CTLA-4 Inhibitors Research Articles

A plasma-based proteomic platform for predicting clinical benefit from immune checkpoint inhibitors in multiple cancers.

2568 Background: Immune checkpoint inhibitor (ICI)-based therapies are preferred treatment options for multiple cancer types. However, there is an unmet need for predictive tests that can identify patients likely to benefit. PROphet-NSCLC is a commercially available pretreatment plasma proteomic test that predicts clinical benefit from first-line PD-(L)1 inhibitor-based therapy in patients with metastatic non-small cell lung cancer (NSCLC), guiding the choice between ICI monotherapy versus ICI-chemotherapy. Here, we evaluate the broader clinical utility of PROphet-NSCLC by testing its predictive capabilities across diverse cancer types. Methods: Pre-treatment plasma samples and clinical data were collected for an observational study from patients with metastatic melanoma (n=68) and HPV-related cancers (n=43; NCT03427411) treated with PD-(L)1 inhibitor-based therapies. HPV-related cancers included anogenital squamous cell carcinoma (n=16), cervical carcinoma (n=18), and head and neck squamous cell carcinoma (n=9). Patients were assigned a PROphet-positive or -negative result based on computational analysis of SomaScan-derived proteomic profiles in pre-treatment plasma samples. Overall survival (OS) and progression-free survival (PFS) in PROphet-positive- and -negative groups was analyzed with the Kaplan-Meier method. Hazard ratios (HR) were calculated from univariate and multivariate Cox proportional hazard models. Results: In the melanoma cohort, PROphet-positive patients (n=51) displayed a significant OS benefit in comparison to PROphet-negative patients (n=17; median OS 92.8 months vs. 9.5 months, HR=0.14, 95% CI: 0.06-0.34, p<0.0001), consistent with the clinically validated predictive performance of the test in patients with NSCLC. OS results remained significant after correcting for sex, age, histology, ECOG performance status, and treatment type (HR=0.05, 95% CI: 0.01-0.25, p<0.001). In PROphet-positive patients, PD-1+CTLA-4 inhibitor combination therapy (n=27) was superior to PD-1 inhibitor monotherapy (n=24; OS: median 118.4 vs. 48.3 months, HR=0.58, p=0.24; PFS: median not reached vs. 10.8 months, HR=0.48, p=0.04). PROphet-negative patients displayed similarly poor outcomes with either treatment, providing a rationale to consider alternative therapies for such patients. In the HPV-related cohort, the median OS in PROphet-positive (n=10) vs PROphet-negative (n=33) groups was 43.6 vs 4.4 months (HR=0.22, 95% CI: 0.08-0.59, p=0.001), with similar trends per sub-cohort. Conclusions: Our findings show that the PROphet-NSCLC test can be applied to PD-(L)1 inhibitor-treated melanoma and HPV-related cancers, suggesting applicability to cancers beyond NSCLC. Analysis of additional patient samples is needed to explore the potential utility of PROphet-NSCLC for informing treatment decisions for a broad range of cancer types.

Comparing rates of immune checkpoint inhibitor toxicity in different treatment settings (neoadjuvant, adjuvant, and palliative) in non-small cell lung cancer (NSCLC): A meta-analysis of phase III clinical trials.

8645 Background: Immunotherapy has created a paradigm shift in the treatment of NSCLC. There is growing interest in the utility of immunotherapy alone or in combination with chemotherapy for NSCLC in the peri-operative setting with practice changing results demonstrated in several pivotal phase III clinical trials. Nevertheless, immune related adverse events (irAEs) are the main barrier in the care of patients treated with immune checkpoint inhibitors (ICIs) with grade ≥ 3 irAEs seen in 10-15% of patients. The present meta-analysis aims to compare rates of irAEs across different treatment settings in NSCLC to aid in individual patient treatment decisions. Methods: We extracted data from selected published phase III clinical trials which have led to the approval of PD-1/PDL-1 ICIs in the neoadjuvant, adjuvant, peri-operative and in the palliative setting. We excluded trials which did not have supplemental data on irAEs included with published results and trials of CTLA-4 inhibitors. A sensitivity analysis was performed to assess whether number of cycles correlated with toxicity rates. Results: Pooled data from 15 published phase III NSCLC clinical trials including 5918 patients were included. This included 1 neoadjuvant, 2 peri-operative, 3 adjuvant, and 9 palliative studies. One of the peri-operative trials included data on irAEs which occurred during the neoadjuvant/surgery phase only and this data was pooled within the neoadjuvant group. The ICI drugs included nivolumab, pembrolizumab, atezolizumab, durvalumab, cemiplimab +/- chemotherapy. This included a total of 2523 stage 1B-3C and 3395 stage IV patients. We found a 9.5% rate of grade 3-5 and 35.8% rate of all grade irAEs in palliative trials (n=3395). In contrast, neoadjuvant ICI (n=572) had a 3.5% rate of grade 3-5 and 16.8% of all grades irAEs. Peri-operative ICI (n=797) had a 5.4% rate of grade 3-5 and 24.47% all grades irAEs. Adjuvant ICI (n=1550) had a 7% rate of grade 3-5 and 38.52% rate of all grade irAEs. The relative risk (RR) between the palliative and the neoadjuvant setting of a grade 3-5 irAE is 2.35 (95% CI:1.54 to 3.59; p = 0.0001) and the RR of any irAE is 2.03 (95% CI: 1.69 to 2.43; p=0.0001). The following median number of ICI cycles was seen: neoadjuvant = 3.5, adjuvant = 18, peri-operative = 18, palliative = 10. Lower number of cycles did correlate with lower toxicity in the neoadjuvant setting but this was not seen in other treatment settings. Conclusions: Neoadjuvant, peri-operative and adjuvant ICI’s are associated with a lower rate of irAEs than that seen in palliative trials. A neoadjuvant-only approach is associated with the lowest rates of irAEs and this may be due to the lower number of cycles received. These findings have implications for individual patient treatment decisions and support the use of neoadjuvant immunotherapy in NSCLC.

