bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 26: An HLA-class II restricted HPV18 E7 specific TCR cloned from a long-term surviving cervical cancer patient
Authors: Long Jianting , Chen Xihe , Ou Shudan , Ma Minjun , Chu Junjun , Han Yanyan 
Published: March 2024

Abstract
T cell receptor (TCR)-engineered T cell therapy is a promising approach for the treatment of solid malignancy, with multiple clinical trials reporting impressive responses using affinity-matured TCRs. However, artificially mutated TCRs can increase the risk of off-target toxicities due to unexpected cross-reactions, which limits the overall safety of this approach. To tackle this challenge, we have developed a novel strategy for cloning tumor-specific endogenous therapeutic TCRs from long-term surviving patients who have responded to immunotherapy. Specifically, we have identified a TCR (10F04) that is specific to human papillomavirus type 18 (HPV18) E784-98 and restricted to human leukocyte antigen (HLA) class II molecular DRA/DRB1*09:01 from the peripheral blood of a patient with HPV-positive metastatic cervical cancer who received multiple antigens stimulating cellular therapy (MASCT) and exhibited sustained T-cell immunological responses. Upon transduction into human T cells, the 10F04 TCR demonstrated robust antitumor activity in both in vitro and in vivo models. Notably, the TCR effectively redirected both CD4 and CD8 T cells to specifically recognize tumor cells and induced multiple cytokine secretion along with durable antitumor activity. Importantly, no cross-reactivity was detected in toxicity studies. As a result, this TCR is currently being investigated in a phase I clinical trial for treating HPV18-positive cancers (NCT05787535). The first-in-human infusion had been just completed. No severe adverse event had been observed, while an early proliferation of TCR-T was detected several days after infusion. Our approach of cloning tumor-specific and safe endogenous therapeutic TCRs from long-term surviving patients offers a promising strategy for developing safe and effective TCR-engineered T-cell therapies for solid tumors. These findings provide important insights into the potential of HLA class II-restricted TCR-T therapy as a novel cancer treatment modality.
Citation Format: Jianting Long, Xihe Chen, Shudan Ou, Minjun Ma, Junjun Chu, Yanyan Han. An HLA-class II restricted HPV18 E7 specific TCR cloned from a long-term surviving cervical cancer patient [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 26.
Abstract 2606: Targeting mesothelin on tumor cells with a novel, completely human fusion construct delivering the serine protease granzyme B
Authors: Mohamedali Khalid A. , Cheung Lawrence H. , Iurascu Mihai G. , DePalatis Louis R. , Thuning-Roberson Claire , Rosenblum Michael G. 
Published: March 2024

Abstract
Mesothelin is a cell-surface antigen identified as an excellent cancer-associated target upregulated on numerous tumors, including pancreatic and ovarian cancers, as well as mesotheliomas and lung cancers. Granzyme B is a highly cytotoxic serine protease nominally delivered by immune effector cells to target cells, resulting in induction of cell death pathways that are both caspase-dependent and -independent. We developed a fusion protein (GrB-Fc-SD1) composed of GrB and containing SD1, a novel human single-domain antibody that uniquely targets Region III of the mesothelin glycoprotein. The SD1 and GrB domains are tethered by a human IgG heavy chain fragment which enables dimerization. The fusion protein was transiently expressed in HEK-293E cells in serum-free conditions with a final yield of 5 mg/L. Western blot analysis confirmed the expression of this construct as a homodimer. The GrB enzymatic activity in GrB-Fc-SD1 was similar to that of commercially available human granzyme B. ELISA studies confirmed the specific recognition of the SD1 antibody to mesothelin extra-cellular domain. Biacore analysis confirmed binding of GrB-Fc-SD1 with a dissociation constant between 10−7 and 10−8 M. In vitro cytotoxicity against a limited panel of lung, pancreatic and ovarian cancer cell lines expressing surface mesothelin indicate cytotoxicity in the nanomolar range, while cytotoxicity against mesothelin-negative cells was at micromolar levels. Ex vivo serum stability studies showed excellent stability of GrB-Fc-SD1 at both 4°C and -20°C over ninety days and a half-life of approximately three days at elevated temperatures (37°C). In vivo efficacy studies against Capan-2 xenografts in nude mice using both weekly x 5 and QOD x 5 doses of GrB-Fc-SD1 (130 mg/mg total dose, delivered IP) indicated highly significant efficacy in both dose regimens, with p &lt; 0.001 compared to vehicle-treated mice. Mouse body weights were not impacted at the efficacy doses used in this study, indicating that this dose was non-toxic. Histopathology studies revealed a significant decrease in the percentage of Ki-67+ nuclei in treated tumors compared to controls. These data warrant continued investigation of the protein as a therapeutic candidate. Research supported by H2Biologics. Research conducted, in part, by the Clayton Foundation for Research.
