bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting CD47 in lymphoid malignancies with CAR T cells.
Authors: Boardman Alexander Paul , James Scott , Van Den Brink Marcel R. M. 
Published: June 2023

2513 Background: Despite major therapeutic advances of chimeric antigen receptor (CAR) T cells directed against CD19 in relapsed and refractory non-Hodgkin lymphoma (NHL), two thirds of patients experience relapse within the first two years. Nearly one third of these relapses are associated with CD19 antigen-loss escape thus leveraging alternate CAR targets may improve outcomes. CD47, an anti-phagocytic “don’t eat me” receptor known to be overexpressed in NHL, is an ideal CAR target as it is potentially indispensable for tumor progression. Methods: CD47-CAR binding domains were engineered either as a camelid single variable heavy chain nanobody (CD47-VHH-CAR) or as the CD47 binding domain of cognate receptor SIRPα (SIRPα-CAR), each combined with a CD28 costimulatory domain and a CD3ζ activation domain with two mutated ITAMs (1XX). In vitro co-culture assays were performed with lymphoid mouse tumor cell lines BM185 and A20. For in vivo experiments, syngeneic BALB/c mice were sublethally irradiated for lymphodepletion then injected intravenously with 1x105 luciferase+ BM185 cells. Two days later, mice were injected with 2x106 CAR T cells, and bioluminescence imaging was performed to monitor tumor progression. For bio-localization experiments, CAR T cells were co-transduced with a separate vector encoding luciferase. Results: CD47-CAR T cell expansion in vitro was hindered due to fratricide, which can be rescued by CD47 CRISPR knockout. Both CD47-VHH-CAR and SIRPa-CAR T cells exhibited potent and specific in vitro lysis of BM185 and A20 cells. CD47-VHH-CAR T cells showed transient anti-leukemia activity, however, survival was not significantly prolonged. T cell bio-localization analysis revealed that, compared to CD19-CAR T cells, CD47-VHH-CAR T cells exhibited prolonged retention in the thorax with minimal bone marrow trafficking, suggesting CD47 antigen-specific pulmonary sequestration, and inferior expansion. SIRPα-CAR T cells engineered to secrete pro-inflammatory cytokine IL-18 significantly improved CAR T cell expansion, resulting in bone marrow trafficking and clearance of BM185 tumor. However, recipients of IL-18-secreting SIRPa-CAR T cells experienced lethal CAR T cell-related toxicity. To improve the safety of CD47-CAR T cells, we engineered an NFAT-responsive vector expressing the CD47-CAR modified with E. coli-derived dihydrofolate reductase drug destabilization domain to limit CD47-CAR expression to T cells exposed to the stabilizing antibiotic trimethoprim and receiving a licensing NFAT signal from a CD19-CAR. Conclusions: In vivo activity of CD47-targeting CAR T cells is limited by thoracic retention and limited expansion, which can be partially overcome by overexpression of IL-18, though enhanced toxicity precludes survival prolongation. Our findings suggest that strategies to spatiotemporally restrict CAR expression or activation will be necessary to achieve safe and efficacious CD47-CAR T cell activity.
Effect of a novel expansion process on tumor-infiltrating lymphocyte (TIL) polyfunctionality, cytotoxicity, and expansion, while preserving cells in a less differentiated and more stem-like phenotype.