Recent Advances in Nanomaterial-Based Immunotherapy for Prostate Cancer: A Comprehensive Review

Abstract: Prostate Cancer (PCa) remains a global health concern, and recent advancements in nanomaterial- based immunotherapy are reshaping the landscape of its treatment. The advent of Prostate- Specific Antigen (PSA) screening had a significant impact on the PCa burden until the early 21st century, but the ongoing innovations in therapeutic approaches and early detection methods have contributed to a decline in mortality rates. This comprehensive review delves into the evolving role of the immune system in cancer, with a particular emphasis on the latest advances in nanomaterial- based immunotherapy for PCa. The review focuses on the burgeoning field of nanomaterial- based immunotherapy for PCa, particularly in the context of vaccine-based therapies and immune checkpoint inhibitors. Recent developments in clinical trials highlight the effectiveness of immune checkpoint inhibitors, such as CTLA-4 and PD-1 inhibitors, either as standalone treatments or in combination for metastatic Castration-Resistant Prostate Cancer (mCRPC). We highlight ongoing clinical trials that explore PSMA-targeted CAR-T cells for mCRPC patients, offering valuable insights into the promising field of nanomaterial-based immunotherapy. The review also discusses nanomaterial-based vaccine treatments, such as Sipuleucel-T (Provenge®) and G-VAX. These work in different ways to boost the immune system's response to Tumor-Associated Antigens (TAAs). We also explore viral vector-based vaccines and gene therapy approaches, highlighting their potential to enhance the immune system's ability to target prostate cancer cells at the nanoscale. The article concludes with an in-depth discussion of the current and emerging nanomaterial-based biomarkers for PCa diagnosis and prognosis.

Sarcomatoid and Rhabdoid Renal Cell Carcinoma: Clinical, Pathologic, and Molecular Genetic Features.

Renal cell carcinoma (RCC) with sarcomatoid and rhabdoid morphologies has an aggressive biological behavior and a typically poor prognosis. The current 2022 WHO classification of renal tumors does not include them as distinct histologic entities but rather as transformational changes that may arise in a background of various distinct histologic types of RCC. The sarcomatoid component shows malignant spindle cells that may grow as intersecting fascicles, which is reminiscent of pleomorphic undifferentiated sarcoma. The rhabdoid cells are epithelioid cells with eccentrically located vesicular nuclei with prominent nucleoli and large intracytoplasmic eosinophilic inclusions. Studies have shown that RCCs with sarcomatoid and rhabdoid differentiation have distinctive molecular features. Sarcomatoid RCC harbors shared genomic alterations in carcinomatous and rhabdoid components, but also enrichment of specific genomic alterations in the sarcomatoid element, suggesting molecular pathways for development of sarcomatoid growth from a common clonal ancestor. Rhabdoid differentiation also arises through clonal evolution although less is known of specific genomic alterations in rhabdoid cells. Historically, treatment has lacked efficacy, although recently immunotherapy with PD-1/PD-L1/CTLA-4 inhibitors has produced significant clinical responses. Reporting of sarcomatoid and rhabdoid features in renal cell carcinoma is required by the College of American Pathologists and the International Collaboration on Cancer Reporting. This manuscript reviews the clinical, pathologic, and molecular features of sarcomatoid RCC and rhabdoid RCC with emphasis on the morphologic features of these tumors, significance of diagnostic recognition, the molecular mechanisms of tumorigenesis and differentiation along sarcomatoid and rhabdoid lines, and advances in treatment, particularly immunotherapy.

Transient thyrotoxicosis and hypothyroidism after combination therapy with pd-1 and ctla-4 inhibitors for cervical cancer

Transient thyrotoxicosis and hypothyroidism after combination therapy with pd-1 and ctla-4 inhibitors for cervical cancer

Chordoma Genetic Aberrations and Targeted Therapies Panorama: A Systematic Literature Review.

