bluuebunny/crossref_metadata_2025_split huggingface dataset:
Peri Cruiser CAR-T: An innovative platform to reduce on-target off-tumor toxicity of CAR-T therapy.
Authors: zhang shuangshuang , Hao Ruidong , Wang Hongye , Yi Qiao yong , Yantao Li , Zhong Yunpeng , Sun Minmin 
Published: June 2023

2539 Background: On-target off-tumor toxicity is a significant concern in Chimeric Antigen Receptor (CAR)-T therapies, especially for solid tumors, leading to adverse effects on normal tissues and impacting on treatment efficacy. To address this challenge, a modified form of CAR-T cells named Peri Cruiser CAR-T has been developed by using miRNA technology to knock down three adhesion genes that mediate T cell transendothelial migration to normal tissues, aiming to block the extravasation of CAR-T cells so that the on-target off-tumor toxicity could be prevented without compromising anti-tumor efficacy. In this study, we provided a proof of concept by using the epithelial cell adhesion molecule (EpCAM) as a targeting tumor-associated antigen (TAA). Methods: An miRNA cassette was incorporated into the vectors of anti-human or mouse EpCAM CARs. The knockdown efficiency, endothelial adhesion efficiency, effector functions and memory phenotypes in vitro were evaluated. To determine the toxicity in vivo, the anti-mouse EpCAM CAR-T cells were administered to non-tumor bearing NSG mice, and the body weight and T cell infiltration in normal tissues was evaluated. Anti-tumor efficacy was also evaluated in NSG mice with subcutaneous and metastatic tumor models by anti-human EpCAM CAR-T cells. Results: Our Peri Cruiser CAR-T platform was capable to effectively downregulate the target genes by 10 folds. The knockdown is stable after extended culture and antigen engagement in vitro. This knockdown of targeted genes significantly decreased T cell adhesion to endothelial cells and corresponding ligands, but did not affect T cell activation or effector functions such as cytotoxicity and IFN gamma release. Surprisingly, lowering the levels of these adhesion molecules improved the memory phenotypes of CAR-T cells as indicated by increased portion of CCR7+CD45RO+ cells. In vivo studies showed that conventional EpCAM CAR-T-injected mice experienced severe weight loss and all had to be sacrificed before day 8, the IHC staining of CD3 revealing massive infiltration of T cells in lung, liver, and kidney. However, the incorporation of the miRNA cassette prevented T cell infiltration in normal tissues, reversed the toxicity, and ensured the survival of all mice. The anti-tumor efficacy of these T cells was evaluated in vivo and found to decrease metastasis as effectively as the conventional CAR-T cells in a metastasis model of HCT116-Luc, although there was a slightly but not significantly decreased efficacy in a subcutaneous model of HCT116. Conclusions: The Peri Cruiser CAR-T platform can significantly improve the safety of CAR-T cells without compromising their efficacy to eliminate tumor cells in primary site and metastasis. This technology opens another avenue to safely target a variety of other TAA by CAR-T even when there is the potential of on-target off-tumor toxicity in normal tissues for those antigens.
Selective recognition of over-expressed self-antigens in solid tumors using calibrated CAR-T therapy.