Authors: Yuhas Andrew , Dean Joe , Erickson Tim , Machin Marcus , Alkhouli Mohammed , Yglesias Joe , Cubas Rafael 
Published: June 2023

2542 Background: Adoptive cell therapy using autologous TIL has shown durable responses in patients with metastatic melanoma and some epithelial tumors (Chesney JITC 2022; Schoenfeld SITC 2021; O’Malley SITC 2021). Data suggest that maintaining a higher proportion of less differentiated and more stem-like TIL can associate with persistence and response in patients with metastatic melanoma (Krishna Science 2020; Rosenberg Clin Cancer Res 2011). Here, we describe a novel TIL expansion process that increases TIL expansion and preserves cells in a less-differentiated and more stem-like phenotype with enhanced functional output. Methods: Tumors of various histologies, including lung, breast, and renal cancer, were fragmented and expanded using either a standard process or a newly developed process that uses a different combination of cytokines during the pre-rapid expansion protocol (pre-REP) stage and another combination of cytokines and a pathway inhibitor during the REP stage to control T-cell activation and differentiation. The expansion potential, viability, and phenotypic and functional attributes of the final TIL products were evaluated by a variety of assays. Results: When compared with standard TIL expansion, the novel process increased yield and viability while preserving TIL in a less differentiated and more functional state, as evidenced by increased expression of the memory-associated markers CD27, CD28, CD62L, and IL-7R, with reduced expression of the activation markers CD38, CD39, and CD69, and lower levels of the inhibitory markers LAG3, TIM3, TIGIT, and TOX. Importantly, the novel process led to a pronounced increase in the tumor-homing marker CXCR3 as well as TIL polyfunctionality as evidenced by increased co-expression of IFNγ, TNFα, and IL-2 while showing an enrichment in genes associated with stem-like cells and a reduction in exhaustion-associated genes. Pseudotime trajectory analysis also demonstrated that the new expansion process maintained TIL in a less differentiated and more stem-like state. Together, these phenotypic and functional characteristics translated into increased cytotoxicity even after repeated stimulation. Conclusions: Our novel TIL expansion process improves multiple metrics that correlate with both TIL persistence and response, including enhanced polyfunctionality, reduced inhibitory receptor expression and a less differentiated and more stem-like phenotype, while increasing yield. These effects may translate into a more vigorous and less differentiated TIL infusion product with improved cytotoxicity and persistence.
Phase I results of human endogenous retrovirus type-E (HERV-E) TCR transduced T-cells in patients (pts) with metastatic clear cell renal cell carcinoma (mccRCC).
Authors: Nadal Rosa , Barisic Stefan , Scurti Gina M. , Cherkasova Elena , Chen Long , Wood Kristen , Highfill Steven L. , Wells Brian , Aue Georg , Shalabi Reem , Hughes Thomas E. , Tian Xin , Xu Yifei , Malayeri Ashkan A. , Bagheri Mohammadhadi H. , Reger Robert N. , Nishimura Michael I. , Childs Richard W. 
Published: June 2023

2549 Background: The CT-RCC HERV-E is expressed in the majority of ccRCC tumors with no expression in normal tissues. T-cells transduced to express a HERV-E TCR are cytotoxic to ccRCC & mediate tumor regression of human ccRCC in a murine model. We developed a method to manufacture HERV-E TCR transduced T-cells (HERV-E TCR) for clinical development. Methods: We conducted a phase I dose-escalation trial (DL1:1x106, DL2:5x106, DL3:1x107 & DL4:5x107cells/kg) to evaluate the safety of adoptively infused HERV-E TCR in mccRCC pts. HERV-E TCR contain T-cells engineered to express an HLA-A*11 restricted HERV-E TCR and a truncated CD34 cassette for in vivo monitoring. Eligible HLA-A*11 pts had mccRCC, ECOG 0-1 & prior antiangiogenic & checkpoint inhibitors therapy unless contraindicated/unavailable. Cyclophosphamide & fludarabine were given followed by HERV-E TCR infusion & IL-2 administration. Results: 14 HLA-A*11+ pts (median age 56) were treated including 3 pts in each DLs 1-3 & 5 pts in DL4. 86% had ≥ 3 prior treatment lines (range 1-7): 36% had high-dose IL2, 57% Ipilimumab-Nivolumab & 50% ≥ 3 lines of anti-VEGFR therapy. Bone, liver & brain metastases were present in 71%, 43%, and 14% of pts. All HERV-E TCR products met sterility, purity, and potency criteria for release and infusion (Table). No HERV-E TCR dose-limiting toxicities or treatment-related deaths occurred. Pts received a mean: 12 IL-2 doses (IQR 11-14). 57% pts had G3-4 febrile neutropenia & 7% G3-4 capillary leak syndrome. 1 pt experienced G2 skin rash possibly related to HERV-E TCR. Best response included 7% pts with partial response & 29% with stable disease ≥ 8 weeks. The mPFS: 62 days (IQR 31,90). At DL4, TCR transgene expression was detectable in PBMCs at a mean 6.3% (range (r) 0.02-25), 12.3% (r 0.01-61.2), 3.45% (r 0.00-13.7) & 2.89% (r 0.00-11.5) on days 4, 7, 14, & 21. HERV-E TCR were detected in the malignant pleural effusion of 1 pt. Conclusions: This is the first trial evaluating TCR-engineered T-cells targeting a human endogenous retrovirus in ccRCC. The manufacturing method utilized produced large numbers of highly purified CD8+ HERV-E reactive T-cells that were not associated with any dose limiting toxicities when given at doses up to 5x107 cells/kg. For pts in DL4, HERV-E TCR were observed to proliferate in vivo, traffic to a metastatic site, and induce tumor regression in one mccRCC pts. Clinical trial information: NCT03354390 . [Table: see text]
Preclinical testing of FOLR1-CAR T cells against osteosarcoma (OS).
