- Mice, Animals, Chemokine CX3CL1 metabolism, Antibodies, Monoclonal pharmacology, Antibodies, Monoclonal therapeutic use, Tumor Escape
- Abstract
CX3CL1 secreted in the tumor microenvironment serves as a chemoattractant playing a critical role in metastasis of CX3CR1 expressing cancer cells. CX3CR1 can be expressed in both cancer and immune-inhibitory myeloid cells to facilitate their migration. We generated a novel monoclonal antibody against mouse CX3CR1 that binds to CX3CR1 and blocks the CX3CL1-CX3CR1 interaction. We next explored the immune evasion strategies implemented by the CX3CL1-CX3CR1 axis and find that it initiates a resistance program in cancer cells that results in 1) facilitation of tumor cell migration, 2) secretion of soluble mediators to generate a pro-metastatic niche, 3) secretion of soluble mediators to attract myeloid populations, and 4) generation of tumor-inflammasome. The CX3CR1 monoclonal antibody reduces migration of tumor cells and decreases secretion of immune suppressive soluble mediators by tumor cells. In combination with anti-PD-1 immunotherapy, this CX3CR1 monoclonal antibody enhances survival in an immunocompetent mouse colon carcinoma model through a decrease in tumor-promoting myeloid populations. Thus, this axis is involved in the mechanisms of resistance to anti-PD-1 immunotherapy and the combination therapy can overcome a portion of the resistance mechanisms to anti-PD-1., Competing Interests: GF and AC are inventors of a patent application for the use of anti-CX3CR1 antibody in tumor immunotherapy US20220289854A1. GF has patents/pending royalties on the PD-L1/PD-1 pathway from Roche, Merck MSD, Bristol-Myers-Squibb, Merck KGA, Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, Eli Lilly, and Novartis. GF has served on advisory boards for Roche, Bristol-Myers-Squibb, Origimed, Triursus, iTeos, NextPoint, IgM, Jubilant, Trillium, GV20, IOME, and Geode. GF has equity in Nextpoint, Triursus, Xios, iTeos, IgM, Trillium, Invaria, GV20, and Geode. PHw has served on advisory boards for Dragonfly and Immatics. UvA has received compensation for consulting and SAB membership from AbbVie, Beam, Bluesphere, FL-72, Intergalactic, Moderna, Monopteros, Morphic, Rubius, Selecta Biosciences and SQZ. MD has been a consultant to Roche/Genentech, Array, Pfizer, Novartis, BMS, GSK, Sanofi-Aventis, Vaccinex, Apexigen, Eisai, Iovance, Merck, and ABM Therapeutics. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Chaudhri, Bu, Wang, Gomez, Torchia, Hua, Hung, Davies, Lizee, Andrian, Hwu and Freeman.)
- Published
- 2023
- Full Text
- View/download PDF
27. PD-1 blockade increases the self-renewal of stem-like CD8 T cells to compensate for their accelerated differentiation into effectors.
- Author
-
Gill AL, Wang PH, Lee J, Hudson WH, Ando S, Araki K, Hu Y, Wieland A, Im S, Gavora A, Medina CB, Freeman GJ, Hashimoto M, Reiner SL, and Ahmed R
- Subjects
- Humans, Animals, Mice, Antibodies, Cell Differentiation, Disease Models, Animal, Programmed Cell Death 1 Receptor, CD8-Positive T-Lymphocytes
- Abstract
PD-1
+ TCF-1+ stem-like CD8 T cells act as critical resource cells for maintaining T cell immunity in chronic viral infections and cancer. In addition, they provide the proliferative burst of effector CD8 T cells after programmed death protein 1 (PD-1)-directed immunotherapy. However, it is not known whether checkpoint blockade diminishes the number of these stem-like progenitor cells as effector cell differentiation increases. To investigate this, we used the mouse model of chronic lymphocytic choriomeningitis virus (LCMV) infection. Treatment of chronically infected mice with either αPD-1 or αPD-L1 antibody not only increased effector cell differentiation from the virus-specific stem-like CD8 T cells but also increased their proliferation so their numbers were maintained. The increased self-renewal of LCMV-specific stem-like CD8 T cells was mTOR dependent. We used microscopy to understand the division of these progenitor cells and found that after PD-1 blockade, an individual dividing cell could give rise to a differentiated TCF-1- daughter cell alongside a self-renewing TCF-1+ sister cell. This asymmetric division helped to preserve the number of stem-like cells. Moreover, we found that the PD-1+ TCF-1+ stem-like CD8 T cells retained their transcriptional program and their in vivo functionality in terms of responding to viral infection and to repeat PD-1 blockade. Together, our results demonstrate that PD-1 blockade does not deplete the stem-like population despite increasing effector differentiation. These findings have implications for PD-1-directed immunotherapy in humans.- Published
- 2023
- Full Text
- View/download PDF
28. Inverse relationship between Fusobacterium nucleatum amount and tumor CD274 (PD-L1) expression in colorectal carcinoma.
- Author
-
Ugai T, Shimizu T, Kawamura H, Ugai S, Takashima Y, Usui G, Väyrynen JP, Okadome K, Haruki K, Akimoto N, Masugi Y, da Silva A, Mima K, Zhang X, Chan AT, Wang M, Garrett WS, Freeman GJ, Meyerhardt JA, Nowak JA, Song M, Giannakis M, and Ogino S
- Abstract
Objectives: The CD274 (programmed cell death 1 ligand 1, PD-L1)/PDCD1 (programmed cell death 1, PD-1) immune checkpoint axis is known to regulate the antitumor immune response. Evidence also supports an immunosuppressive effect of Fusobacterium nucleatum . We hypothesised that tumor CD274 overexpression might be inversely associated with abundance of F. nucleatum in colorectal carcinoma., Methods: We assessed tumor CD274 expression by immunohistochemistry and F. nucleatum DNA within tumor tissue by quantitative PCR in 812 cases among 4465 incident rectal and colon cancer cases that had occurred in two prospective cohort studies. Multivariable logistic regression analyses with inverse probability weighting were used to adjust for selection bias because of tissue data availability and potential confounders including microsatellite instability status, CpG island methylator phenotype, LINE-1 methylation level and KRAS , BRAF and PIK3CA mutations., Results: Fusobacterium nucleatum DNA was detected in tumor tissue in 109 (13%) cases. Tumor CD274 expression level was inversely associated with the amount of F. nucleatum in colorectal cancer tissue ( P = 0.0077). For one category-unit increase in three ordinal F. nucleatum categories (negative vs. low vs. high), multivariable-adjusted odds ratios (with 95% confidence interval) of the low, intermediate and high CD274 categories (vs. negative) were 0.78 (0.41-1.51), 0.64 (0.32-1.28) and 0.50 (0.25-0.99), respectively ( P
trend = 0.032)., Conclusions: Tumor CD274 expression level was inversely associated with the amount of F. nucleatum in colorectal cancer tissue, suggesting that different immunosuppressive mechanisms (i.e. PDCD1 immune checkpoint activation and tumor F. nucleatum enrichment) tend to be used by different tumor subgroups., Competing Interests: ATC previously served as a consultant for Bayer Healthcare and Pfizer Inc. MG was on an advisory board for AstraZeneca and receives research funding from Bristol‐Myers Squibb. JAM has received institutional research funding from Boston Biomedical, has served as an advisor/consultant to Ignyta and COTA Healthcare, and served on a grant review panel for the National Comprehensive Cancer Network funded by Taiho Pharmaceutical. This study was not funded by any of these commercial entities., (© 2023 The Authors. Clinical & Translational Immunology published by John Wiley & Sons Australia, Ltd on behalf of Australian and New Zealand Society for Immunology, Inc.)- Published