Citation Format: Khalid A. Mohamedali, Lawrence H. Cheung, Mihai G. Iurascu, Louis R. DePalatis, Claire Thuning-Roberson, Michael G. Rosenblum. Targeting mesothelin on tumor cells with a novel, completely human fusion construct delivering the serine protease granzyme B [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 2606.
Abstract 2609: Vesicular LGALS3BP is a neuroblastoma biomarker and a therapeutic target for combination therapy with antibody-drug conjugate and checkpoint inhibitors
Authors: Sala Gianluca , Capone Emily , Cela Ilaria , Lovato Giulio , Lamolinara Alessia , Iezzi Manuela , Rossi Cosmo , De Laurenzi Vincenzo , Ippoliti Rodolfo , Iacobelli Stefano 
Published: March 2024

Abstract
We have recently shown that targeting circulating LGALS3BP with a maytansinoid-based non-internalizing antibody-drug conjugate (ADC) resulted in tumor eradication of high-risk NMYC-amplified neuroblastoma patient-derived xenografts. LGALS3BP is a high molecular weight, hyperglycosylated and secreted protein which is enriched at the surface of cancer-derived extracellular vesicles (EVs). Here, using a neuroblastoma pseudometastatic preclinical model, we extended our previous work demonstrating that EVs-associated LGALS3BP strictly correlates with disease progression and dissemination. Moreover, to investigate whether LGALS3BP ADC, named 1959sss/DM4, possesses immune-modulating properties, we analyzed DM4 ability to induce immunogenic cell death (ICD). Interestingly, using NXS2 murine neuroblastoma cells, we found that DM4 induced a time-dependent surface expression of three different ICD marker, i.e. calreticulin, HSP70 and HSP90. As it was recently demonstrated that to limit immune surveillance NXS2 cells increase PD-L1 expression in response to IFNγ production in the tumor immune infiltrate, we evaluated potential synergism between LGALS3BP targeting ADC and an anti-PD1 immune check-point inhibitor. To this aim, we genetically manipulated NXS2 cells which resulted nearly negative for murine LGALS3BP expression. Therefore, using a lentivirus-based gene transduction system, we obtained NXS2 cells producing large amounts of vesicular human LGALS3BP. Of note, the vesicular LGALS3BP could be quantified by ELISA and imaged by confocal microscopy using 1959 antibody. Strikingly, while single agent treatment showed a limited antitumor activity in NXS2-LGALS3BP syngeneic model, the LGALS3BP ADC/anti-PD1 combination resulted in a marked inhibition of tumor growth, which was followed by a prolonged survival. In conclusion, our findings establish that vesicular LGALS3BP is a potential biomarker for liquid biopsy and reveals this protein as a suitable target for therapeutic strategy that combines 1959sss/DM4 ADC with immune checkpoint blockade for the treatment of LGALS3BP expressing neuroblastoma.