Authors: Haerizadeh Farzad , Jiang Helena , Cooper Laurence 
Published: June 2023

2540 Background: CAR-T has shown promise in patients with a subset of solid tumors. Future development depends on identifying CAR-T that differentiate between normal and malignant cells. The PrismCore platform develops CAR-T targeting self-antigens using three principles: 1) identification of targets uniformly expressed on solid tumors, 2) over-expression on the surface of invasive cancers compared to healthy cells, and 3) elevated levels linked to tumor aggressiveness. PrismCore produces CAR-T for treating multiple solid tumor types by calibrating T-cell activity against self-antigens only when present at elevated densities. Methods: PrismCore generates CAR-T by combining computational biology with empirical observations to target over-expressed self-antigens. Proto-libraries of CARs are created using synthetic single-domain antibodies and structural/signaling components. Bio-screening is performed using combinatorial libraries generated through computational simulation. Lead CAR-T are identified through iterative in vitro and in vivo testing and evaluated for safety in non-human primates (NHP). Results: PrismCore platform has identified engineered T cells against several self-antigens, with the first CAR-T, termed B4t2-001, advanced to clinical testing against the target BT-001. Libraries (1E10 to 1E11) of camelid antibodies were designed, synthesized, and assessed. Selected antibody variants subsets were combinatorially assembled into CAR-Ts and rigorously screened against gradient of BT-001. Six candidates were advanced to further testing in mice to evaluate anti-tumor effect and sustained in vivo biological activity. B4t2-001 was evaluated in NHP that express basal levels of BT-001. No toxicity observed despite measurable circulating CAR-T and in vitro activity to NHP BT-001 when at high density. The turnaround from target selection to B4t2-001 for clinical evaluation was six months. Conclusions: We report pre-clinical data for B4t2-001 that safely and effectively targets BT-001 when overexpressed on tumors. This is a novel target for T-cell therapy and is at elevated levels on multiple different types of solid tumors. BT-001 participates in pathobiology, tumor invasion as well as metastasis, and level of expression inversely correlates with overall survival. B4t2-001 demonstrated target density-dependent response, potency, durability of effect and safety in preclinical models and is undergoing phase I clinical evaluation (NCT05621486) in patients with different types of solid tumors. [Table: see text]
Effect of the killing style of individual CAR T cells on the clinicopathological behavior of therapeutic products in the clinic.
Authors: Gad Ahmed Z , Morris Jessica S , Godret-Miertschin Lea , Montalvo Melisa J , Berger Harrison , Brock Rebecca M , Fousek Kristen , Srinivasan Madhuwanti , Saeedi Arash , Levental Kandice , Mamonkin Maksim , Carisey Alexandre , Varadarajan Navin , Hegde Meenakshi , Joseph Sujith , Levental Ilya , Mukherjee Malini , Ahmed Nabil 
Published: June 2023

2548 Background: CAR T cell products that signal through CD28 and 41BB are highly effective against B lineage leukemia, despite exhibiting distinct efficacy and toxicity profiles in patients. Specifically, CD28 has been linked to higher upfront lytic activity and toxicity, while 41BB induces longer CAR T cell persistence and more durable progression free survival. We reasoned that the killing style of individual CAR T cells can explain the clinicopathological behavior of CAR T cell products in the clinic. Methods: We studied the cellular kinetics of CD28 and 41BB signaling CD19 CAR T cell products approved in the clinic, their single cell killing behavior in high throughput, the molecular composition of the lipid rafts inside the CAR immune synapse (CARIS) they form with their target cells, and their CARIS mechanical properties, then corroborated these findings in one kinetic model to explain their distinct clinicopathological behavior in the clinic. Results: We discovered that tumor encounter polarizes CD28 CAR T cell products to a predominance of more differentiated CD8 effectors that rely on short-lived highly lethal synapses with tumor cells. CD28 CAR T cell mediated synapses are characterized by fast mechanical buildup and resolution and brisk shuttling of CAR and other associated molecules into the CARIS and rapid polarization of the killing machinery in a manner that supports a mastery of serial killing mediated by perforin and granzymes. In contrast, 41BB responds to tumor stimulation by a Th2 -polarizing cytokine milieu that robustly expands a balanced CD4/CD8 population of effectors preserving a central memory phenotype. 41BB CAR T cells engage in lengthier synapses characterized by slower accumulation of CAR molecules in a mechanically tonic CARIS and rather collaborative yet lengthy Fas ligand-mediated killing process. Conclusions: We show here that CAR T cell signaling domains mediate distinct killing dynamics that can explain their behavior in patients. By understanding and potentially modulating these dynamics, the CAR T cell functionality and toxicity can be rationally optimized.
Preclinical evaluation of targeting PRAME with TCR mimic CAR T cells in AML.