- 2023
- Full Text
- View/download PDF
29. Publisher Correction: Targeting PD-L2-RGMb overcomes microbiome-related immunotherapy resistance.
- Author
-
Park JS, Gazzaniga FS, Wu M, Luthens AK, Gillis J, Zheng W, LaFleur MW, Johnson SB, Morad G, Park EM, Zhou Y, Watowich SS, Wargo JA, Freeman GJ, Kasper DL, and Sharpe AH
- Published
- 2023
- Full Text
- View/download PDF
30. Novel Antimurine Thyroid-Stimulating Hormone Receptor Monoclonal Antibodies.
- Author
-
Klee AN, Torchia JA, and Freeman GJ
- Subjects
- Mice, Rats, Animals, Antibodies, Monoclonal, Receptors, Thyrotropin metabolism, Autoantibodies, Thyrotropin, Graves Disease drug therapy, Graves Disease etiology, Hashimoto Disease complications
- Abstract
Autoantibodies against thyroid proteins are present in several thyroid diseases. Thyroid-stimulating hormone receptor (TSHR) is a G-protein-coupled receptor (GPCR) that binds to thyroid-stimulating hormone (TSH) and stimulates production of thyroxine (T4) and triiodothyronine (T3). When agonized by anti-TSHR autoantibodies, aberrant production of thyroid hormone can lead to Graves' Disease (GD). In Hashimoto's thyroiditis (HT), anti-TSHR autoantibodies target the thyroid for immune attack. To better understand the role of anti-TSHR antibodies in thyroid disease, we generated a set of rat antimouse (m)TSHR monoclonal antibodies with a range of affinities, blocking of TSH, and agonist activity. These antibodies could be used to investigate the etiology and therapy of thyroid disease in mouse models and as building blocks in protein therapeutics that target the thyroid for treatment in either HT or GD.
- Published
- 2023
- Full Text
- View/download PDF
31. ERK and USP5 govern PD-1 homeostasis via deubiquitination to modulate tumor immunotherapy.
- Author
-
Xiao X, Shi J, He C, Bu X, Sun Y, Gao M, Xiang B, Xiong W, Dai P, Mao Q, Xing X, Yao Y, Yu H, Xu G, Li S, Ren Y, Chen B, Jiang C, Meng G, Lee YR, Wei W, Freeman GJ, Xie C, and Zhang J
- Subjects
- Animals, Mice, Ubiquitin-Specific Proteases genetics, Ubiquitin-Specific Proteases metabolism, Homeostasis, Immunotherapy, Extracellular Signal-Regulated MAP Kinases, Programmed Cell Death 1 Receptor
- Abstract
The programmed cell death protein 1 (PD-1) is an inhibitory receptor on T cells and plays an important role in promoting cancer immune evasion. While ubiquitin E3 ligases regulating PD-1 stability have been reported, deubiquitinases governing PD-1 homeostasis to modulate tumor immunotherapy remain unknown. Here, we identify the ubiquitin-specific protease 5 (USP5) as a bona fide deubiquitinase for PD-1. Mechanistically, USP5 interacts with PD-1, leading to deubiquitination and stabilization of PD-1. Moreover, extracellular signal-regulated kinase (ERK) phosphorylates PD-1 at Thr234 and promotes PD-1 interaction with USP5. Conditional knockout of Usp5 in T cells increases the production of effector cytokines and retards tumor growth in mice. USP5 inhibition in combination with Trametinib or anti-CTLA-4 has an additive effect on suppressing tumor growth in mice. Together, this study describes a molecular mechanism of ERK/USP5-mediated regulation of PD-1 and identifies potential combinatorial therapeutic strategies for enhancing anti-tumor efficacy., (© 2023. The Author(s).)
- Published
- 2023
- Full Text
- View/download PDF
32. PRMT1 mediated methylation of cGAS suppresses anti-tumor immunity.
- Author
-
Liu J, Bu X, Chu C, Dai X, Asara JM, Sicinski P, Freeman GJ, and Wei W
- Subjects
- Methylation, Nucleotidyltransferases metabolism, Signal Transduction genetics, Methyltransferases metabolism, B7-H1 Antigen genetics, B7-H1 Antigen metabolism, Immunity, Innate genetics
- Abstract
Activation of the cGAS/STING innate immunity pathway is essential and effective for anti-tumor immunotherapy. However, it remains largely elusive how tumor-intrinsic cGAS signaling is suppressed to facilitate tumorigenesis by escaping immune surveillance. Here, we report that the protein arginine methyltransferase, PRMT1, methylates cGAS at the conserved Arg133 residue, which prevents cGAS dimerization and suppresses the cGAS/STING signaling in cancer cells. Notably, genetic or pharmaceutical ablation of PRMT1 leads to activation of cGAS/STING-dependent DNA sensing signaling, and robustly elevates the transcription of type I and II interferon response genes. As such, PRMT1 inhibition elevates tumor-infiltrating lymphocytes in a cGAS-dependent manner, and promotes tumoral PD-L1 expression. Thus, combination therapy of PRMT1 inhibitor with anti-PD-1 antibody augments the anti-tumor therapeutic efficacy in vivo. Our study therefore defines the PRMT1/cGAS/PD-L1 regulatory axis as a critical factor in determining immune surveillance efficacy, which serves as a promising therapeutic target for boosting tumor immunity., (© 2023. The Author(s).)