Background: Chordomas pose a challenge in treatment due to their local invasiveness, high recurrence, and potential lethality. Despite being slow-growing and rarely metastasizing, these tumors often resist conventional chemotherapies (CTs) and radiotherapies (RTs), making surgical resection a crucial intervention. However, achieving radical resection for chordomas is seldom possible, presenting therapeutic challenges. The accurate diagnosis of these tumors is vital for their distinct prognoses, yet differentiation is hindered by overlapping radiological and histopathological features. Fortunately, recent molecular and genetic studies, including extracranial location analysis, offer valuable insights for precise diagnosis. This literature review delves into the genetic aberrations and molecular biology of chordomas, aiming to provide an overview of more successful therapeutic strategies. Methods: A systematic search was conducted across major medical databases (PubMed, Embase, and Cochrane Library) up to 28 January 2023. The search strategy utilized relevant Medical Subject Heading (MeSH) terms and keywords related to "chordomas", "molecular biology", "gene aberrations", and "target therapies". The studies included in this review consist of preclinical cell studies, case reports, case series, randomized controlled trials, non-randomized controlled trials, and cohort studies reporting on genetic and biological aberrations in chordomas. Results: Of the initial 297 articles identified, 40 articles were included in the article. Two tables highlighted clinical studies and ongoing clinical trials, encompassing 18 and 22 studies, respectively. The clinical studies involved 185 patients diagnosed with chordomas. The tumor sites were predominantly sacral (n = 8, 44.4%), followed by clivus (n = 7, 38.9%) and lumbar spine (n = 3, 16.7%). Primary treatments preceding targeted therapies included surgery (n = 10, 55.6%), RT (n = 9, 50.0%), and systemic treatments (n = 7, 38.9%). Various agents targeting specific molecular pathways were analyzed in the studies, such as imatinib (a tyrosine kinase inhibitor), erlotinib, and bevacizumab, which target EGFR/VEGFR. Common adverse events included fatigue (47.1%), skin reactions (32.4%), hypertension (23.5%), diarrhea (17.6%), and thyroid abnormalities (5.9%). Clinical outcomes were systematically assessed based on progression-free survival (PFS), overall survival (OS), and tumor response evaluated using RECIST or CHOI criteria. Notably, stable disease (SD) occurred in 58.1% of cases, and partial responses (PRs) were observed in 28.2% of patients, while 13.7% experienced disease progression (PD) despite targeted therapy. Among the 22 clinical trials included in the analysis, Phase II trials were the most prevalent (40.9%), followed by I-II trials (31.8%) and Phase I trials (27.3%). PD-1 inhibitors were the most frequently utilized, appearing in 50% of the trials, followed by PD-L1 inhibitors (36.4%), CTLA-4 inhibitors (22.7%), and mTOR inhibitors (13.6%). Conclusions: This systematic review provides an extensive overview of the state of targeted therapy for chordomas, highlighting their potential to stabilize the illness and enhance clinical outcomes.

Abstract 107: Effects Of Immune-checkpoint Inhibitors On Atherosclerosis In Halted Progression Model

Immune checkpoint inhibitor (ICI) treatment in cancer has been linked to atherosclerotic-related cardiovascular events. While ICIs have been shown to aggravate atherosclerosis in mouse models of plaque progression, these models do not reflect the clinical scenario of cancer patients with pre-existing atherosclerosis under lipid-lowering treatments. To address this, we tested the effects of PD1 or CTLA4 inhibitors (aPD1 or aCTLA4), and their combination (aPD1 + aCTLA4), in a model of halted atherosclerosis progression, mimicking standard of care lipid-lowering treatments. Ldlr -/- mice (n=126) mice were fed a western diet (WD) for 17 wk (baseline), followed by a switch to standard chow diet for 4 wk (halted progression), and mice were randomized to treatment arms. Histological and single cell RNA sequencing (scRNA-seq) revealed that inhibiting PD-1 or CTLA-4 individually aggravated atherosclerosis, despite comparable cholesterol reduction to isotype control groups. Surprisingly, the combination of aPD1 + aCTLA4 did not show the same proatherogenic effect observed individually. Distinct effects on plaque composition were noted, with aPD-1 or aCTLA4 leading to increased plaque collagen content, and their combination showing plaque stabilization with higher cap thickness and similar collagen content to controls. Immunostaining revealed no differences in plaque macrophage content among groups, but aPD-1-treated mice exhibited increased CD8 T cells. scRNAseq showed that aPD1 increased specific inflammatory myeloid clusters, including Spp1+ macrophages and Ccr2+ MoDCs, and reduced anti-inflammatory IL16+ DPTs but increased cytotoxic Gzmk+ CD8+ EMRA T cells. CTLA4 inhibition increased foamy Trem2hi macrophages and Ccr2+ MoDCs. Yet, combined aPD1 + aCTLA4 did not significantly alter cell proportions compared to halted progression. In conclusion, our study demonstrates that inhibiting either PD1 or CTLA4 individually aggravates atherosclerosis, under cholesterol lowering conditions. These findings highlight the nuanced effects of immune checkpoint inhibitors on atherosclerosis, emphasizing the potential importance of considering combination therapies and lipid lowering for better outcomes in cancer survivors.