Authors: Kirkey Danielle , Robinson Leila , Hylkema Tiffany , Loeb Anisha , Castro Sommer , Tang Thao , Ries Rhonda E , McKay Cyd , Root Christina , Pardo Laura , Loeb Keith , Le Quy , Meshinchi Soheil 
Published: June 2023

2553 Background: Chimeric antigen receptor (CAR) T cell therapy has revolutionized cancer treatment, but has had limited success against AML in part due to overlap of cell surface antigens expressed in AML and normal hematopoietic cells. To identify AML-restricted targets, we interrogated the transcriptome from over 3000 AML patients and found PRAME (Preferentially Expressed Antigen in Melanoma), an intracellular protein, to be a highly expressed AML-restricted target. Using a novel approach to target intracellular antigens, we developed a PRAME CAR T cell using a TCR mimic (mTCR) antibody, which recognizes the PRAME peptide/HLA-A2 complex on the tumor cell surface. To conduct final IND-enabling studies, we generated an in vivo patient derived xenograft (PDX) model to treat with PRAME mTCRCAR T cells. Here we demonstrate the in vivo activity of PRAME mTCRCAR T cells against a PDX AML model. Methods: We used the VL and VH sequences from the PRAME TCR mimic antibody (Pr20) to construct the single chain fragment variable domain into the 41-BB/CD3ζ CAR vector. The PDX model was derived from a PRAME+/HLA-A2+ pediatric patient with AML. PDX cells were transduced with ffluciferase for noninvasive bioluminescent IVIS imaging to monitor leukemic progression. PDX leukemia-bearing mice were treated with unmodified T-cells or PRAME mTCRCAR T cells at 5x106 cells (1:1 CD4:CD8) per mouse 1 week after leukemia injection. Leukemia burden was measured by IVIS imaging and peripheral blood analysis. Results: Treatment with PRAME mTCRCAR T cells led to eradication of leukemia with all mice alive >100 days post-treatment, while control mice (unmodified T cells) had disease progression at Day 50 (p=0.001). Average leukemia burden in the control cohort was 3.5% at week 6 and 41.75% at week 11, while no leukemia was detected following CAR T cell treatment. In addition, the control arm had a marked expansion of leukemia with a 7.75 and 429.6 fold increase in radiance by IVIS at 6 and 11 weeks post-treatment. No significant increase in radiance was seen in the PRAME mTCRCAR T cell treated group. Human T cells (CD45+/CD3+) were detectable in the peripheral blood at Day 7 and 14 post-treatment in both the unmodified and CAR T cell treated groups. Conclusions: We demonstrate the therapeutic potential of targeting PRAME with mTCRCAR T cells in AML. We show potent efficacy with eradication of leukemia in PDX-bearing mice following treatment with PRAME mTCRCAR T cells resulting in prolonged survival. These results provide a novel approach to target PRAME with CAR T cells and provide compelling data to evaluate PRAME mTCRCAR T cells for use in clinical trials against AML. [Table: see text]
The exploratory case-control study on prediction of immune checkpoint inhibitor-induced thyroiditis by human leukocyte antigen types.
Authors: Natori Yutaka , Sasaki Eisaku , Suga Junko , Kimura-Tsuchiya Reiko , Tokuda Emi , Saji Shigehira 
Published: June 2023

2558 Background: Given the function of human leukocyte antigen (HLA) in immunity, variation of the HLA type could affect the development of immune-related adverse events (irAEs) that is induced by immune checkpoint inhibitors (ICIs). In this study, we examined whether HLA types of cancer patients could be a marker for the development of irAE thyroiditis. Methods: In the present case-control study, we randomly selected 20 cancer patients who received ICIs and developed thyroiditis (irAE thyroiditis group) and 51 who did not develop thyroiditis (No irAE thyroiditis group). The 51 patients were subdivided into 31 who did not develop any irAEs (No irAEs group) and 20 who developed irAEs other than thyroiditis (irAEs other than thyroiditis group). HLA types were classified at the amino acid sequence level (Field 2 resolution). Frequency of the HLA types of each group was compared. Results: There was no significant difference in HLA type prevalence, comparing the irAE thyroiditis group and the No irAE thyroiditis group. When comparing the irAE thyroiditis group with the No irAEs group, the irAE thyroiditis group had significantly higher HLA-X, HLA-Y, and HLA-Z (Tentative names are used here, and actual HLA types will be announced at the time of publication). This result suggested that irAEs other than thyroiditis in the control group could be noise. HLA-X had a specificity of 96.8% for the development of irAE thyroiditis. HLA-Y had a sensitivity of 80.0%. Patients carrying both HLA-X and HLA-Y had the highest specificity of 100.0%. There was a significant difference in frequency of HLA-Y between irAEs other than the thyroiditis group and the No irAEs group. HLA-Y may be a universal marker for any irAEs. On the other hand, HLA-X is a specific marker for thyroid irAE. The median time of onset of irAE thyroiditis in all patients was 72 days after ICI administration. Patients with both HLA-X and HLA-Y tended to have an earlier onset with a median onset time of 53 days. Conclusions: We found an association between specific HLA types and the development of irAE thyroiditis. The present study indicates that HLA types could be a predictive marker for the development of irAE thyroiditis.