- Published
- 2023
- Full Text
- View/download PDF
33. Targeting PD-L2-RGMb overcomes microbiome-related immunotherapy resistance.
- Author
-
Park JS, Gazzaniga FS, Wu M, Luthens AK, Gillis J, Zheng W, LaFleur MW, Johnson SB, Morad G, Park EM, Zhou Y, Watowich SS, Wargo JA, Freeman GJ, Kasper DL, and Sharpe AH
- Subjects
- Animals, Humans, Mice, Cell Adhesion Molecules, Neuronal, Disease Models, Animal, Down-Regulation, Fecal Microbiota Transplantation, Germ-Free Life, Immune Checkpoint Inhibitors pharmacology, Immune Checkpoint Inhibitors therapeutic use, Protein Binding drug effects, T-Lymphocytes drug effects, T-Lymphocytes immunology, Drug Resistance, Neoplasm drug effects, Immunotherapy, Melanoma immunology, Melanoma microbiology, Melanoma therapy, Microbiota
- Abstract
The gut microbiota is a crucial regulator of anti-tumour immunity during immune checkpoint inhibitor therapy. Several bacteria that promote an anti-tumour response to immune checkpoint inhibitors have been identified in mice
1-6 . Moreover, transplantation of faecal specimens from responders can improve the efficacy of anti-PD-1 therapy in patients with melanoma7,8 . However, the increased efficacy from faecal transplants is variable and how gut bacteria promote anti-tumour immunity remains unclear. Here we show that the gut microbiome downregulates PD-L2 expression and its binding partner repulsive guidance molecule b (RGMb) to promote anti-tumour immunity and identify bacterial species that mediate this effect. PD-L1 and PD-L2 share PD-1 as a binding partner, but PD-L2 can also bind RGMb. We demonstrate that blockade of PD-L2-RGMb interactions can overcome microbiome-dependent resistance to PD-1 pathway inhibitors. Antibody-mediated blockade of the PD-L2-RGMb pathway or conditional deletion of RGMb in T cells combined with an anti-PD-1 or anti-PD-L1 antibody promotes anti-tumour responses in multiple mouse tumour models that do not respond to anti-PD-1 or anti-PD-L1 alone (germ-free mice, antibiotic-treated mice and even mice colonized with stool samples from a patient who did not respond to treatment). These studies identify downregulation of the PD-L2-RGMb pathway as a specific mechanism by which the gut microbiota can promote responses to PD-1 checkpoint blockade. The results also define a potentially effective immunological strategy for treating patients who do not respond to PD-1 cancer immunotherapy., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2023
- Full Text
- View/download PDF
34. PI3Kβ controls immune evasion in PTEN-deficient breast tumours.
- Author
-
Bergholz JS, Wang Q, Wang Q, Ramseier M, Prakadan S, Wang W, Fang R, Kabraji S, Zhou Q, Gray GK, Abell-Hart K, Xie S, Guo X, Gu H, Von T, Jiang T, Tang S, Freeman GJ, Kim HJ, Shalek AK, Roberts TM, and Zhao JJ
- Subjects
- Animals, Mice, Immunotherapy, Phosphoinositide-3 Kinase Inhibitors, Signal Transduction, Mammary Neoplasms, Experimental enzymology, Mammary Neoplasms, Experimental genetics, Mammary Neoplasms, Experimental immunology, Immune Evasion, Phosphatidylinositol 3-Kinase metabolism, PTEN Phosphohydrolase deficiency, PTEN Phosphohydrolase genetics, Mammary Neoplasms, Animal enzymology, Mammary Neoplasms, Animal genetics, Mammary Neoplasms, Animal immunology
- Abstract
Loss of the PTEN tumour suppressor is one of the most common oncogenic drivers across all cancer types
1 . PTEN is the major negative regulator of PI3K signalling. The PI3Kβ isoform has been shown to play an important role in PTEN-deficient tumours, but the mechanisms underlying the importance of PI3Kβ activity remain elusive. Here, using a syngeneic genetically engineered mouse model of invasive breast cancer driven by ablation of both Pten and Trp53 (which encodes p53), we show that genetic inactivation of PI3Kβ led to a robust anti-tumour immune response that abrogated tumour growth in syngeneic immunocompetent mice, but not in immunodeficient mice. Mechanistically, PI3Kβ inactivation in the PTEN-null setting led to reduced STAT3 signalling and increased the expression of immune stimulatory molecules, thereby promoting anti-tumour immune responses. Pharmacological PI3Kβ inhibition also elicited anti-tumour immunity and synergized with immunotherapy to inhibit tumour growth. Mice with complete responses to the combined treatment displayed immune memory and rejected tumours upon re-challenge. Our findings demonstrate a molecular mechanism linking PTEN loss and STAT3 activation in cancer and suggest that PI3Kβ controls immune escape in PTEN-null tumours, providing a rationale for combining PI3Kβ inhibitors with immunotherapy for the treatment of PTEN-deficient breast cancer., (© 2023. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2023
- Full Text
- View/download PDF
35. mTORC1 upregulates B7-H3/CD276 to inhibit antitumor T cells and drive tumor immune evasion.
- Author
-
Liu HJ, Du H, Khabibullin D, Zarei M, Wei K, Freeman GJ, Kwiatkowski DJ, and Henske EP
- Subjects
- Humans, Genes, Regulator, Transcription Factors, Mechanistic Target of Rapamycin Complex 1, B7 Antigens, Tumor Escape, T-Lymphocytes
- Abstract
Identifying the mechanisms underlying the regulation of immune checkpoint molecules and the therapeutic impact of targeting them in cancer is critical. Here we show that high expression of the immune checkpoint B7-H3 (CD276) and high mTORC1 activity correlate with immunosuppressive phenotypes and worse clinical outcomes in 11,060 TCGA human tumors. We find that mTORC1 upregulates B7-H3 expression via direct phosphorylation of the transcription factor YY2 by p70 S6 kinase. Inhibition of B7-H3 suppresses mTORC1-hyperactive tumor growth via an immune-mediated mechanism involving increased T-cell activity and IFN-γ responses coupled with increased tumor cell expression of MHC-II. CITE-seq reveals strikingly increased cytotoxic CD38
+ CD39+ CD4+ T cells in B7-H3-deficient tumors. In pan-human cancers, a high cytotoxic CD38+ CD39+ CD4+ T-cell gene signature correlates with better clinical prognosis. These results show that mTORC1-hyperactivity, present in many human tumors including tuberous sclerosis complex (TSC) and lymphangioleiomyomatosis (LAM), drives B7-H3 expression leading to suppression of cytotoxic CD4+ T cells., (© 2023. The Author(s).)- Published
- 2023
- Full Text
- View/download PDF
36. Losartan controls immune checkpoint blocker-induced edema and improves survival in glioblastoma mouse models.
- Author
-
Datta M, Chatterjee S, Perez EM, Gritsch S, Roberge S, Duquette M, Chen IX, Naxerova K, Kumar AS, Ghosh M, Emblem KE, Ng MR, Ho WW, Kumar P, Krishnan S, Dong X, Speranza MC, Neagu MR, Iorgulescu JB, Huang RY, Youssef G, Reardon DA, Sharpe AH, Freeman GJ, Suvà ML, Xu L, and Jain RK
- Subjects
- Animals, Mice, Losartan pharmacology, Losartan therapeutic use, Immune Checkpoint Inhibitors adverse effects, CD8-Positive T-Lymphocytes, Edema, Tumor Microenvironment, Glioblastoma pathology
- Abstract
Immune checkpoint blockers (ICBs) have failed in all phase III glioblastoma trials. Here, we found that ICBs induce cerebral edema in some patients and mice with glioblastoma. Through single-cell RNA sequencing, intravital imaging, and CD8
+ T cell blocking studies in mice, we demonstrated that this edema results from an inflammatory response following antiprogrammed death 1 (PD1) antibody treatment that disrupts the blood-tumor barrier. Used in lieu of immunosuppressive corticosteroids, the angiotensin receptor blocker losartan prevented this ICB-induced edema and reprogrammed the tumor microenvironment, curing 20% of mice which increased to 40% in combination with standard of care treatment. Using a bihemispheric tumor model, we identified a "hot" tumor immune signature prior to losartan+anti-PD1 therapy that predicted long-term survival. Our findings provide the rationale and associated biomarkers to test losartan with ICBs in glioblastoma patients.- Published
- 2023
- Full Text
- View/download PDF
37. mTOR regulates T cell exhaustion and PD-1-targeted immunotherapy response during chronic viral infection.