HIV-derived Proteins induce Hypertension, Sympatho-activation, and neuroinflammation via T cell-dependent mechanisms

Combination antiretroviral therapy (cART) has markedly increased life expectancy in people living with HIV (PLWH) and induced a switch in the cause of death from opportunistic diseases to cardiovascular disease (CVD). CVD is currently the leading cause of death in PLWH and hypertension, the primary risk factor for CVD, presents in more than 65% of them. However, the mechanisms whereby hypertension develops in PLWH remain unknown. In PLWH on cART, T lymphocytes (T cells) remain a viral reservoir for HIV and continue to secrete viral proteins. Viral proteins remain in circulation and are expressed in the central nervous system. Herein, we tested the hypothesis that expression of HIV-derived proteins increases blood pressure (BP), induces sympatho-activation and inflammation of the central nervous system. We utilized the Tg26 mouse model which expresses 7/9 viral proteins. Quantitative PCR analysis of viral protein expression showed that Tat, gp120, Nef, Vif, and Vpr are primarily expressed in spleen, lung, and brain tissues in Tg26 mice. Morphometric analysis revealed slight but significant increases and decreases in lean mass and fat weights respectively in Tg26. Flow cytometry analysis showed no reduction in CD4+ T cells in Tg26 mice indicating that this mouse model does not progress to AIDS and reproduce the conditions of virally suppressed PLWH. Blood pressure (BP) measurement via telemetry revealed a hypertensive phenotype in male and female Tg26 mice. Hypertension was associated with a larger drop in BP in response to ganglionic blockade but no alterations in heart rate responses to muscarinic and β-adrenergic receptor blockade in Tg26 mice, indicating that viral proteins expression increases neurogenic control of BP but did not alter cardiac autonomic control. β-adrenergic receptor blockade did not reduce BP in Tg26 mice. Tg26 mice also display increased aortic vasoconstriction to phenylephrine. To test the contribution of T cells in BP regulation and sympatho-activation, mice were subjected to a CTLA-4 inhibitor, abatacept, which inhibits T cell activation. Abatacept treatment significantly reduced BP, sympatho-activation, and phenylephrine constriction in Tg26 mice with no effects on wild-type mice. To determine whether the increase in sympatho-activation involves central inflammation, inflammatory cytokines and T cell markers levels were measured in the brain. Levels of pro-inflammatory cytokines including IL-1α, TNFα, IL-6, and CCL2 were increased in the brain of Tg26 mice as well as levels of the T-cell marker GATA3. Altogether, our data shows that the expression of HIV-derived proteins increases BP, sympatho-activation, and vascular adrenergic contractility via T cell-dependent mechanisms likely through central mechanisms involving neuro-inflammation. R01HL147639-01A1, R01HL15526501 and 19EIA34760167 (E.J.B). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Real-world evidence of efficacy of pembrolizumab plus chemotherapy and nivolumab plus ipilimumab plus chemotherapy as initial treatment for advanced non-small cell lung cancer.

For advanced non-small cell lung cancer (NSCLC), combination therapies including a PD-1 inhibitor plus chemotherapy or a PD-1 inhibitor, CTLA-4 inhibitor, and chemotherapy are standard first-line options. However, data directly comparing these regimens are lacking. This study compared the efficacy of pembrolizumab plus chemotherapy (CP) against nivolumab plus ipilimumab and chemotherapy (CNI) in a real-world setting. In this multicenter retrospective study, we compared the efficacy and safety of CP and CNI as first-line therapies in 182 patients with stage IIIB-IV NSCLC. Primary outcomes were overall survival (OS) and progression-free survival (PFS), while secondary outcomes included the response rate (RR) and safety profiles. Kaplan-Meier survival curves and Cox proportional hazards models were utilized for data analysis, adjusting for confounding factors such as age, gender, and PD-L1 expression. In this study, 160 patients received CP, while 22 received CNI. The CP group was associated with significantly better PFS than the CNI group (median 11.7 vs. 6.6 months, HR 0.56, p = 0.03). This PFS advantage persisted after propensity score matching to adjust for imbalances. No significant OS differences were observed. Grade 3-4 adverse events occurred comparably, but immune-related adverse events were numerically more frequent in the CNI group. In real-world practice, CP demonstrated superior PFS compared with CNI. These findings can inform treatment selection in advanced NSCLC.

Immune-Related Adverse Events Due to Cancer Immunotherapy: Immune Mechanisms and Clinical Manifestations.

The landscape of cancer treatment has undergone a significant transformation with the introduction of Immune Checkpoint Inhibitors (ICIs). Patients undergoing these treatments often report prolonged clinical and radiological responses, albeit with a potential risk of developing immune-related adverse events (irAEs). Here, we reviewed and discussed the mechanisms of action of ICIs and their pivotal role in regulating the immune system to enhance the anti-tumor immune response. We scrutinized the intricate pathogenic mechanisms responsible for irAEs, arising from the evasion of self-tolerance checkpoints due to drug-induced immune modulation. We also summarized the main clinical manifestations due to irAEs categorized by organ types, detailing their incidence and associated risk factors. The occurrence of irAEs is more frequent when ICIs are combined; with neurological, cardiovascular, hematological, and rheumatic irAEs more commonly linked to PD1/PD-L1 inhibitors and cutaneous and gastrointestinal irAEs more prevalent with CTLA4 inhibitors. Due to the often-nonspecific signs and symptoms, the diagnosis of irAEs (especially for those rare ones) can be challenging. The differential with primary autoimmune disorders becomes sometimes intricate, given the clinical and pathophysiological similarities. In conclusion, considering the escalating use of ICIs, this area of research necessitates additional clinical studies and practical insights, especially the development of biomarkers for predicting immune toxicities. In addition, there is a need for heightened education for both clinicians and patients to enhance understanding and awareness.