Authors: Choe Michelle , Kirkey Danielle , Ries Rhonda E , Hylkema Tiffany , Menashe Sarah J , Davis Lara E. , Wagner Michael J , Cranmer Lee D. , Malhotra Sanjay , Gorlick Richard Greg , Meshinchi Soheil , Loggers Elizabeth Trice 
Published: June 2023

2555 Background: Survival rates for OS are poor. Prior literature suggests folate receptor-a (FOLR1) overexpression in OS, making it a target for CAR T development. Methods: OS transcriptome data from TCGA, PCGP and NCI PIVOT programs were interrogated for FOLR1 gene expression. A 2nd generation FOLR1-CAR T cell was created by fusing the ScFv from farletuzumab (anti-FOLR1 monoclonal antibody) with a 4-1BB costimulatory and CD3z cytotoxicity domain. Non-transduced (NT) T cells served as controls. In vitro functional analysis was performed against U2OS cell line using flow cytometry (FC) and Incucyte. U2OS xenograft in NSG mice was generated via intra-femoral inoculation for local tumor and via tail vein (TV) tumor injection for pulmonary disease. Tumor engraftment was confirmed by in vivo imaging system (IVIS). Disease progression was monitored by IVIS and CT imaging. 5 million T cells were administered via TV. Circulating human T cells (HTC) collected from retro-orbital peripheral blood (PB) were detected via FC. Results: FOLR1 transcript was expressed in >75% of OS in TCGA, PCGP and PIVOT. In vitro evaluation of FOLR1-CAR T demonstrated robust antitumor activity (E:T ratio 1:1) with cytotoxicity of 76.5% and 95.5% at 6 and 24 hrs. Disease progression was observed in NT T cell treated mice, with an average fold increase in IVIS radiance of 35.1 in local (n=2) and 142.8 in pulmonary disease (n=3) over 11 weeks (wks) versus complete resolution of local (n=2) and pulmonary disease (n=3) in FOLR1-CAR T treated mice (see table). NT T treated mice with local disease demonstrated significant bone destruction; no local tumors were found in FOLR1-CAR T treated mice. HTC (CD45+/CD3+) expansion was detected in PB one wk post-treatment (PT); in mice with metastasis 0.190% and 0.136% of live circulating cells were human T cells in FOLR1-CAR T and NT T treated mice, respectively. Improved overall survival was observed in FOLR1-CAR T treated mice (>100 vs 87.5 days). Conclusions: FOLR1-CAR T cells appear to have in vivo activity against U2OS cell line and support further testing of safety and feasibility in an early phase clinical trial for relapsed/refractory OS. [Table: see text]
A novel platform to enrich for tumor-specific T cells in non–small-cell lung cancer.
Authors: Hachey Stephanie J , Forsythe Alexander G , Mahtabifard Ali , Keshava Hari B. , Hughes Christopher C.W. 