- Author
-
Ando S, Perkins CM, Sajiki Y, Chastain C, Valanparambil RM, Wieland A, Hudson WH, Hashimoto M, Ramalingam SS, Freeman GJ, Ahmed R, and Araki K
- Subjects
- CD8-Positive T-Lymphocytes metabolism, Hepatitis A Virus Cellular Receptor 2 genetics, Hepatitis A Virus Cellular Receptor 2 metabolism, Immunotherapy, Persistent Infection, T-Cell Exhaustion, TOR Serine-Threonine Kinases genetics, TOR Serine-Threonine Kinases metabolism, Programmed Cell Death 1 Receptor genetics, Programmed Cell Death 1 Receptor metabolism, Virus Diseases metabolism
- Abstract
T cell exhaustion is a state of T cell dysfunction associated with expression of programmed death 1 (PD-1). Exhausted CD8+ T cells are maintained by self-renewing stem-like T cells that provide differentiated TIM3+ cells, a part of which possesses effector-like properties. PD-1-targeted therapies enhance T cell response by promoting differentiation of stem-like T cells toward TIM3+ cells, but the role of mTOR during T cell exhaustion remains elusive. Here, we showed that mTOR inhibition has distinct outcomes during the beginning of and after the establishment of chronic viral infection. Blocking mTOR during the T cell expansion phase enhanced the T cell response by causing accumulation of stem-like T cells, leading to improved efficacy of PD-1 immunotherapy; whereas, after exhaustion progressed, mTOR inhibition caused immunosuppression, characterized by decreased TIM3+ cells and increased viral load with minimal changes in stem-like T cells. Mechanistically, a cell-intrinsic mTOR signal was vital for differentiation of stem-like T cells into the TIM3+ state in the early and late phases of chronic infection as well as during PD-1 immunotherapy. Thus, PD-1 blockade worked after cessation of mTOR inhibition, but simultaneous treatment failed to induce functional TIM3+ cells, reducing efficacy of PD-1 immunotherapy. Our data demonstrate that mTOR regulates T cell exhaustion and have important implications for combination cancer therapies with PD-1 blockade.
- Published
- 2023
- Full Text
- View/download PDF
38. STING agonism overcomes STAT3-mediated immunosuppression and adaptive resistance to PARP inhibition in ovarian cancer.
- Author
-
Ding L, Wang Q, Martincuks A, Kearns MJ, Jiang T, Lin Z, Cheng X, Qian C, Xie S, Kim HJ, Launonen IM, Färkkilä A, Roberts TM, Freeman GJ, Liu JF, Konstantinopoulos PA, Matulonis U, Yu H, and Zhao JJ
- Subjects
- Animals, Female, Humans, Mice, Cell Line, Tumor, Immunosuppression Therapy, STAT3 Transcription Factor metabolism, Tumor Microenvironment, Ovarian Neoplasms genetics, Poly(ADP-ribose) Polymerase Inhibitors pharmacology
- Abstract
Background: Poly (ADP-ribose) polymerase (PARP) inhibition (PARPi) has demonstrated potent therapeutic efficacy in patients with BRCA-mutant ovarian cancer. However, acquired resistance to PARPi remains a major challenge in the clinic., Methods: PARPi-resistant ovarian cancer mouse models were generated by long-term treatment of olaparib in syngeneic Brca1-deficient ovarian tumors. Signal transducer and activator of transcription 3 (STAT3)-mediated immunosuppression was investigated in vitro by co-culture experiments and in vivo by analysis of immune cells in the tumor microenvironment (TME) of human and mouse PARPi-resistant tumors. Whole genome transcriptome analysis was performed to assess the antitumor immunomodulatory effect of STING (stimulator of interferon genes) agonists on myeloid cells in the TME of PARPi-resistant ovarian tumors. A STING agonist was used to overcome STAT3-mediated immunosuppression and acquired PARPi resistance in syngeneic and patient-derived xenografts models of ovarian cancer., Results: In this study, we uncover an adaptive resistance mechanism to PARP inhibition mediated by tumor-associated macrophages (TAMs) in the TME. Markedly increased populations of protumor macrophages are found in BRCA-deficient ovarian tumors that rendered resistance to PARPi in both murine models and patients. Mechanistically, PARP inhibition elevates the STAT3 signaling pathway in tumor cells, which in turn promotes protumor polarization of TAMs. STAT3 ablation in tumor cells mitigates polarization of protumor macrophages and increases tumor-infiltrating T cells on PARP inhibition. These findings are corroborated in patient-derived, PARPi-resistant BRCA1-mutant ovarian tumors. Importantly, STING agonists reshape the immunosuppressive TME by reprogramming myeloid cells and overcome the TME-dependent adaptive resistance to PARPi in ovarian cancer. This effect is further enhanced by addition of the programmed cell death protein-1 blockade., Conclusions: We elucidate an adaptive immunosuppression mechanism rendering resistance to PARPi in BRCA1-mutant ovarian tumors. This is mediated by enrichment of protumor TAMs propelled by PARPi-induced STAT3 activation in tumor cells. We also provide a new strategy to reshape the immunosuppressive TME with STING agonists and overcome PARPi resistance in ovarian cancer., Competing Interests: Competing interests: LD, QW and JJZ are co-inventors of the patent related to this study (US application 63/148426). QW is a scientific consultant for Crimson Biopharm. PAK is a scientific consultant for Alkermes, AstraZeneca, Bayer, GSK, Merck, Pfizer, Tesaro, Mersana, Repare, Kadmon and served as the principal investigator (PI) of institutional support clinical trial for AstraZeneca, Bayer, GSK, Merck, Lilly, and BMS. TMR is cofounder of Crimson Biopharm and Geode Therapeutics, and a SAB member for Shiftbio and K2B Therapeutics. GJF has patents/pending royalties on the PD-L1/PD-1 pathway from Roche, Merck MSD, Bristol Myers Squibb, Merck KGA, Boehringer Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, Eli Lilly, and Novartis. GJF has served on advisory boards for Roche, Bristol Myers Squibb, Origimed, Triursus, iTeos, NextPoint, IgM, Jubilant, Trillium, GV20, IOME, and Geode. GJF has equity in Nextpoint, Triursus, Xios, iTeos, IgM, Trillium, Invaria, GV20, and Geode. JFL has served as a scientific consultant for Genentech/Roche and Bristol Myers Squibb, and has been on advisory boards for Clovis Oncology, Genentech/Roche, GlaxoSmithKline, Regeneron, AstraZeneca, and Eisai. JFL has served as the institutional PI (support to the institution) of clinical trials by Genentech/Roche, AstraZeneca, Boston Biomedical, Acetylon Pharmaceuticals, Bristol Myers Squibb, Agenus, CytomX Therapeutics, Regeneron, Tesaro/GSK, Clovis Oncology, Surface Oncology, 2X Oncology, Vigeo Therapeutics, Aravive, Arch Oncology and Zentalis Pharmaceuticals. UM has served on the advisory boards of 2X Oncology, Fujifilm, Immunogen, Mersana, Geneos, and Merck. JJZ is cofounder and director of Crimson Biopharm and Geode Therapeutics. The remaining authors declare no competing interests., (© Author(s) (or their employer(s)) 2023. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)
- Published
- 2023
- Full Text
- View/download PDF
39. Optimized ACE2 decoys neutralize antibody-resistant SARS-CoV-2 variants through functional receptor mimicry and treat infection in vivo.