Citation Format: Gianluca Sala, Emily Capone, Ilaria Cela, Giulio Lovato, Alessia Lamolinara, Manuela Iezzi, Cosmo Rossi, Vincenzo De Laurenzi, Rodolfo Ippoliti, Stefano Iacobelli. Vesicular LGALS3BP is a neuroblastoma biomarker and a therapeutic target for combination therapy with antibody-drug conjugate and checkpoint 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 2609.
Abstract 2637: Super-enhancer-related m6A modification promotes immune escape in hepatocellular carcinoma associated with metabolic dysfunction-associated fatty liver disease via CCL4 signaling pathway inhibition
Authors: Xu Liangliang 
Published: March 2024

Abstract
The incidence and mortality rates of hepatocellular carcinoma caused by metabolic dysfunction-associated fatty liver disease (MAFLD) have been increasing in recent decades, emphasizing the urgent need for new therapeutic approaches. Emerging evidence suggests that approximately 80% of super-enhancer RNAs (seRNAs) undergo selective m6A modifications. seRNA and m6A modifications have been implicated in the progression of various cancers, making them potential targets for therapeutic intervention. Based on our previous research, we hypothesize that the lack of m6A modification on the super-enhancer of CCL4 may contribute to the reduced expression of the CCL4 gene in HCC.To validate our hypothesis, we employ nano-chromatin immunoprecipitation combined with high-throughput sequencing (nano-ChIP-seq), m6A-seq (m6A-seq), single-cell RNA sequencing (scRNA-seq) on multiple models including clinical samples, cell lines, and mouse models. Interestingly, we have made a significant discovery that overexpression of CCL4 in tumor cells enhances the infiltration of CD8+ T cells, which are crucial for anti-tumor immune responses. In contrast, normal tissue cells undergo m6A modification on the CCL4 seRNAs, while this modification is notably absent in cancerous tissue. Consequently, the reduced expression of CCL4 could impair the tumor-killing capabilities of CD8+ T cells and facilitate immune evasion by the tumor.Furthermore, our results shed light on the underlying molecular mechanisms involving super-enhancers and m6A modifications in the regulation of immune escape in MAFLD HCC. We identify the specific super-enhancer and downstream regulatory genes influenced by m6A modification in MAFLD-associated HCC, providing valuable insights and novel targets for the treatment of this condition.
Citation Format: Liangliang Xu. Super-enhancer-related m6A modification promotes immune escape in hepatocellular carcinoma associated with metabolic dysfunction-associated fatty liver disease via CCL4 signaling pathway inhibition [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 2637.
Abstract 2675: A new dual-targeting anti-immune checkpoint therapeutic approach for the treatment of “hot” and “cold” tumors
Authors: Moutuou Moutuaata M. , DeLuce Nora , Creton Elodie , Lucchi Honorine , Accolas Julien , Savatier Alexandra , Léonetti Michel 
Published: March 2024

Abstract
Anti-Immune Checkpoint (ICP) antibody approaches provided a major breakthrough in the field of cancer therapy. However, their efficiency is mainly restricted to patients with “hot” tumors characteristics, which limits the breadth of their use. To tackle this limit, we developed a new strategy aimed at amplifying anti-ICP antibody ability to activate immune cells. This approach is based on antibodies engineered to make them able to target both an ICP receptor and a co-receptor often involved in receptor-mediated activation, namely Heparan Sulfate Proteoglycan.
The dually-targeting engineered anti-ICP antibodies were produced in eukaryotic cells. They were purified to homogeneity using two chromatographic steps and remain stable for months in frozen conditions. A first engineered antibody-derived construct was assessed for its ability to impact immune cell activation and tumor-growth. It is able to stimulate subsets of immune cells from mouse splenocytes as well as from human peripheral mononuclear cells, in vitro. In addition, its injection in healthy mice triggers activation of T-cells. Last, in mice previously implanted with different types of cancer cells it impacts growth of “hot” as well as “cold” tumors. Immune profiling in a “cold” cancer model shows that the treatment decreases the proportion of MDSCs and increase that of CD8+ T-cells and NK-cells in tumor microenvironment making it close of that found in “hot” tumors. Altogether, our data indicate that our first engineered antibody-derived construct presents an attractive immunotherapeutic potential for a large variety of cancers paving the way for its evaluation in clinical phases.