Comprehensive analyses of the cancer-associated fibroblast subtypes and their score system for prediction of outcomes and immunosuppressive microenvironment in prostate cancer

BackgroundCancer-associated fibroblasts (CAFs) drive cancer progression and treatment failure on one hand, while their tumor-restraining functions are also observed on the other. Recent single cell RNA sequencing (scRNA-seq) analyses demonstrates heterogeneity of CAFs and defines molecular subtypes of CAFs, which help explain their different functions. However, it remains unclear whether these CAF subtypes have the same or different biological/clinical implications in prostate cancer (PCa) or other malignancies.MethodsPCa cells were incubated with supernatant from normal fibroblasts and CAFs to assess their effects on cell behaviors. Sequencing, genomic, and clinical data were collected from TCGA, MSKCC, CPGEA and GEO databases. CAF molecular subtypes and total CAF scores were constructed and grouped into low and high groups based on CAF-specific gene expression. Progression free interval (PFI), clinicopathological features, telomere length, immune cell infiltration, drug treatment and somatic mutations were compared among CAF molecular subtypes and low/high score groups.ResultsThe PCa CAF-derived supernatant promoted PCa cell proliferation and invasion. Based on differentially expressed genes identified by scRNA-seq analyses, we classified CAFs into 6 molecular subtypes in PCa tumors, and each subtype was then categorized into score-high and low groups according to the subtype-specific gene expression level. Such score models in 6 CAF subtypes all predicted PFI. Telomeres were significantly shorter in high-score tumors. The total CAF score from 6 CAF subtypes was also associated with PFI in PCa patients inversely, which was consistent with results from cellular experiments. Immunosuppressive microenvironment occurred more frequently in tumors with a high CAF score, which was characterized by increased CTLA4 expression and indicated better responses to CTLA4 inhibitors. Moreover, this model can also serve as a useful PFI predictor in pan-cancers.ConclusionBy combining scRNA-seq and bulk RNA-seq data analyses, we develop a CAF subtype score system as a prognostic factor for PCa and other cancer types. This model system also helps distinguish different immune-suppressive mechanisms in PCa, suggesting its implications in predicting response to immunotherapy. Thus, the present findings should contribute to personalized PCa intervention.

Exploration of Germline Correlates and Risk of Immune-Related Adverse Events in Advanced Cancer Patients Treated with Immune Checkpoint Inhibitors.

Immune checkpoint inhibitors (ICIs) are increasingly used in the treatment of many tumor types, and durable responses can be observed in select populations. However, patients may exhibit significant immune-related adverse events (irAEs) that may lead to morbidity. There is limited information on whether the presence of specific germline mutations may highlight those at elevated risk of irAEs. We evaluated 117 patients with metastatic solid tumors or hematologic malignancies who underwent genomic analysis through the ongoing Personalized OncoGenomics (POG) program at BC Cancer and received an ICI during their treatment history. Charts were reviewed for irAEs. Whole genome sequencing of a fresh biopsy and matched normal specimens (blood) was performed at the time of POG enrollment. Notably, we found that MHC class I alleles in the HLA-B27 family, which have been previously associated with autoimmune conditions, were associated with grade 3 hepatitis and pneumonitis (q = 0.007) in patients treated with combination PD-1/PD-L1 and CTLA-4 inhibitors, and PD-1 inhibitors in combination with IDO-1 inhibitors. These data highlight that some patients may have a genetic predisposition to developing irAEs.

Emerging Trends In Immunotherapy Management For Pancreatic Cancer: A Literature Review

The prognosis for pancreatic cancer (PC) is poor making it a substantial treatment challenge. A potentially effective treatment option is immunotherapy. To better understand the changing field of immunotherapy in PC management, this analysis will focus on important tactics and their consequences. To find relevant research published between 2019 and 2024, a thorough literature search was carried out using Scopus, Google Scholar, and PubMed. Tumor cells may be targeted and immune responses can be modulated using passive immunotherapeutic techniques such as CAR T-cell therapy and programmed death checkpoint inhibitors. Preclinical and clinical studies show enhanced effectiveness of combination therapy, such as anti-PD-1/anti-PD-L1 plus CTLA-4 inhibitors. Although their efficacy varies, active immunotherapeutic strategies, in particular cancer vaccines, seek to promote immune recognition of tumor antigens. Subpar response rates and treatment-related toxicity are obstacles. Checkpoint inhibitors, CAR T-cell therapy, and cancer vaccines are a few possible therapeutic options in the exciting field of immunotherapy for PC treatment. Nevertheless, further study is required to enhance these strategies and enhance patient.