- Author
-
Torchia JA, Tavares AH, Carstensen LS, Chen DY, Huang J, Xiao T, Mukherjee S, Reeves PM, Tu H, Sluder AE, Chen B, Kotton DN, Bowen RA, Saeed M, Poznansky MC, and Freeman GJ
- Subjects
- Humans, SARS-CoV-2, COVID-19
- Abstract
As severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) evolves to escape natural antibodies, it also loses sensitivity to therapeutic antibody drugs. By contrast, evolution selects for binding to ACE2, the cell-surface receptor required for SARS-CoV-2 infection. Consistent with this, we find that an ACE2 decoy neutralizes antibody-resistant variants, including Omicron, with no loss in potency. To identify design features necessary for in vivo activity, we compare several enzymatically inactive, Fc effector-silenced ACE2-Fc decoys. Inclusion of the ACE2 collectrin-like domain not only improves affinity for the S protein but also unexpectedly extends serum half-life and is necessary to reduce disease severity and viral titer in Syrian hamsters. Fc effector function is not required. The activity of ACE2 decoy receptors is due, in part, to their ability to trigger an irreversible structural change in the viral S protein. Our studies provide a new understanding of how ACE2 decoys function and support their development as therapeutics to treat ACE2-dependent coronaviruses.
- Published
- 2022
- Full Text
- View/download PDF
40. Enterohepatic Shunt-Driven Cholemia Predisposes to Liver Cancer.
- Author
-
Yeoh BS, Saha P, Golonka RM, Zou J, Petrick JL, Abokor AA, Xiao X, Bovilla VR, Bretin ACA, Rivera-Esteban J, Parisi D, Florio AA, Weinstein SJ, Albanes D, Freeman GJ, Gohara AF, Ciudin A, Pericàs JM, Joe B, Schwabe RF, McGlynn KA, Gewirtz AT, and Vijay-Kumar M
- Subjects
- Humans, Mice, Animals, Mice, Inbred C57BL, Prostheses and Implants, Dietary Fiber, Liver Neoplasms etiology, Carcinoma, Hepatocellular etiology, Digestive System Diseases
- Abstract
Background & Aims: Pathogenesis of hepatocellular carcinoma (HCC), which kills millions annually, is poorly understood. Identification of risk factors and modifiable determinants and mechanistic understanding of how they impact HCC are urgently needed., Methods: We sought early prognostic indicators of HCC in C57BL/6 mice, which we found were prone to developing this disease when fed a fermentable fiber-enriched diet. Such markers were used to phenotype and interrogate stages of HCC development. Their human relevance was tested using serum collected prospectively from an HCC/case-control cohort., Results: HCC proneness in mice was dictated by the presence of congenitally present portosystemic shunt (PSS), which resulted in markedly elevated serum bile acids (BAs). Approximately 10% of mice from various sources exhibited PSS/cholemia, but lacked an overt phenotype when fed standard chow. However, PSS/cholemic mice fed compositionally defined diets, developed BA- and cyclooxygenase-dependent liver injury, which was exacerbated and uniformly progressed to HCC when diets were enriched with the fermentable fiber inulin. Such progression to cholestatic HCC associated with exacerbated cholemia and an immunosuppressive milieu, both of which were required in that HCC was prevented by impeding BA biosynthesis or neutralizing interleukin-10 or programmed death protein 1. Analysis of human sera revealed that elevated BA was associated with future development of HCC., Conclusions: PSS is relatively common in C57BL/6 mice and causes silent cholemia, which predisposes to liver injury and HCC, particularly when fed a fermentable fiber-enriched diet. Incidence of silent PSS/cholemia in humans awaits investigation. Regardless, measuring serum BA may aid HCC risk assessment, potentially alerting select individuals to consider dietary or BA interventions., (Copyright © 2022 AGA Institute. All rights reserved.)
- Published
- 2022
- Full Text
- View/download PDF
41. Immunoregulatory effects of RGMb in gut inflammation.
- Author
-
Pérez-Cruz M, Iliopoulou BP, Hsu K, Wu HH, Erkers T, Swaminathan K, Tang SW, Bader CS, Kambham N, Xie B, Dekruyff RH, Freeman GJ, and Meyer E
- Subjects
- Humans, Mice, Animals, Inflammation, Immunosuppressive Agents, Antibodies, Monoclonal pharmacology, Antibodies, Monoclonal therapeutic use, Cell Adhesion Molecules, Neuronal, Graft vs Host Disease, Inflammatory Bowel Diseases therapy, Colitis chemically induced
- Abstract
Graft-versus-host disease (GvHD) is a major complication after allogeneic hematopoietic cell transplantation (HCT). Current strategies to prevent GvHD with immunosuppressive drugs carry significant morbidity and may affect the graft-versus-tumor (GVT) effect. Inflammatory bowel disease (IBD) is an intestinal inflammatory condition that affects more than 2 million people in the United States. Current strategies to prevent colitis with immunosuppressive drugs carry significant morbidity. Recently, Repulsive Guidance Molecule b (RGMb) has been identified as part of a signaling hub with neogenin and BMP receptors in mice and humans. In addition, RGMb binds BMP-2/4 in mice and humans as well as PD-L2 in mice. RGMb is expressed in the gut epithelium and by antigen presenting cells, and we found significantly increased expression in mouse small intestine after total body irradiation HCT conditioning. We hypothesized that RGMb may play a role in GvHD and IBD pathogenesis by contributing to mucosal inflammation. Using major-mismatched HCT mouse models, treatment with an anti-RGMb monoclonal antibody (mAb) that blocks the interaction with BMP-2/4 and neogenin prevented GvHD and improved survival compared to isotype control (75% versus 30% survival at 60 days after transplantation). The GVT effect was retained in tumor models. Using an inflammatory bowel disease dextran sulfate sodium model, treatment with anti-RGMb blocking monoclonal antibody but not isotype control prevented colitis and improved survival compared to control (73% versus 33% at 21 days after treatment) restoring gut homeostasis. Anti-RGMb mAb (9D1) treatment decreased IFN-γ and significantly increased IL-5 and IL-10 in the gut of the treated mice compared to the isotype control treated mice., Competing Interests: EM, RD, and GF are co-founders and equity holders in Triursus Therapeutics. EM is a co-founder of GigaGen, Inc. GF has patents/pending royalties on the PD-1/PD-L1 pathway from Roche, Merck MSD, Bristol-Myers-Squibb, Merck KGA, Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, and Novartis. GF has served on advisory boards for Roche, Bristol-Myers-Squibb, Xios, Origimed, Triursus, iTeos, NextPoint, IgM, Jubilant, GV20, Geode, IOME, and Invaria. GF has equity in Nextpoint, Xios, iTeos, IgM, GV20, Trillium, Invaria, and is a co-founder and equity holder in Triursus. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Pérez-Cruz, Iliopoulou, Hsu, Wu, Erkers, Swaminathan, Tang, Bader, Kambham, Xie, Dekruyff, Freeman and Meyer.)
