Cancer immunoprevention leverages the immune system's surveillance mechanisms to mitigate tumor development. Vaccines that constitute a tumor antigen and an immune adjuvant are perceived as immunoprevention modalities. However, relevant tumor antigens are unknown for non-viral cancers, which constitute most human cancers. Our group has recently shown that SA-4-1BBL, a novel agonist of CD137 receptor, but not antibodies, shows immunoprevention efficacy against various tumors. Advanced bioinformatics analyses of bulk RNA-seq data were conducted to elucidate mechanisms underlying cancer immunoprevention. Mice received subcutaneous injections of SA-4-1BBL or agonistic 3H3 antibody, and the injection-site tissue (IS) and draining lymph nodes (LN) were analyzed for differential gene expression. SA-4-1BBL induced a compartmentalized and temporally dynamic immune program characterized by early effector activation at IS and sustained immune regulation in draining LN. K-means clustering of 4564 DEGs identified eight functionally distinct clusters. IS-enriched clusters contained activation genes for CD4+ T and NK cells, including Cd28, Klra1, Cd4, Cd40, and Cd40l, while LN clusters were enriched for regulatory genes (Tnfaip3, Irf5, Col1a2) that ensure immune priming and homeostatic restraint for a balanced response. SA-4-1BBL generated a more selective and durable activation of adaptive immunity, TCR signaling, Th1/Th2 differentiation, and NK cytotoxicity. 3H3 activated broader innate inflammatory programs, including Toll-like receptor and neurodegeneration-linked pathways. IMPRes analysis showed that SA-4-1BBL activates sequential immune-regulatory circuits centered on Stat1, Cd247, and Ifng and modulates the CD151-TGF-β axis. These findings demonstrate that SA-4-1BBL elicits a balanced immune response, ensuring both safety and efficacy in preventing cancer development.

Allogeneic islet transplantation is an effective treatment fortype 1 diabetes, but its clinical use is limited by rejection involving innate and adaptive immune responses, requiring lifelong immunosuppression. We herein engineered islets that transiently display 2 immunomodulators chimeric with streptavidin (SA), thrombomodulin (SA-TM) and CD47 (SA-CD47), for localized modulation of both innate and adaptive immune responses. The engineering process did not impact islet viability, glucose responsiveness, and metabolic activity. Intraportal transplantation into allogeneic recipients achieved sustained survival, with 8 out of 11 grafts surviving 120-330 days without immunosuppression. In contrast, non-engineered islets were acutely rejected (median survival time [MST] = 12 days), while islets engineered with SA-TM showed delayed rejection (MST = 13.5 days) and those with SA-CD47 exhibited prolonged survival (MST = 24 days). Double-engineered islets generated a localized tolerogenic immune environment characterized by low frequencies of inflammatory innate immune cells and increased frequencies of M2 macrophages, myeloid-derived suppressor cells, and CD4+FoxP3+ T regulatory cells. The transcriptomic analysis showed downregulation of proinflammatory and upregulation of immune regulatory pathways. Our results demonstrate that transient co-display of immunomodulatory molecules on the islet surface is a versatile platform with significant translational potential for islet transplantation.

FasL and IL-2 regulate two critical signaling pathways that are essential for maintaining immune homeostasis and tolerance to self-antigens. Combined use of these molecules has significant translational potential in autoimmune diseases and transplantation. However, systemic administration of these molecules is limited by rapid clearance and potential off-target effects that can cause adverse effects. To address these limitations, we herein established an injectable, biodegradable, lipid-encapsulated condensate (LipCon) platform for sustained delivery of a novel form of FasL (SA-FasL) and IL-2 to modulate alloreactive immune responses toward regulation. This dual-delivery platform combines two complementary immunomodulatory mechanisms to promote targeted immunomodulation: where SA-FasL induces apoptosis in activated T effectors (Teffs) and IL-2 drives the expansion of regulatory T cells (Tregs). LipCon showed 99% loading efficiency for IL-2 and 65% for SA-FasL. Both proteins demonstrated sustained in vitro release over a 30-day observation period. IL-2 retained full activity throughout, whereas SA-FasL activity declined by approximately 86% by day 6. In vivo, IL-2 showed sustained release with a half-life of ∼7 days, whereas SA-FasL showed rapid loss within ∼1 day, without evidence of acute toxicity. Importantly, LipCon-mediated co-delivery of IL-2 and SA-FasL significantly reduced the frequency of T effector cells (Teffs), while increasing T regulatory cells (Tregs), thereby shifting Teff/Treg balance toward immune regulation. Collectively, these findings establish LipCon as a versatile platform having robust IL-2 retention with transient SA-FasL bioavailability showing early immunomodulatory activity and significant translational potential.