The prognostic ability of immune scoring system Immunoscore in patients with localized colon cancer: A systematic review and meta-analysis.
Authors: Padayao John Lois , Dy Carlos 
Published: June 2023

2565 Background: Assessment of tumor recurrence risk as well as prognosis in resectable colon cancer are still mainly based on the histopathological criteria of tumor invasion reflected in the TNM classification. Robust data in colorectal tumor research show the value of tumor microenvironment and tumor‐infiltrating lymphocytes (TILs) as novel parameters to predict recurrence and survival. The utility of the derived tumor Immunoscore via digital pathology is now being promoted to refine the prognosis of early colon cancer patients together with the TNM staging to guide treatment decisions, but is not yet a recommendation in international consensus guidelines. This study assessed the prognostic value of this diagnostic tool in predicting clinical outcomes. Methods: A systematic literature search using PubMed, Embase, and Cochrane CENTRAL Library was conducted to identify observational or randomized controlled studies from January 2008 and December 2022 according to pre- specified inclusion and exclusion criteria which examined the relationship between Immunoscore and long-term prognosis (overall survival and disease free survival) in patients with localized colon cancer. Hazard ratios with 95% confidence interval for overall survival (OS) and disease-free survival (DFS) were extracted. Pooled estimates of the prognostic ability of Immunoscore were obtained using bi-variate random effects model. Statistical heterogeneity between studies was evaluated with I2 statistics and explored through sensitivity analysis. Results: A total of 5,718 patients from 11 studies were included in the meta-analysis. Moderate heterogeneity between studies for both DFS (I2= 27%) and OS (I2= 33%) was seen. The pooled analysis indicated that compared to those with high Immunoscore, colon cancer patients with low Immunoscore were associated with shorter disease-free survival (HR 1.75, 95% CI 1.53 - 2.49) and overall survival (HR 1.87 95% CI 1.45 – 2.13). In the subgroup analyses, this association was consistent across geographic regions and maximum time of follow up. Conclusions: A low Immunoscore was significantly associated with poorer clinical outcomes in terms of disease-free survival and overall survival. The use of Immunoscore provides valuable estimate of the risk of recurrence in patients with early colon cancer and has the potential to greatly affect treatment decisions in conjunction with the existing risk assessment system.
Uliledlimab and toripalimab combination therapy in treatment naive advanced NSCLC: Phase 1b/2 clinical trial results using CD73 as a potential predictive biomarker.
Authors: Zhou Qing , Wu Lin , Jiang Bo , Du Yingying , Wang Buhai , Hu Xiaohua , He Yong , Zhao Mingfang , Yao Yu , Cui Jiuwei , Dong Xiaorong , Zhang Haibo , Ma Rui , Wang Huijuan , Zhang Jian , Xia Jinjing , Zhang Yanni , Hayslip John W. , Zhu Andrew X. , Wu Yi-Long 
Published: June 2023

2570 Background: CD73 is a rate-limiting enzyme in adenosine pathway promoting immune suppression in tumor microenvironment. The role of CD73 as a novel target in cancer immune therapy is further supported by correlation between its high expression in tumors and poor prognosis in cancer pts. Here we evaluate safety and efficacy of uliledlimab, a differentiated CD73 antibody, in combination with toripalimab, a PD-1 antibody, in treatment naive NSCLC pts. In particular, we investigate the correlation between clinical response and tumor CD73 expression to assess its value as a potential predictive biomarker, following a previous study where a correlation was indicated (Robert F, et al. ASCO 2021). Methods: Pts with treatment naïve advanced NSCLC (EGFR/ALK wild type) were enrolled in a dose-expansion cohort and received uliledlimab (20mg/kg or 30mg/kg Q3W) in combination with toripalimab 240mg Q3W. Safety, the primary endpoint, and clinical efficacy including ORR, determined by iRECIST, were evaluated. Baseline CD73 and PD-L1 expression in tumor specimens were