- Published
- 2022
- Full Text
- View/download PDF
42. Immune checkpoint expression on HIV-specific CD4+ T cells and response to their blockade are dependent on lineage and function.
- Author
-
Brunet-Ratnasingham E, Morou A, Dubé M, Niessl J, Baxter AE, Tastet O, Brassard N, Ortega-Delgado G, Charlebois R, Freeman GJ, Tremblay C, Routy JP, and Kaufmann DE
- Subjects
- CD4-Positive T-Lymphocytes, Cytokines metabolism, Humans, Immune Checkpoint Inhibitors, In Situ Hybridization, Fluorescence, Programmed Cell Death 1 Receptor genetics, Programmed Cell Death 1 Receptor metabolism, RNA therapeutic use, Receptors, Chemokine metabolism, Receptors, Chemokine therapeutic use, Receptors, Immunologic metabolism, Transcription Factors metabolism, B7-H1 Antigen metabolism, HIV Infections
- Abstract
Background: Immune checkpoint blockade (ICB) partially reverses the dysfunctional state of antigen-specific T cell in chronic infections. However, its impact on the diverse subsets of CD4+ T cells in humans is largely unknown., Methods: We examined immune checkpoint (IC) expression and function in HIV-specific CD4+ T cells of viremic individuals (≥5000 vRNA cp/ml, n = 17) prior to ART and persons with spontaneous (n = 11) or therapy-induced (n = 16) viral suppression (<40 cp/ml). We investigated IC patterns associated with exhaustion-related transcription factors and chemokine receptors using activation-induced marker assays. We determined effector functions representative of T
FH , TH 1, and TH 17/TH 22 using RNA flow cytometric fluorescence in situ hybridization (FISH). We compared increase in cytokine expression upon ICB across functions and patient status., Findings: Expression of dysfunction-related molecules, such as transcription factors and ICs PD-1, TIGIT, and CD200, followed a hierarchy associated with infection status and effector profile. In vitro responsiveness to PD-L1 blockade varied with defined functions rather than IC levels: frequencies of cells with TH 1- and TH 17/TH 22-, but not TFH -related functions, increased. Cells co-expressing TH 1 and TFH functions showed response to ICB, suggesting that the cell's state rather than function dictates responsiveness to PD-L1 blockade. Response to PD-L1 blockade was strongest in viremic participants and reduced after ART initiation., Interpretation: Our data highlight a polarization-specific regulation of IC expression and differing sensitivities of antigen-specific T helper subsets to PD-1-mediated inhibition. This heterogeneity may direct and constrain ICB efficacy in restoring CD4+ T cell function in HIV infection and other diseases., Funding: NIH, CIHR, CFI, FRQS., Competing Interests: Declaration of interests G.J.F. has patents/pending royalties on the PD-1/PD-L1 pathway from Roche, Merck M.S.D., Bristol-Myers-Squibb, Merck K.G.A., Boehringer-Ingelheim, AstraZeneca, Dako, Leica, Mayo Clinic, and Novartis. G.J.F. has served on advisory boards for Roche, Bristol-Myers-Squibb, Xios, Origimed, Triursus, iTeos, and NextPoint. GJF has equity in Nextpoint, Triursus, and Xios. The anti-PD-L1 antibody BMS-936559 and the anti-TIGIT antibody BMS-g86207-Ab were given by Bristol-Myers Squibb. C.T. serves as a consultant and has received honoraria from Merck, Gilead, G.S.K., AstraZeneca and Medicago. None of aforementioned companies had implications in the design and interpretation of the experiments performed in this manuscript., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)- Published
- 2022
- Full Text
- View/download PDF
43. PD-1 combination therapy with IL-2 modifies CD8 + T cell exhaustion program.
- Author
-
Hashimoto M, Araki K, Cardenas MA, Li P, Jadhav RR, Kissick HT, Hudson WH, McGuire DJ, Obeng RC, Wieland A, Lee J, McManus DT, Ross JL, Im SJ, Lee J, Lin JX, Hu B, West EE, Scharer CD, Freeman GJ, Sharpe AH, Ramalingam SS, Pellerin A, Teichgräber V, Greenleaf WJ, Klein C, Goronzy JJ, Umaña P, Leonard WJ, Smith KA, and Ahmed R
- Subjects
- Cell Differentiation drug effects, Drug Therapy, Combination, Humans, Interleukin Receptor Common gamma Subunit, Interleukin-2 Receptor alpha Subunit, Interleukin-2 Receptor beta Subunit, Lymphocytic Choriomeningitis drug therapy, Lymphocytic Choriomeningitis immunology, T Cell Transcription Factor 1, CD8-Positive T-Lymphocytes cytology, CD8-Positive T-Lymphocytes drug effects, CD8-Positive T-Lymphocytes immunology, Interleukin-2 immunology, Interleukin-2 pharmacology, Interleukin-2 therapeutic use, Programmed Cell Death 1 Receptor antagonists & inhibitors
- Abstract
Combination therapy with PD-1 blockade and IL-2 is highly effective during chronic lymphocytic choriomeningitis virus infection
1 . Here we examine the underlying basis for this synergy. We show that PD-1 + IL-2 combination therapy, in contrast to PD-1 monotherapy, substantially changes the differentiation program of the PD-1+ TCF1+ stem-like CD8+ T cells and results in the generation of transcriptionally and epigenetically distinct effector CD8+ T cells that resemble highly functional effector CD8+ T cells seen after an acute viral infection. The generation of these qualitatively superior CD8+ T cells that mediate viral control underlies the synergy between PD-1 and IL-2. Our results show that the PD-1+ TCF1+ stem-like CD8+ T cells, also referred to as precursors of exhausted CD8+ T cells, are not fate-locked into the exhaustion program and their differentiation trajectory can be changed by IL-2 signals. These virus-specific effector CD8+ T cells emerging from the stem-like CD8+ T cells after combination therapy expressed increased levels of the high-affinity IL-2 trimeric (CD25-CD122-CD132) receptor. This was not seen after PD-1 blockade alone. Finally, we show that CD25 engagement with IL-2 has an important role in the observed synergy between IL-2 cytokine and PD-1 blockade. Either blocking CD25 with an antibody or using a mutated version of IL-2 that does not bind to CD25 but still binds to CD122 and CD132 almost completely abrogated the synergistic effects observed after PD-1 + IL-2 combination therapy. There is considerable interest in PD-1 + IL-2 combination therapy for patients with cancer2,3 , and our fundamental studies defining the underlying mechanisms of how IL-2 synergizes with PD-1 blockade should inform these human translational studies., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2022
- Full Text
- View/download PDF
44. Transcriptomic Correlates of Tumor Cell PD-L1 Expression and Response to Nivolumab Monotherapy in Metastatic Clear Cell Renal Cell Carcinoma.