Citation Format: Moutuaata M. Moutuou, Nora DeLuce, Elodie Creton, Honorine Lucchi, Julien Accolas, Alexandra Savatier, Michel Léonetti. A new dual-targeting anti-immune checkpoint therapeutic approach for the treatment of “hot” and “cold” tumors [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 2675.
Abstract 2680: Knockout on enhancer of zeste homolog 2 in triple-negative breast cancer model alters immune signaling and decreases tumor progression and lung metastasis
Authors: Monterroza Lenore , Parrilla Maria , Samaranayake Sarah , Rivera Dormarie , Dobosh Brian , Selvaraj Periasamy , Tirouvanziam Rabindra M. 
Published: March 2024

Abstract
Triple-negative breast cancer (TNBC) is an aggressive and highly metastatic subtype of breast cancer. Lack of known targets, patient-to-patient variations of target antigens and resistance to immune checkpoint inhibitors make TNBC one of the most challenging cancers to treat. TNBC is enriched in tumor-infiltrating neutrophils (TINs). TINs are key contributors to pro-tumorigenic processes linked to “immunologically cold” tumors, in part by preventing recruitment of T-lymphocytes (TILs). Prior studies identified the Enhancer of zeste homolog 2 (EZH2) as a prominent methyltransferase expressed in primary and metastatic TNBC cells. EZH2 is an upstream regulator of the stimulator of interferon genes (STING), a critical innate sensor mediating anti-tumor responses that is inhibited in cold tumors. To date, EZH2/STING signaling in TINs is relatively unknown. Here, we hypothesized that EZH2 inhibition and subsequent STING activation could exert anti-tumor activity by modulating TINs. We derived EZH2-knockout (KO) and EZH2-overexpressed (OX) cell lines from the well-established 4T1 TNBC mouse cell line. We demonstrate a 10-fold decrease in lung metastasis and significant reduction of 4T1 EZH2 KO tumor growth compared to the parent 4T1 cell line. Concomitantly, the TIN:TIL ratio was significantly reduced in 4T1 EZH2 KO vs. control tumors. Overexpression of EZH2 in 4T1 TNBC cells did not affect the primary tumor growth or lung metastasis. A therapeutic study using the repeated injection of the STING agonist MSA-2 after tumor engraftment in vivo also demonstrated a decrease in lung metastasis and tumor growth in MSA-2 vs. vehicle injection with both 4T1 parent and EZH2 KO cell lines. Together, our findings suggest a key role for STING pathway modulation and TIN/TIL poise in primary TNBC progression and lung metastasis.
Citation Format: Lenore Monterroza, Maria Parrilla, Sarah Samaranayake, Dormarie Rivera, Brian Dobosh, Periasamy Selvaraj, Rabindra M. Tirouvanziam. Knockout on enhancer of zeste homolog 2 in triple-negative breast cancer model alters immune signaling and decreases tumor progression and lung metastasis [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 2680.
Abstract 2687: NK cell plasticity in SCLC is linked to tumor transcriptional diversity
Authors: Malin Justin , Zhuang Sophie , Sharma Ajit Kumar , Clara Joseph , Thomas Anish 
Published: March 2024

Abstract
Natural Killer (NK) cells are innate lymphoid cells that are pivotal in the body’s response to cancer. To harness their cytotoxicity in clinical interventions, it is critical to understand their biology, which is highly tissue- and tumor-specific. In Small Cell Lung Cancer (SCLC), reports have described widely divergent levels of NK activity in neuroendocrine (NE) and non-neuroendocrine (non-NE) subtypes. Diminished self-antigen presentation by MHC-I is a classic trigger of NK cell activity but, counter-intuitively, NK cells exhibit reduced cytotoxicity in MHC-I downregulated NE SCLC.