Islet transplantation can restore glycemic control in type 1 diabetes, yet the heterogeneity of patient immune responses and transplant outcomes motivates the need for technologies to monitor the graft. Since transplanted islets are not readily accessible for biopsy due to their diffuse engraftment within the liver, clinical monitoring relies on measurements such as islet mass, blood glucose, and C-peptide levels, which are lagging indicators that change only after substantial graft injury. Here, we developed a minimally invasive synthetic immunological niche (IN) that captures graft-associated immune responses through serial subcutaneous biopsy. We evaluated the IN across murine syngeneic, allogeneic, and autoimmune islet transplant models, including CD40/CD154 costimulatory blockade with anti-CD40L. In syngeneic versus allogeneic recipients, IN identified immune populations and transcriptomic signatures that mirrored the graft and distinguished healthy from rejecting grafts. In anti-CD40L treated allografts, IN revealed innate macrophage- and dendritic cell-associated programs linked to graft acceptance versus rejection, whereas IN from untreated allografts showed stronger adaptive immune signatures. Longitudinal IN profiling further detected progressive inflammatory activation in accepted allografts, indicating persistent subclinical risk. Finally, in an autoimmune allograft model treated with anti-CD40L plus rapamycin, IN identified a 13-gene signature that separated early from late rejection trajectories and distinguished autoimmune- from alloimmune-associated rejection programs. Overall, these findings establish IN as a surrogate tissue for minimally invasive monitoring of islet graft and early detection of rejection-associated immune dysregulation.

A novel CD137 agonist, SA-4-1BBL, as a single agent is effective in training the immune system to prevent cancer development in preclinical models. The cancer immunoprevention efficacy of SA-4-1BBL is tumor-type independent, long-lasting (>14 weeks), and requires immune cells expressing CD4 or NK1.1 molecules but not CD8+ T cells, implicating innate immune surveillance. Since NKT cells express both CD4 and NK1.1, we herein investigated the role of these cells, phagocytes, and neutrophils in cancer immunoprevention efficacy of SA-4-1BBL. Wild-type, CD1d-/-KO lacking all NKT cells, and Jα18-/-KO lacking invariant NKT cells were treated subcutaneously with SA-4-1BBL twice two weeks apart. Mice were challenged two weeks later subcutaneously with syngeneic cervical cancer (TC-1) or Lewis lung carcinoma (LLC) and monitored for tumor development. To deplete phagocytes, clodronate-loaded liposomes were administrated 2 days prior to each SA-4-1BBL treatment (priming) or 2 days before the tumor challenge (effector). Neutrophils were depleted using anti-Ly6G Ab 2 days before each SA-4-1BBL treatment. Mice treated with PBS-loaded liposome or saline served as controls. Draining lymph nodes from tumor-free mice at 60 days post-challenge were used for deep immunophenotyping to assess long-lasting immune correlates of cancer immunoprevention. Pretreatment with SA-4-1BBL prevented tumor growth in both TC-1 and LLC challenged mice (> 70%). Deep immunophenotyping demonstrated that SA-4-1BBL significantly increased absolute numbers of CD4+ T, NK, NKT cells, macrophages, and neutrophils as compared to naïve mice. SA-4-1BBL was effective in preventing tumor growth in mice lacking invariant NKT (Jα18-/-KO, 100%), all NKT (CD1d-/-KO, >60%) cells, or mice depleted for neutrophils (>70%). Importantly, clodronate-mediated phagocyte depletion significantly reduced the tumor immunoprevention efficacy to 20% at both priming and effector phases, whereas treatment with PBS-loaded liposome as control showed a similar immunoprevention efficacy (∼68%) as SA-4-1BBL-treated experimental group. These studies demonstrated that phagocytes (monocytes, macrophages, and dendritic cells) but not NKT or neutrophils play critical roles in cancer immunoprevention efficacy of SA-4-1BBL. Ongoing studies using mice lacking monocyte/macrophage or DCs will elucidate the contribution of each cell population to the immunoprevention efficacy of SA-4-1BBL. Elucidation of mechanistic underlying of cancer immunoprevention will facilitate clinical translation of this approach as a potential universal platform to combat cancer. Acknowledgment: Supported in part by Paula and Rodger Riney Foundation and NCI (1UG3CA290305). Feyza Nur Arguc, Mohammad Tarique, Vahap Ulker, Esma S. Yolcu, Haval Shirwan. Phagocytes play a critical role in the efficacy of a novel agonist of CD137 in preventing various tumor types [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_2):Abstract nr LB390.