- Author
-
Denize T, Hou Y, Pignon JC, Walton E, West DJ, Freeman GJ, Braun DA, Wu CJ, Gupta S, Motzer RJ, Atkins MB, McDermott D, Choueiri TK, Shukla SA, and Signoretti S
- Subjects
- B7-H1 Antigen genetics, B7-H1 Antigen metabolism, Biomarkers, Tumor metabolism, Humans, Nivolumab therapeutic use, Transcriptome, Carcinoma, Renal Cell drug therapy, Carcinoma, Renal Cell genetics, Carcinoma, Renal Cell pathology, Kidney Neoplasms drug therapy, Kidney Neoplasms genetics, Kidney Neoplasms pathology
- Abstract
Purpose: PD-L1 expression on tumor cells (TC) is associated with response to anti-PD-1-based therapies in some tumor types, but its significance in clear cell renal cell carcinoma (ccRCC) is uncertain. We leveraged tumor heterogeneity to identify molecular correlates of TC PD-L1 expression in ccRCC and assessed their role in predicting response to anti-PD-1 monotherapy., Experimental Design: RNA sequencing was performed on paired TC PD-L1 positive and negative areas isolated from eight ccRCC tumors and transcriptomic features associated with PD-L1 status were identified. A cohort of 232 patients with metastatic ccRCC from the randomized CheckMate-025 (CM-025) trial was used to confirm the findings and correlate transcriptomic profiles with clinical outcomes., Results: In both the paired samples and the CM-025 cohort, TC PD-L1 expression was associated with combined overexpression of immune- and cell proliferation-related pathways, upregulation of T-cell activation signatures, and increased tumor-infiltrating immune cells. In the CM-025 cohort, TC PD-L1 expression was not associated with clinical outcomes. A molecular RCC subtype characterized by combined overexpression of immune- and cell proliferation-related pathways (previously defined by unsupervised clustering of transcriptomic data) was enriched in TC PD-L1 positive tumors and displayed longer progression-free survival (HR, 0.32; 95% confidence interval, 0.13-0.83) and higher objective response rate (30% vs. 0%, P = 0.04) on nivolumab compared with everolimus., Conclusions: Both TC-extrinsic (immune-related) and TC-intrinsic (cell proliferation-related) mechanisms are likely intertwined in the regulation of TC PD-L1 expression in ccRCC. The quantitation of these transcriptional programs may better predict benefit from anti-PD-1-based therapy compared with TC PD-L1 expression alone in ccRCC., (©2022 American Association for Cancer Research.)
- Published
- 2022
- Full Text
- View/download PDF
45. Successful Anti-PD-1 Cancer Immunotherapy Requires T Cell-Dendritic Cell Crosstalk Involving the Cytokines IFN-γ and IL-12.
- Author
-
Garris CS, Arlauckas SP, Kohler RH, Trefny MP, Garren S, Piot C, Engblom C, Pfirschke C, Siwicki M, Gungabeesoon J, Freeman GJ, Warren SE, Ong S, Browning E, Twitty CG, Pierce RH, Le MH, Algazi AP, Daud AI, Pai SI, Zippelius A, Weissleder R, and Pittet MJ
- Published
- 2022
- Full Text
- View/download PDF
46. PD-1 blockade following ART interruption enhances control of pathogenic SIV in rhesus macaques.
- Author
-
Velu V, Titanji K, Ahmed H, Shetty RD, Chennareddi LS, Freeman GJ, Ahmed R, and Amara RR
- Subjects
- Animals, CD4-Positive T-Lymphocytes virology, CD8-Positive T-Lymphocytes virology, Immunologic Memory, Macaca mulatta, Viral Load drug effects, Viremia drug therapy, Anti-Retroviral Agents therapeutic use, Immune Checkpoint Inhibitors pharmacology, Immune Checkpoint Inhibitors therapeutic use, Programmed Cell Death 1 Receptor antagonists & inhibitors, Simian Acquired Immunodeficiency Syndrome drug therapy, Simian Acquired Immunodeficiency Syndrome immunology, Simian Immunodeficiency Virus drug effects
- Abstract
Programmed death-1 (PD-1) blockade during chronic Simian immunodeficiency virus (SIV) infection results in restoration of CD8 T-cell function and enhances viral control. Here, we tested the therapeutic benefits of PD-1 blockade administered soon after anti-retrovial therapy (ART) interruption (ATI) by treating SIV-infected and ART-suppressed macaques with either an anti-PD-1 antibody ( n = 7) or saline ( n = 4) at 4 wk after ATI. Following ATI, the plasma viremia increased rapidly in all animals, and the frequency of SIV-specific CD8 T cells also increased in some animals. PD-1 blockade post ATI resulted in higher proliferation of total memory CD8 and CD4 T cells and natural killer cells. PD-1 blockade also resulted in higher proliferation of SIV-specific CD8 T cells and promoted their differentiation toward better functional quality. Importantly, four out of the seven anti-PD-1 antibody-treated animals showed a rapid decline in plasma viremia by 100- to 2300-fold and this was observed only in animals that showed measurable SIV-specific CD8 T cells post PD-1 blockade. These results demonstrate that PD-1 blockade following ATI can significantly improve the function of anti-viral CD8 T cells and enhance viral control and strongly suggests its potential synergy with other immunotherapies that induce functional CD8 T-cell response under ART. These results have important implications for HIV cure research.
- Published
- 2022
- Full Text
- View/download PDF
47. Tumor cells dictate anti-tumor immune responses by altering pyruvate utilization and succinate signaling in CD8 + T cells.