Abstract 2986: Germline signatures predicting toxicity and response to CTLA4 inhibitors

Abstract Background: Because germline microRNA-based variants predict toxicity risk to anti-PD-1/PDL-1 treatment, and CTLA4 inhibitors are often used in combination with them, we wanted to see if we could also predict toxicity and/or response to anti-CTLA4 therapy. Methods: The study included 69 patients from 3 institutions treated with anti-CTLA4 therapy alone. Patients were 23-89 years old with median age of 62. “Toxicity” was defined as grade 3 or higher toxicity and “Response” as a complete response, partial response, or stable disease. A panel of previously tested variants (n=139), age, sex and cycles of anti-CTLA4 were used to predict endpoints of interest. To balance model dimensionality we pre-screened variants and included them if they were marginally associated with our outcomes of interest via Fisher’s Exact Test or Jonckheere-Terpstra Test at a p-value threshold of < 0.2. We fit Elastic Net (EN), Random Forest (RF), and Boosted Tree (BT) models with up-sampling to predict outcomes of interest. Leave-one-out cross validation (LOOCV) was used to evaluate model performance, and the model with the highest LOOCV AUC (between all EN, RF, and BT) was chosen as the final model and re-fit using all observations. We also performed a stratified gene ontology (GO) analysis to assess biological differences between variants involved in the signatures. Our GO analysis compared gene clusters across groupings with an adjusted p-value cutoff of 0.05 against a universal genomic background. Results: 15 patients had toxicity and 21 patients had a response, with 27/139 and 25/139 variants marginally associated with these endpoints, respectively. Our toxicity model had a LOOCV AUC of 0.811 with the top variables being age, TNNT2_rs3729843, REV3L_rs465646, miR34b_c_rs4938723, and XRCC3_rs861539. Our response model had an LOOCV AUC of 0.716 with top variants being IL2RA_rs2476491, REV3L_rs465646, and IL10RB_rs2834167. Including anti-CTLA4 cycles improved the prediction accuracy for Response with an AUC 0.764. GO analysis revealed 117 pathways enriched in both the Toxicity and Response signatures, which were predominantly involved in pri-miRNA transcriptional regulation. Top GO terms in the Response signature (333 unique) were related to apoptosis and cell death. Top GO terms in the Toxicity signature (425 unique) were related to DNA replication and cellular signaling and response, including the interleukin-35-mediated signaling pathway. Conclusion: We identified germline signatures predicting toxicity and response following anti-CTLA4 therapy. These signatures differed from each other as well as from a previously identified signature of toxicity to anti-PD1/PDL-1 therapy, and GO analysis identified plausible underlying signaling pathways. Our findings continue to validate the potential of these variants to predict meaningful endpoints in cancer therapy, and thus further research in larger and more diverse cohorts is warranted. Citation Format: Joanne B. Weidhaas, Kristen McGreevy, Nicholas Marco, Nora Sundahl, Christopher R. Cabanski, Christine Spencer, Piet Ost, Donatello Telesca. Germline signatures predicting toxicity and response to CTLA4 inhibitors [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 2986.

Abstract 1366: Dual immune checkpoint inhibition enhances the anti-tumor activity of trastuzumab deruxtecan in preclinical models

Abstract Background Trastuzumab deruxtecan (T-DXd) is an antibody-drug conjugate (ADC) composed of an anti-human epidermal growth factor receptor 2 (HER2) antibody, a cleavable tetrapeptide-based linker, and a topoisomerase I inhibitor payload. Recent clinical trials such as BEGONIA have highlighted the potential therapeutic benefit in combining T-DXd with immuno-oncology (IO) agents such as the PD-L1 inhibitor durvalumab. Here, we report preclinical findings demonstrating enhanced anti-tumor activity when T-DXd is combined with dual immune checkpoint inhibition. Methods HER2-expressing human cancer cell lines were treated in vitro with T-DXd and assessed for induction of immunogenic cell death (ICD) markers and expression of NK cell-activating ligands. Human macrophages and T cells were incubated with supernatants from treated cancer cells, and their activation status evaluated via flow cytometry. In vivo, the effects of T-DXd in combination with inhibitors of PD-L1, CTLA-4, and a monovalent bispecific anti-PD-1/TIGIT antibody - a murine surrogate of rilvegostomig - on tumor growth were evaluated in BALB/c mice bearing human HER2-low EMT6 murine mammary tumors. The anti-tumor activity of T-DXd in combination with volrustomig, a monovalent bispecific antibody targeting PD-1 and CTLA-4, was also assessed in hu-CD34+ NSG mice bearing HER2-low Caki-1 renal cell carcinoma tumors. Results In vitro, T-DXd induced ICD via DXd-mediated extracellular release of inflammatory mediators ATP and HMGB1, and cell surface exposure of calreticulin. This was associated with a greater than 2-fold increase in expression of NK cell-activating ligands, and activation of both macrophages and T cells. In immunocompetent mice bearing human HER2 expressing EMT6 tumors, T-DXd treatment drove tumor growth inhibition (53%, P <0.001) that was associated with an increase in tumoral T cells (2.6-fold, P <0.05) and increased CD8+ T cell expression of the immune checkpoints PD-1, TIGIT, and TIM-3. Tumor growth inhibition relative to vehicle-treated mice was enhanced when T-DXd was combined with inhibitors of PD-L1 (108%, P <0.001), PD-L1 plus CTLA-4 (172%, P <0.001), or a bispecific PD-1/TIGIT inhibitor (181%, P <0.001). Pharmacodynamic analysis revealed how the combination of T-DXd and PD-1/TIGIT inhibition increased tumoral NK cells (3.2-fold, P <0.001) and CD8+ T cells (3.8-fold, P <0.001) relative to vehicle. Consistent with findings in syngeneic models, the combination of T-DXd with a bispecific PD-1/CTLA-4 inhibitor also enhanced tumor growth inhibition in a humanised Caki-1 model (121%) compared to T-DXd alone (72%) (P <0.001). Conclusions These data provide insight into the clinical activity observed with T-DXd and provide scientific rationale for combination strategies with novel, bispecific IO agents targeting CTLA-4 or TIGIT in addition to the PD-1/PD-L1 axis. Citation Format: Liam Jenkins, Laura Kazlauskas, Matt Wilson, Scott A. Hammond, Theresa Proia, Jerome Mettetal. Dual immune checkpoint inhibition enhances the anti-tumor activity of trastuzumab deruxtecan in preclinical models [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 1366.