Published: June 2023

2556 Background: To induce robust cellular immunity against non-small cell lung cancer (NSCLC), the leading cause of cancer-related death in the US, we have developed an innovative cell-based therapeutic platform technology to generate polyclonal, neoantigen-reactive T cells in vitro. Methods: Surgically resected NSCLC tissue and peripheral matched blood were collected from 17 patients under an IRB-approved protocol. From 3 of the patient-derived NSCLC specimens, methods were tested to enhance immunogenicity and stimulate neoantigen-specific immune responses. We then use a novel, human vascularized microtumor (VMT) platform to assess the sensitivity of tumor cells to T-cell mediated attack at the individual patient level. Results: The therapeutic platform induced a significant increase in T cell proliferation, interferon-gamma secretion, and blast phenotype of lymphocyte populations compared to T cells primed by standard methods. Interestingly, for one patient, immunosuppression of the T cell response with concomitant increase in CD96, an immune checkpoint receptor implicated in lung cancer progression, was observed in the tumor. Importantly, using sequencing data from one patient, an analytical and machine learning pipeline detected putative neoantigens using our therapeutic platform technology and not with standard methods. Conclusions: The therapeutic platform can be used to directly generate cell-based immunotherapies and to facilitate neoantigen discovery and development of targeted cancer immunotherapies. Our goal is to advance novel approaches to effectively mobilize anti-tumor immunity and overcome treatment resistance in NSCLC.
A subset of neutrophils as a predictive biomarker for immunotherapy response in patients with non–small-cell lung cancer and melanoma.
Authors: Shaked Yuval , Benguigui Madeleine , Halaban Ruth , Bacchiocchi Antonella , Kamer Iris , Bar Jair , Lotem Michal , Shen-Orr Shai , Sznol Mario , Cooper Tim J 
Published: June 2023

2557 Background: Despite the remarkable success of immunotherapy, only ~20-40% of cancer patients display an extended durable response. Pre-existing biomarkers for immunotherapy outcome are significantly limited in their predictive power, e.g., PDL-1 expression by tumor cells (AUC ~ 0.6-0.75). This requires additional efforts to search for new approaches to discover new biomarkers in immune-oncology. While mice are the most widely used and cost-effective model to study human disease, translating preclinical biomarkers into clinical practice faces significant obstacles – in part due to the lack of diverse or appropriate models. In this study we have taken a holistic approach to search for new biomarkers for immunotherapy outcomes, with the aim to improve translatability of the findings and their validity in cancer patients. Methods: Several pre-clinical models were used, each capturing one possible mechanistic aspect of immunotherapy response, such as tumor- and host-dependency, in order to reflect the variability seen in human cancers. These models have been analyzed for myeloid cell composition within tumors and peripheral blood using high throughput techniques such as single-cell RNA sequencing and mass cytometry, followed by rigorous bioinformatic tools. Peripheral blood mononuclear cells (PBMCs) were obtained from non-small cell lung cancer (NSCLC, n = 34) and melanoma (n = 38) patients prior to immune-checkpoint inhibitor backbone therapy as monotherapy or in combination with chemotherapy. These results were validated on additional human bulk mRNA datasets (n = 417). Results: Using the preclinical models, we identified interferon-stimulated, Ly6Ehi neutrophils as a pre-treatment, blood-borne biomarker for anti-PD1 response. These results were validated in PBMCs of advanced metastatic NSCLC and malignant melanoma patients predominately treated with immune checkpoint inhibitor-based therapy, with a high degree of accuracy (AUC ~ 0.9). These results were also validated in independent cohorts of immunotherapy-treated patients with other cancer types obtained from bulk mRNA datasets including renal cell carcinoma (n = 109), urothelial carcinoma (n = 228) and additional melanoma patients (n = 80). Functional studies revealed that Ly6Ehi neutrophils sensitize otherwise resistant tumors to anti-PD1 therapy, in part by directly activating cytotoxic T cells and contributing to tumor cell killing, while operate upstream of T cells in the immunotherapy response. Conclusions: Our study demonstrates a pragmatic approach to search for new, clinically relevant and functionally active cellular biomarkers for immunotherapy outcomes in humans. This study paves the way for a conceptual framework to advance the field of immuno-oncology by utilizing this approach for successful clinical trials.