- Author
-
Elia I, Rowe JH, Johnson S, Joshi S, Notarangelo G, Kurmi K, Weiss S, Freeman GJ, Sharpe AH, and Haigis MC
- Subjects
- CD8-Positive T-Lymphocytes metabolism, Humans, Immunity, Lactic Acid, Pyruvate Carboxylase metabolism, Succinic Acid, Tumor Microenvironment, Neoplasms, Pyruvic Acid metabolism, Pyruvic Acid pharmacology
- Abstract
The tumor microenvironment (TME) is a unique metabolic niche that can inhibit T cell metabolism and cytotoxicity. To dissect the metabolic interplay between tumors and T cells, we establish an in vitro system that recapitulates the metabolic niche of the TME and allows us to define cell-specific metabolism. We identify tumor-derived lactate as an inhibitor of CD8
+ T cell cytotoxicity, revealing an unexpected metabolic shunt in the TCA cycle. Metabolically fit cytotoxic T cells shunt succinate out of the TCA cycle to promote autocrine signaling via the succinate receptor (SUCNR1). Cytotoxic T cells are reliant on pyruvate carboxylase (PC) to replenish TCA cycle intermediates. By contrast, lactate reduces PC-mediated anaplerosis. The inhibition of pyruvate dehydrogenase (PDH) is sufficient to restore PC activity, succinate secretion, and the activation of SUCNR1. These studies identify PDH as a potential drug target to allow CD8+ T cells to retain cytotoxicity and overcome a lactate-enriched TME., Competing Interests: Declaration of interests M.C.H. and A.H.S. received research funding from Roche Pharmaceuticals. M.C.H. received funding from Agilent Technologies. M.C.H. and A.H.S. are advisers to Guided Clarity., (Copyright © 2022 Elsevier Inc. All rights reserved.)- Published
- 2022
- Full Text
- View/download PDF
48. A Comparison of Murine PD-1 and PD-L1 Monoclonal Antibodies.
- Author
-
Bu MT, Yuan L, Klee AN, and Freeman GJ
- Subjects
- Animals, Antibodies, Monoclonal, B7-H1 Antigen, Immunotherapy, Mice, Programmed Cell Death 1 Receptor, Rats, Antineoplastic Agents, Immunological, Neoplasms
- Abstract
Blockade of the PD-L1/PD-1 pathway has proven to be a broadly effective cancer immunotherapy. FDA-approved therapeutic monoclonal antibodies (mAbs) targeting the pathway have high affinity, blocking capacity, and low antibody effector activity. A number of rat antimouse mAbs have been used to model cancer immunotherapy in mouse models. We set forth the amino acid sequences of mAbs specific for mouse PD-1 (29F.1A12) and PD-L1 (10F.9G2) and compare their avidities, blocking capacities, biological activities, and epitope recognition with other commonly used mAbs. Further manipulation of these sequences should facilitate better modeling of immunotherapy in mouse models and the generation of novel agents.
- Published
- 2022
- Full Text
- View/download PDF
49. Insights into Immune Escape During Tumor Evolution and Response to Immunotherapy Using a Rat Model of Breast Cancer.
- Author
-
Gil Del Alcazar CR, Trinh A, Alečković M, Rojas Jimenez E, Harper NW, Oliphant MUJ, Xie S, Krop ED, Lulseged B, Murphy KC, Keenan TE, Van Allen EM, Tolaney SM, Freeman GJ, Dillon DA, Muthuswamy SK, and Polyak K
- Subjects
- Animals, Female, Hormones, Humans, Immunologic Factors, Immunotherapy, Mice, Rats, Rats, Sprague-Dawley, Receptors, Antigen, T-Cell, Breast Neoplasms genetics, Breast Neoplasms immunology, Breast Neoplasms therapy
- Abstract
Animal models are critical for the preclinical validation of cancer immunotherapies. Unfortunately, mouse breast cancer models do not faithfully reproduce the molecular subtypes and immune environment of the human disease. In particular, there are no good murine models of estrogen receptor-positive (ER+) breast cancer, the predominant subtype in patients. Here, we show that Nitroso-N-methylurea-induced mammary tumors in outbred Sprague-Dawley rats recapitulate the heterogeneity for mutational profiles, ER expression, and immune evasive mechanisms observed in human breast cancer. We demonstrate the utility of this model for preclinical studies by dissecting mechanisms of response to immunotherapy using combination TGFBR inhibition and PD-L1 blockade. Short-term treatment of early-stage tumors induced durable responses. Gene expression profiling and spatial mapping classified tumors as inflammatory and noninflammatory, and identified IFNγ, T-cell receptor (TCR), and B-cell receptor (BCR) signaling, CD74/MHC II, and epithelium-interacting CD8+ T cells as markers of response, whereas the complement system, M2 macrophage phenotype, and translation in mitochondria were associated with resistance. We found that the expression of CD74 correlated with leukocyte fraction and TCR diversity in human breast cancer. We identified a subset of rat ER+ tumors marked by expression of antigen-processing genes that had an active immune environment and responded to treatment. A gene signature characteristic of these tumors predicted disease-free survival in patients with ER+ Luminal A breast cancer and overall survival in patients with metastatic breast cancer receiving anti-PD-L1 therapy. We demonstrate the usefulness of this preclinical model for immunotherapy and suggest examination to expand immunotherapy to a subset of patients with ER+ disease. See related Spotlight by Roussos Torres, p. 672., (©2022 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2022
- Full Text
- View/download PDF
50. USP8 inhibition reshapes an inflamed tumor microenvironment that potentiates the immunotherapy.
- Author
-
Xiong W, Gao X, Zhang T, Jiang B, Hu MM, Bu X, Gao Y, Zhang LZ, Xiao BL, He C, Sun Y, Li H, Shi J, Xiao X, Xiang B, Xie C, Chen G, Zhang H, Wei W, Freeman GJ, Shu HB, Wang H, and Zhang J
- Subjects
- Animals, CD8-Positive T-Lymphocytes, Cell Line, Tumor, Humans, Mice, Programmed Cell Death 1 Receptor antagonists & inhibitors, Endopeptidases genetics, Endosomal Sorting Complexes Required for Transport genetics, Immunotherapy, Neoplasms drug therapy, Neoplasms genetics, Tumor Microenvironment, Ubiquitin Thiolesterase antagonists & inhibitors, Ubiquitin Thiolesterase genetics
- Abstract
Anti-PD-1/PD-L1 immunotherapy has achieved impressive therapeutic outcomes in patients with multiple cancer types. However, the underlined molecular mechanism(s) for moderate response rate (15-25%) or resistance to PD-1/PD-L1 blockade remains not completely understood. Here, we report that inhibiting the deubiquitinase, USP8, significantly enhances the efficacy of anti-PD-1/PD-L1 immunotherapy through reshaping an inflamed tumor microenvironment (TME). Mechanistically, USP8 inhibition increases PD-L1 protein abundance through elevating the TRAF6-mediated K63-linked ubiquitination of PD-L1 to antagonize K48-linked ubiquitination and degradation of PD-L1. In addition, USP8 inhibition also triggers innate immune response and MHC-I expression largely through activating the NF-κB signaling. Based on these mechanisms, USP8 inhibitor combination with PD-1/PD-L1 blockade significantly activates the infiltrated CD8
+ T cells to suppress tumor growth and improves the survival benefit in several murine tumor models. Thus, our study reveals a potential combined therapeutic strategy to utilize a USP8 inhibitor and PD-1/PD-L1 blockade for enhancing anti-tumor efficacy., (© 2022. The Author(s).)- Published
- 2022
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.