Cigarette smoke exposure (CSE) increases the risk for a plethora of cancers. Recent evidence indicates that the gut microbiome can influence cancer progression by immune system modulation. Since CSE alters the gut microbiome, we hypothesized that the gut microbiome serves as a causative link between smoking and cancer growth. Through a combination of syngeneic animal models and fecal microbiota transplantation studies, we established an essential role for smoke-induced dysbiosis in cancer growth. 16s rRNA sequencing and liquid chromatography-mass spectrometry indicated a unique CSE-associated microbial and metabolomic signature. Immunophenotyping of tumor specimens and experiments in Rag1-KO and CD8-KO demonstrated that smoke-induced tumor growth requires functional adaptive immunity. Finally, utilizing gut microbial ablation strategies with broad- and narrow-spectrum antibiotics, we demonstrated the reversal of phenotypic effects of CSE. Our study provides evidence for gut microbiome as an actionable target to mitigate CSE-induced tumor promotion.

Liver cancer, particularly Hepatocellular Carcinoma (HCC), remains a significant global health challenge owing to its high incidence and position as the fourth leading cause of cancer-related mortality. HCC represents 75-85% of all liver cancer cases and ranks as the sixth most prevalent cancer globally. Several factors, including late-stage diagnosis, limited treatment effectiveness, resistance to conventional therapies, and adverse side effects, hinder the delivery of life-prolonging care to patients with HCC. Current treatment options such as chemotherapy, immunotherapy, and adjuvant therapy are often associated with severe side effects. Consequently, there is an urgent need for improved diagnostic methods and alternative therapeutic approaches to extend life expectancy and reduce HCC-related mortalities. Artificial Intelligence (AI) is an emerging technology that offers promising advances for the early detection of HCC. In terms of alternative treatments, natural compounds have garnered significant attention because of their diverse biological activities, such as antitumor, antiviral, antimicrobial, antioxidant, anti-inflammatory, hepatoprotective, antimutagenic, and cardioprotective effects, and their relatively lower side effect profiles. These compounds exhibit hepatoprotective properties by modulating key molecular pathways involved in HCC development and progression. This article provides an overview of recent advances in the understanding of liver cancer etiology, therapeutic targets in HCC pathogenesis, the role of AI in its detection, and the potential of natural products, particularly flavonoids, as preventive and therapeutic agents against HCC, highlighting their underlying mechanisms of action.

Pancreatic cancer is one of the most lethal malignancies of the digestive tract, with a poor prognosis and a 5-year survival rate of less than 10%. The highly aggressive nature of pancreatic cancer results in a mortality rate of approximately 50% within the 6-month period of diagnosis. The absence of disease-specific symptoms significantly impedes early detection and timely intervention, contributing to its high mortality rate. Current treatment options such as radiotherapy, chemotherapy, and surgery are often inadequate for complete disease eradication and are associated with severe side effects that compromise patients' overall health. As a result, there is an urgent need for novel therapeutic strategies to address the rapidly increasing incidence of pancreatic cancer while ensuring safer, cost and more effective treatment alternatives. Plant-derived polyphenols have emerged as promising candidates due to their potent anticancer properties and minimal side effects compared to conventional therapies. In this review, we explore the biological significance and anticancer mechanisms of key polyphenols, including quercetin, resveratrol, apigenin, luteolin, EGCG, and curcumin, with a particular focus on their role in combating pancreatic cancer. Additionally, we provide a comprehensive summary of various pancreatic cancer studies, including ongoing clinical trials from the past decade.