C-reactive protein as a response biomarker to immune checkpoint blockade: A meta-analysis.
Authors: Karol Alexander B , Fujiwara Yu , D'Ovidio Tyler J , Baldwin Elena , Joshi Himanshu , Doroshow Deborah Blythe , Galsky Matt D. 
Published: June 2023

2559 Background: Tumor-promoting inflammation is a hallmark of cancer pathogenesis and is associated with poor outcomes and treatment resistance. C-reactive protein (CRP) is a routinely measured acute phase reactant whose synthesis is stimulated by cytokines and may thereby represent a surrogate for tumor promoting inflammation. Pre-treatment CRP has been associated with resistance to immune checkpoint blockade (ICB) in several studies. However, whether post-treatment changes in CRP correlate with ICB outcomes has not been systematically examined. Methods: We performed a systematic review to identify cohort studies or clinical trials in solid tumor patients receiving ICB therapy with CRP response comparator pairs (high/low) available. We included studies that had hazard ratio (HR) of overall survival (OS), progression-free survival (PFS), or odds ratio (OR) of objective response rate (ORR) available. We performed a meta-analysis using a random-effect model to evaluate if post-treatment changes in CRP are associated with ORR, PFS, or OS. Subgroup analysis for primary cancer type was performed. Heterogeneity was assessed using the I-squared statistic. Results: We screened 691 eligible studies; 19 met inclusion criteria and 2,101 patients were included. Patients experiencing post-ICB declines in CRP had improved ORR HR=4.67 [95% CI] [2.81-7.78] (p<0.0001, I2=42%), PFS HR=2.47 [1.98-3.09] (p<0.0001, I2=18%), and OS HR=2.28 [1.68-3.08] (p<0.0001, I2=56%). Heterogeneity among subgroups was low in ORR (I2=14.6%) and PFS analysis ( I2=0%), and moderate in OS analysis (I2=64.2%). Conclusions: In patients receiving ICB therapy, post-treatment declines in CRP were associated with improved ORR, PFS, and OS across multiple histologies. These findings support the integration of on-treatment CRP decline as a clinically relevant response biomarker for novel therapies directed at modulating tumor-promoting inflammation. A meta-regression analysis will help determine optimal post-treatment CRP cutpoints. Pooled OR of ORR [95%CI] and HR [95%CI] of PFS and OS with subgroup analysis results and heterogeneity assessment. [Table: see text]
Time-dependent efficacy of immune checkpoint inhibitors in the treatment of metastatic cancers: A meta-analysis.
Authors: Landre Thierry , Karaboue Abdoulaye , Buchwald Zachary S , Innominato Pasquale , Qian David C , Assie Jean-baptiste , Chouaid Christos , Levi Francis , Duchemann Boris 
Published: June 2023

2562 Background: Immune checkpoint inhibitors (ICIs) have been established as the standard of care for numerous advanced malignancies. Emerging evidence suggests that the time-of-day of administering these immunotherapies may impact patient outcomes. We sought to determine the consistency in the impact of ICI time-of-day infusion on survival of patients with metastatic cancer. Methods: We performed a meta-analysis of published studies that have reported on ICI time-of-day infusion and efficacy in patients with advanced solid tumors. Overall survival (OS) and progression free survival (PFS) were compared between patients who had received ICIs before and after the daily time cut-off of each study. Hazard ratios (HRs) with their 95% confidence interval (CI) were collected from the studies and pooled. Results: This meta-analysis identified 7 studies of 1019 patients (PS 0, 43%; PS 1, 39%; males, 65%; females, 35%) who had stage IV non-small cell lung cancer (NSCLC), urothelial cancer, renal cell carcinoma, or melanoma, and were treated with mostly anti-PD-1 (nivolumab or pembrolizumab), anti-PD-L1 (atezolizumab), and anti-CTLA-4 (ipilimumab) agents. The time cut-offs selected to stratify patterns of ICI infusion were 16:00 or 16:30 in 5 studies (based on differences in adaptive immune response reported in previous vaccination studies) and 13:00 (based on median time of ICI infusions) in 2 studies. The pooled analysis of OS results showed a statistically significant benefit in favor of patients receiving infusions in the morning or in the early part of the day with a HR: 0.49, [95% CI: 0.36-0.69] (p < 0.0001). A similar benefit was observed for PFS with a HR: 0.56, [0.44-0.72] (p < 0.00001). Conclusions: Patients with advanced cancers may benefit from earlier time-of-day ICI infusions. Our findings are consistent with established circadian mechanisms that control immune cell functions and trafficking over a 24-h period. Prospective randomized trials are warranted to establish ICI timing recommendations for optimizing treatment efficacy in patients with different cancer types and at minimal cost.