measured by IHC. The cut-off value for CD73High expression was assessed by ROC analysis for correlation with ORR. Results: As of 12/2/22, 66 pts were enrolled (median age 62.5 years, ECOG PS 0 40.6%, male 84.8% and never smokers 22.7%) and received at least one dose. Treatment-related adverse events (TRAEs) occurred in 86.4% of pts (Gr≥3, 15.2%) with no grade 5 TRAE. One patient was discontinued due to TRAE (1.5%). Efficacy was analyzed in 48 pts who were enrolled at least 90 days prior to data cut-off, allowing for two assessments. Baseline PD-L1 and CD73 expression was available in 45 pts. 40% of tumor or immune cells with ≥1+ staining intensity by IHC was determined as a preliminary cut-off for CD73High (the area under ROC curve =0.72, p = 0.02). While ORR and DCR were 31.3% (15/48) and 79.2% (38/48) respectively in all pts, ORR appeared higher in CD73High pts (50%, 9/18) and much lower in CD73Low pts (14.8%, 4/27). Highest ORR was found in a cohort with CD73High and PD-L1 TPS≥1% (57.1%, 8/14). Conclusions: Uliledlimab and toripalimab combination therapy was well-tolerated in treatment naïve advanced NSCLC pts. Clinical response of the therapy seems to correlate with high tumor CD73 expression, indicating a potential value of CD73 as a predictive biomarker. The study warrants a biomarker-guided randomized clinical trial that is currently in preparation. Clinical trial information: NCT04322006 .
Enhancing STING activation by cyclic dinucleotide-manganese particles for systemic cancer immunotherapy.
Authors: Zhou Xingwu , Sun Xiaoqi , Gong Wang , Lei Yu Leo , Moon James 
Published: June 2023

2571 Background: Activating the innate immune pathway of stimulator of interferon genes (STING) can elicit potent anti-tumor immunity via the production of type-I interferons (IFN-I). However, cyclic-dinucleotide (CDN) STING agonists have undesirable pharmacological properties. Most CDNs in clinical development are administered intratumorally, thus precluding their applications in treating metastasis, hematologic malignancies, and hard-to-reach solid tumors. Here, we report the development of a new nanoparticle system for the systemic delivery of CDN STING agonists with potent efficacy, favorable pharmaceutical properties, and acceptable safety profiles. Methods: Nutritional metal ions play crucial roles in modulating immune pathways, which could potentially be used for enhancing STING activation. We screened different metal ions and found that manganese ion (Mn2+) achieved strong synergy with CDN STING agonists for inducing IFN-I production. We further developed a new nanoparticle system for co-delivery of Mn2+ and CDN for systemic cancer immunotherapy. Results: Mn2+ induced up to 77-fold enhancement in CDN-mediated STING activation. Mixing CDNs with Mn2+ resulted in the self-assembly of nanocrystals, and the lipid layer coating led to uniform nanoparticles. The resulting STING-activating CDN-Mn nanoparticle (SNP) given intravenously elicited IFN-I responses, attenuated the suppressive functions of myeloid cells, remodeled the immunosuppressive tumor microenvironment, and expanded CD8+ T cells for sustained tumor control. Intravenous SNP therapy resulted in robust anti-tumor efficacy in multiple murine tumor models, including a genetically engineered mouse model of MMTV-PyMT triple-negative breast cancer. We have also demonstrated the robust anti-tumor efficacy of SNP against VX2 head and neck squamous cell carcinomas (HNSCC) in New Zealand white rabbits. Mechanistically, the efficacy of intravenous SNP therapy was dependent on the host STING expression but independent of the tumor STING expression. In addition, the anti-tumor efficacy of SNP was decreased in mice lacking Ifnar1 or Ifngr1 expression in the B16F10 melanoma model, showing the crucial effects of IFN-I and IFN-II responses. Lastly, SNP was effectively uptaken by human PBMCs and elicited IFN-I responses when injected into fresh human tumor biopsies derived from HNSCC patients. Conclusions: SNP empowers highly effective systemic cancer immunotherapy via nanotechnology. It also underscores the potential of utilizing metal ions for immune-mediated disease treatment.