To explore potential causes for depressed NK activity in NE SCLC relative to non-NE, we applied scRNA-Seq to 45 SCLC patient biopsies and rapid autopsies, and identified 4000 NK cells based on gene expression signature. There was a notable dip in NK cell infiltration in NE tumors compared to non-NE tumors. Tumor cell-surface ligands recognized by activating and, unexpectedly, inhibitory NK cell receptors were generally both more highly expressed in non-NE tumors. While inhibitory NK receptors were comparable in NE and non-NE tumors, activating NK receptors were expressed significantly, though modestly, higher in non-NE. We observed tumor site-specific gene expression patterns in NK cells from different metastatic sites. Overall, there were a remarkably high number of differentially expressed genes (&gt;1k) between NK cells from NE and non-NE tumors, even after controlling for tumor site. Together, our work suggests that NK cell plasticity is manifest at the level of SCLC transcriptional subtypes.
Citation Format: Justin Malin, Sophie Zhuang, Ajit Kumar Sharma, Joseph Clara, Anish Thomas. NK cell plasticity in SCLC is linked to tumor transcriptional diversity [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 2687.
Abstract 2756: Single cell analysis reveals treatment response-associated plastic cell state and lineage switching in aggressive variant prostate cancer
Authors: Guo Shuai , Subramani Elavarasan , Frigo Daniel E. , Pilié Patrick G. , Goswami Sangeeta , Soundararajan Rama , Aparicio Ana , Wang Wenyi 
Published: March 2024

Abstract
Prostate cancer (PCa) is a common malignancy in men over 65, ranking as the second leading cause of cancer-related deaths among men in the United States. While most prostate cancers respond to androgen receptor (AR) targeting therapies, Aggressive Variant Prostate Cancers (AVPCs), characterized by RB1, Tp53, and/or PTEN alterations, do not. AVPCs exhibit lineage plasticity in preclinical models, enabling the expression of non-prostate epithelial lineage markers to adapt to therapeutic pressures. However, a measurable definition of lineage plasticity as a potential therapeutic target for clinical trials is currently lacking. To arrive at said definition, we perform single-nuclei RNA sequencing (snRNA-seq) on lymph node metastases from twelve AVPC patients with matched sample pairs of both pre- and post-treatment. Additionally, we include snRNA-seq data from eight non-AVPC patients as a baseline. Expanding upon our prior establishment of tumor-specific total mRNA expression (TmS) as an indicator of tumor cell lineage plasticity (Nature Biotechnology, PMID: 35697807), we employ unique molecular identifier (UMI) counts from individual cells to quantify this plasticity. We reveal a distinct cluster of tumor cells with notably high UMI counts in resistant samples, indicative of a less differentiated state and increased transcriptional diversity. This high-UMI cluster is marked by significant enrichment in pathways associated with epithelial-mesenchymal transition (EMT), stemness, one-carbon metabolism, and arginine metabolism pathways. Concurrently, there is a reduction in activity within the androgen receptor (AR) and DNA damage repair pathways. These characteristics suggest an enhanced capacity for lineage switching in AVPC tumor cells, potentially contributing to evasion from targeted therapies. Moreover, immune components within resistant samples consistently display lower UMI counts, indicating a diminished propensity for lineage plasticity. In conclusion, we present a better understanding of lineage switching in therapeutic resistance in AVPCs, paving the way for novel treatment strategies in PCa management.
Citation Format: Shuai Guo, Elavarasan Subramani, Daniel E. Frigo, Patrick G. Pilié, Sangeeta Goswami, Rama Soundararajan, Ana Aparicio, Wenyi Wang. Single cell analysis reveals treatment response-associated plastic cell state and lineage switching in aggressive variant prostate cancer [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 2756.