Phase 1 clinical trial results for NG-350A, a novel transgene-armed and tumor-selective vector: Differential effects of intravenous (IV) versus intratumoral (IT) dosing on immune pharmacodynamics (PD).
Authors: Patel Minesh , Cox Chris , Krige David , Walker Lyle , Evilevitch Vladimir , Carter Jo , Thomas Matthew , Bucktrout Samantha , Champion Brian , Lillie Thomas , Rosen Lee S. , Naing Aung 
Published: June 2023

2572 Background: Tumor-selective therapeutic viral vectors are often limited to IT delivery. NG-350A, a blood stable adenoviral vector, selectively replicates in tumors and expresses an agonist anti-CD40 payload. We assessed the impact of IV vs IT delivery during the phase 1 FORTITUDE study (NCT03852511). Methods: NG-350A was dose-escalated in separate IT or IV cohorts of patients (pts) with metastatic/advanced epithelial tumors. Pts received either 1 (Day [D] 1) or 4 IT injections (D1, 8, 15, 22) of up to 1x1012 viral particles (vp) or 3 IV infusions completed in <9 days (doses of 1x1011 to 6x1012 vp). Serum samples were taken immediately after dosing (pharmacokinetics [PK]) and during follow-up (PD). Baseline and on-treatment (D15+) tumor biopsies were taken. Results: In total, 25 pts received NG-350A monotherapy, 9 by IT and 16 by IV dosing. Both IV and IT dosing elicited similar anti-drug (virus) antibody responses and limited viral shedding. Peak vp Cmax was variable with IT dosing but dose-related with IV dosing. Vector DNA was detected by qPCR in tumor biopsies after higher IV dose levels (including up to ~7 weeks post dosing) and IT injection. Dose-dependent, sustained and specific increases in inflammatory cytokines related to the expected effects and timing of the anti-CD40 transgene production (IL-12, IL-17A, IL-2, IFNγ) were detected in serum of pts treated at higher IV doses. By contrast, limited evidence of such PD effects was seen with IT dosing. Transgene was not detectable in core needle biopsies; however, a novel serum biomarker assay detected transgene mRNA at D8 or D15 in five pts who received higher IV dose levels. Importantly, both IV and IT dosed NG–350A was well-tolerated, with no treatment-related SAEs occurring in >1 pt and no DLTs at the highest dose levels tested. No evidence of anti-CD40 transgene-related toxicity was seen with IV or IT dosing, with few hepatic AEs and no Grade ≥3 CRS. Follow-up for imaging was limited; however, stable disease was observed in 3/6 pts at IV Dose Level 3. Conclusions: This analysis provides the first clear evidence that an IV-dosed tumor-selective viral vector can drive expression of immuno-stimulatory payloads with functional PD consequences in advanced cancers. The improved PK and PD profile of IV vs IT dosed NG-350A also highlights the benefits of systemic delivery for such vectors. Dose-escalation of NG-350A + pembrolizumab to identify a dose for efficacy assessments continues in the FORTIFY study (NCT05165433). Clinical trial information: NCT03852511 . [Table: see text]