Abstract Introduction: We recently demonstrated that pretreatment of mice with SA-4-1BBL, a novel oligomeric form of CD137 agonist, as a single agent prevents the development of various tumor types in mice. The tumor immunoprevention efficacy is long-lasting (>14 weeks) and contingent upon CD4+ immune cells, NK cells, and IFN-γ, but not CD8+ T and B cells. Trained immunity mediated by previously activated myeloid cells has been shown to have efficacy against cancer. The objective of this study was twofold: i) assess the role of trained immunity and ii) elucidate the mechanistic underlying of cancer immunoprevention. Study Design: Wild-type C57BL/6 mice and various transgenic (CD4+ OT-II.Rag+/+, CD4+ OT-II.Rag−/−, CD8+ OT-I/Rag2−/- KO, and Rag1−/−) mice on C57BL/6 background were pretreated subcutaneously with SA-4-1BBL or an agonistic Ab that does not have immunoprevention efficacy (3H3) twice two weeks apart. Mice were challenged two weeks later with a cervical cancer cell line (TC-1) and monitored for tumor development. A group of mice was also treated with monophosphoryl lipid A (MPLA), an agonist of TLR4 involved in trained immunity, using the above scheme. Mice treated with saline served as the control throughout all studies. Deep immunophenotyping was conducted to assess immune correlates of cancer immunoprevention. Results: Pretreatment with SA-4-1BBL prevented tumor growth in wild-type mice (> 60%), whereas all MPLA treated mice developed tumor. SA-4-1BBL did not show tumor immunoprevention efficacy in mice lacking adaptive immune cells (Rag1−/- KO), OT-II cells on Rag−/- or Rag+/+ background, and OT-I on Rag2−/− background. Deep immunophenotyping revealed a significant increase in the absolute number of CD4+ T central memory and CD4+ T stem cell-like memory cells, NK cells, NKT cells, and various myeloid cells in mice treated with SA-4-1BBL as compared with those treated with the 3H3 Ab or saline treated controls. Conclusions: These results demonstrate that cancer immunoprevention efficacy of SA-4-1BBL requires a diverse T cell receptor repertoire as OT-II mice on Rag+/+ background, restricted to a TCRβ chain, but have all other myeloid and NK cells, do not show efficacy. Furthermore, treatment with MPLA involved in trained immunity has no efficacy. Altogether, these observations do not implicate trained immunity as a mechanism of cancer immunoprevention. Elucidation of the mechanistic basis of SA-4-1BBL immunoprevention may have significant clinical implications. Citation Format: Feyza Nur Arguc, Mohammad Tarique, Vahap Ulker, Esma S. Yolcu, Haval Shirwan. A novel agonist of CD137 as a single agent shows cancer immunoprevention efficacy against various tumor types that is not dependent on the trained immunity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2465.

CAR T cell therapy has revolutionized the cancer therapy. However, for an effective therapy, selecting an ideal target antigen on the cancer cell is a prerequisite. The purpose of this review is to provide an overview of all the possible cancer antigens in hematological malignancies that can be targeted by CAR T cells. Owing to the fast-paced research in the field of CAR T cell therapy, a plethora of surface antigens on the cancer cell have been identified against which CAR T cells have been successfully made. Over the years, CAR T cells targeting CD19 and BCMA have been approved by the FDA, and many others have shown tremendous efficacy to combat cancer. This review gives an overview of all the cancer antigens that have been targeted by CAR T cell therapy.

Natural 4-1BBL (CD137L) is a cell membrane-bound protein critical to the expansion, effector function, and survival of CD8+ T cells. We reported the generation of an active soluble oligomeric construct, SA-4-1BBL, with demonstrated immunoprevention and immunotherapeutic efficacy in various mouse tumor models. Herein, we developed an oncolytic adenovirus (OAd) for the delivery and expression of SA-4-1BBL (OAdSA-4-1BBL) into solid tumors for immunotherapy. SA-4-1BBL protein expressed by this construct produced T-cell proliferation in vitro. OAdSA-4-1BBL decreased cell viability in two mouse lung cancer cell lines, TC-1 and CMT64, but not in the non-cancerous lung MM14.Lu cell line. OAdSA-4-1BBL induced programmed cell death types I and II (apoptosis and autophagy, respectively), and autophagy-mediated adenosine triphosphate (ATP) release was also detected. Intratumoral injection of OAdSA-4-1BBL efficiently expressed the SA-4-1BBL protein in the tumors, resulting in significant tumor suppression in a syngeneic subcutaneous TC-1 mouse lung cancer model. Tumor suppression was associated with a higher frequency of dendritic cells and an increased infiltration of cytotoxic CD8+ T and NK cells into the tumors. Our data suggest that OAdSA-4-1BBL may present an efficacious alternative therapeutic strategy against lung cancer as a standalone construct or in combination with other immunotherapeutic modalities, such as immune checkpoint inhibitors.