Abstract 3627: Volrustomig a novel PD-1/CTLA-4 bispecific antibody leads to robust increase in peripheral and intra-tumoral pharmacodynamic biomarkers in solid tumors from FTiH study

Abstract Background: Volrustomig (MEDI5752) is designed to fully inhibit PD-1 while preferentially inhibiting CTLA-4 on activated PD-1+ T cells. In a FTiH trial, volrustomig demonstrated meaningful clinical activity in patients with solid tumors (including 1L RCC and NSCLC), acceptable safety profile, sustained PD-1 receptor occupancy, and T cell proliferation at levels greater than seen with clinically tolerable doses of CTLA-4 inhibitors with co-administration of anti-PD(L)-1. We sought to further characterize volrustomig pharmacodynamic effect in the periphery and tumor to demonstrate its biological activity and inform dose optimization. Methods: Pre- and on-treatment blood samples from 86 patients enrolled in dose exploration were collected. Peripheral T cell profiling was assayed by flow cytometry, RNAseq, TCRSeq and serum proteomics. Paired tumor biopsies (N=17), pre- and at week 6 on-treatment, were analyzed for T cell infiltration and function using RNAseq and computational image analysis of immune cell marker density detected by histochemistry and multiplex immune fluorescence assays. Results: Volrustomig achieved dose-dependent peripheral pharmacodynamic changes of biomarkers specific to CTLA-4 blockade at ≥ 225mg. At C1D8 with doses ≥ 500mg, increases in CD4 T cell proliferation (ki67), activation (ICOS), and memory T cells (1516%, 321% and 88%, respectively) were seen, each greater than observed with historical controls of patients treated with tremelimumab 3mg/kg (T3) in combination with durvalumab. Similarly, CD8 proliferation and activation were also higher than T3 at C1D8 with 300% and 161% increase respectively. Additional CD4 and CD8 T cell proliferation and activation was observed at C2D8 supporting the benefit of repeat dosing. Higher peripheral T cell clonal expansion at doses ≥ 500mg was seen and associated with PFS and OS benefit. In patients with paired biopsies, volrustomig achieved robust T cell mediated anti-tumor activity including increases in T cell infiltration (CD8+, 2.3 fold, p=0.04), proliferation (CD8+ Ki67+, 2.1 fold, p=0.04) and, T cell cytotoxicity and activation (GrB+, 2.2 fold, p=0.01; T cell effector and IFNg gene signatures, > 2-fold, p< 0.01). Additionally, volrustomig increased CD8 proliferation in tumor compared to historical control durvalumab alone (>2-fold increase in 70% vs 50% patients). Conclusion: Voltrustomig demonstrated robust peripheral and intra-tumoral T cell activation and proliferation, broadening the pool of antigen experienced T cells, at levels greater than seen with dual checkpoint blockade currently used in the clinic. Citation Format: Ikbel Achour, Michael Kuziora, Florian Song, Steven Eck, Lina Meinecke, Jorge Blando, Jennifer Pearson, Megha Saraiya, Christian Eisen, Felix Segerer, Markus Schick, Nicolas Triltsch, Michael Surace, Gisele Silva Boos, Harald Hessel, Mai Bui, Katrin Pratsch, Alma Andoni, Shelby Denise Gainer, Dan Freeman, Deepa Subramaniam, Ben Tran. Volrustomig a novel PD-1/CTLA-4 bispecific antibody leads to robust increase in peripheral and intra-tumoral pharmacodynamic biomarkers in solid tumors from FTiH study [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 3627.

Synthetic dual co-stimulation increases the potency of HIT and TCR-targeted cell therapies.

Chimeric antigen receptor T cells have dramatically improved the treatment of hematologic malignancies. T cell antigen receptor (TCR)-based cell therapies are yet to achieve comparable outcomes. Importantly, chimeric antigen receptors not only target selected antigens but also reprogram T cell functions through the co-stimulatory pathways that they engage upon antigen recognition. We show here that a fusion receptor comprising the CD80 ectodomain and the 4-1BB cytoplasmic domain, termed 80BB, acts as both a ligand and a receptor to engage the CD28 and 4-1BB pathways, thereby increasing the antitumor potency of human leukocyte antigen-independent TCR (HIT) receptor- or TCR-engineered T cells and tumor-infiltrating lymphocytes. Furthermore, 80BB serves as a switch receptor that provides agonistic 4-1BB co-stimulation upon its ligation by the inhibitory CTLA4 molecule. By combining multiple co-stimulatory features in a single antigen-agnostic synthetic receptor, 80BB is a promising tool to sustain CD3-dependent T cell responses in a wide range of targeted immunotherapies.

Dual Checkpoint Aptamer Immunotherapy: Unveiling Tailored Cancer Treatment Targeting CTLA-4 and NKG2A.

Recent strides in immunotherapy have illuminated the crucial role of CTLA-4 and PD-1/PD-L1 pathways in contemporary oncology, presenting both promises and challenges in response rates and adverse effects. This study employs a computational biology tool (in silico approach) to craft aptamers capable of binding to dual receptors, namely, inhibitory CTLA4 and NKG2A, thereby unleashing both T and NK cells and enhancing CD8+ T and NK cell functions for tumor cell lysis. Computational analysis highlighted AYA22T-R2-13 with HADDOCK scores of -78.2 ± 10.2 (with CTLA4), -60.0 ± 4.2 (with NKG2A), and -77.5 ± 5.6 (with CD94/NKG2A). Confirmation of aptamer binding to targeted proteins was attained via ELISA and flow cytometry methods. In vitro biological functionality was assessed using lactate dehydrogenase (LDH) cytotoxicity assay. Direct and competitive assays using ELISA and flow cytometry demonstrated the selective binding of AYA22T-R2-13 to CTLA4 and NKG2A proteins, as well as to the cell surface receptors of IL-2-stimulated T cells and NK cells. This binding was inhibited in the presence of competition from CTLA4 or NKG2A proteins. Remarkably, the blockade of CTLA4 or NKG2A by AYA22T-R2-13 augmented human CD8 T cell- and NK cell-mediated tumor cell lysis in vitro. Our findings highlight the precise binding specificity of AYA22T-R2-13 for CTLA4-B7-1/B7-2 (CD80/CD86) or CD94/NKG2A-HLA-E interactions, positioning it as a valuable tool for immune checkpoint blockade aptamer research in murine tumor models. These in vitro studies establish a promising foundation for further enhancing binding capacity and establishing efficacy and safety in animal models. Consequently, our results underscore the potential of AYA22T-R2-13 in cancer immunotherapy, offering high specificity, low toxicity, and the potential for cost-effective production.

Identification of immune cell-related prognostic genes characterized by a distinct microenvironment in hepatocellular carcinoma.

The development and progression of hepatocellular carcinoma (HCC) have been reported to be associated with immune-related genes and the tumor microenvironment. Nevertheless, there are not enough prognostic biomarkers and models available for clinical use. Based on seven prognostic genes, this study calculated overall survival in patients with HCC using a prognostic survival model and revealed the immune status of the tumor microenvironment (TME). To develop a novel immune cell-related prognostic model of HCC and depict the basic profile of the immune response in HCC. We obtained clinical information and gene expression data of HCC from The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) datasets. TCGA and ICGC datasets were used for screening prognostic genes along with developing and validating a seven-gene prognostic survival model by weighted gene coexpression network analysis and least absolute shrinkage and selection operator regression with Cox regression. The relative analysis of tumor mutation burden (TMB), TME cell infiltration, immune checkpoints, immune therapy, and functional pathways was also performed based on prognostic genes. Seven prognostic genes were identified for signature construction. Survival receiver operating characteristic curve analysis showed the good performance of survival prediction. TMB could be regarded as an independent factor in HCC survival prediction. There was a significant difference in stromal score, immune score, and estimate score between the high-risk and low-risk groups stratified based on the risk score derived from the seven-gene prognostic model. Several immune checkpoints, including VTCN1 and TNFSF9, were found to be associated with the seven prognostic genes and risk score. Different combinations of checkpoint blockade targeting inhibitory CTLA4 and PD1 receptors and potential chemotherapy drugs hold great promise for specific HCC therapies. Potential pathways, such as cell cycle regulation and metabolism of some amino acids, were also identified and analyzed. The novel seven-gene (CYTH3, ENG, HTRA3, PDZD4, SAMD14, PGF, and PLN) prognostic model showed high predictive efficiency. The TMB analysis based on the seven genes could depict the basic profile of the immune response in HCC, which might be worthy of clinical application.

Immunotherapy in Cancer Treatment: Harnessing the Power of the Immune System

Immunotherapy has revolutionized cancer treatment by leveraging the body's immune system to fight against cancer cells. This article provides an overview of immunotherapy, focusing on its different modalities, mechanisms of action, clinical applications, and the management of immune-related adverse events. Immune checkpoint inhibitors, such as PD-1 and CTLA-4 inhibitors, are prominent modalities that enhance the immune response by blocking regulatory proteins. Additionally, CAR-T therapy genetically modifies a patient's T cells to target specific proteins on cancer cells, leading to precise cancer cell elimination. Immunotherapy has demonstrated remarkable success in certain malignancies and offers new hope for patients battling cancer.