Abstract Allogeneic islet transplantation is an effective treatment for type 1 diabetes (T1D). Graft rejection is controlled by immunosuppressive drugs that have adverse effects. An imbalance between Teffs and Tregs is the primary cause of T1D and graft rejection. Activated Teff cells express Fas and are sensitive to FasL-mediated apoptosis. Treg cells are relatively refractive to apoptosis and expand in response to IL-2. Thus, a combination of Fas and IL-2R agonists has the potential to modulate alloreactive responses for sustained graft survival. Given the off-target effects of systemic delivery of biologics, we established a lipocoacervate formulation consisting of a biodegradable polycation, PEAD, and polyanion heparin to form coacervate for controlled delivery of a novel Fas agonist, SA-FasL, and IL-2. Release kinetics and activities of proteins were assessed in vitro. The immunomodulatory efficacy of protein-loaded lipocoacervate was assessed in vivo. IL-2 and SA-FasL proteins showed a steady release over 30 days in vitro. IL-2 released on day 14 had minimal activity loss, whereas FasL showed ~20% activity loss on day 9. Treatment with SA-FasL/IL-2-loaded lipocoacervate modulated in vivo alloreactive T cell responses, resulting in an increased Treg/Teff cell ratio. In conclusion, lipocoacervate is an effective platform for controlled and sustained release of SA-FasL and IL-2 biologics to modulate allo and autoreactive T cells with significant therapeutic potential for T1D.

Chronic pancreatitis (CP) is a fibro-inflammatory disease of the pancreas with no specific cure. Research highlighting the pathogenesis and especially the therapeutic aspect remains limited. Aberrant activation of developmental pathways in adults has been implicated in several diseases. Hedgehog pathway is a notable embryonic signaling pathway, known to promote fibrosis of various organs when overactivated. The aim of this study is to explore the role of the hedgehog pathway in the progression of CP and evaluate its inhibition as a novel therapeutic strategy against CP. CP was induced in mice by repeated injections of l-arginine or caerulein in two separate models. Mice were administered with the FDA-approved pharmacological hedgehog pathway inhibitor, vismodegib during or after establishing the disease condition to inhibit hedgehog signaling. Various parameters of CP were analyzed to determine the effect of hedgehog pathway inhibition on the severity and progression of the disease. Our study shows that hedgehog signaling was overactivated during CP and its inhibition was effective in improving the histopathological parameters associated with CP. Vismodegib administration not only halted the progression of CP but was also able to resolve already-established fibrosis. In addition, inhibition of hedgehog signaling resulted in the reversal of pancreatic stellate cell activation ex vivo. Findings from our study justify conducting clinical trials using vismodegib against CP and, thus, could lead to the development of a novel therapeutic strategy for the treatment of CP. NEW & NOTEWORTHY Hedgehog signaling is activated in human and experimental models of CP. Inhibition of hedgehog signaling using an FDA-approved inhibitor, vismodegib, leads to the resolution of fibrosis and improves CP. This study has immense and immediate translational benefits.

Most pancreatic islets are destroyed immediately after intraportal transplantation by an instant blood-mediated inflammatory reaction (IBMIR) generated through activation of coagulation, complement, and proinflammatory pathways. Thus, effective mitigation of IBMIR may be contingent on the combined use of agents targeting these pathways for modulation. CD47 and thrombomodulin (TM) are two molecules with distinct functions in regulating coagulation and proinflammatory responses. We previously reported that the islet surface can be modified with biotin for transient display of novel forms of these two molecules chimeric with streptavidin (SA), that is, thrombomodulin chimeric with SA (SA-TM) and CD47 chimeric with SA (SA-CD47), as single agents with improved engraftment following intraportal transplantation. This study aimed to test whether islets can be coengineered with SA-TM and SA-CD47 molecules as a combinatorial approach to improve engraftment by inhibiting IBMIR. Mouse islets were effectively coengineered with both molecules without a detectable negative impact on their viability and metabolic function. Coengineered islets were refractory to destruction by IBMIR ex vivo and showed enhanced engraftment and sustained function in a marginal mass syngeneic intraportal transplantation model. Improved engraftment correlated with a reduction in intragraft innate immune infiltrates, particularly neutrophils and M1 macrophages. Moreover, transcripts for various intragraft procoagulatory and proinflammatory agents, including tissue factor, HMGB1 (high-mobility group box-1), IL-1β, IL-6, TNF-α, IFN-γ, and MIP-1α, were significantly reduced in coengineered islets. These data demonstrate that the transient codisplay of SA-TM and SA-CD47 proteins on the islet surface is a facile and effective platform to modulate procoagulatory and inflammatory responses with implications for both autologous and allogeneic islet transplantation.