bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 492: Investigating solute carrier SLC46A3 to enhance nanotherapeutic delivery for cancer
Authors: Kenny-Serrano Julianna , Nelson Elizabeth B. , Covarrubias Gil , Nabar Namita , Straehla Joelle , Gupta Aanchal , Boehnke Natalie , Hammond Paula
Published: March 2024

BACKGROUND: Nanoparticle therapeutics have multifaceted potential to improve efficacy and decrease toxicity of cancer treatments. Nanotherapeutics can encapsulate a range of cargos and have tunable surface chemistry that can enhance drug delivery to tumors. Cellular features also play a role in successful drug delivery, and predictive biomarkers may improve patient selection for clinical trials. In previous work, we identified SLC46A3, a poorly characterized lysosomal solute carrier, as a strong negative regulator of lipid-based nanoparticle uptake across many cancer models. We hypothesize that the expression of SLC46A3 will act as a predictive biomarker for therapeutic nanoparticles.

METHODS: To test this hypothesis, we used two engineered model systems to probe the relationship between SLC46A3 expression and therapeutic response: lentiviral overexpression in a natively low-expressing melanoma model and genetic deactivation in a natively high-expressing breast cancer model. A library of FDA-approved and experimental therapeutic nanoparticles were screened in both models under a range of conditions. Liposomal formulations of chemotherapeutics were included with high and low surface functionalization of polyethylene glycol to investigate the impact of this ubiquitous ‘stealth’ polymer on our biomarker of interest. Solid lipid nanoparticles with nucleic acid cargo were also included. Orthogonal methods were used to probe these interactions and the therapeutic efficacy of nanotherapeutics, including pulsed dosing viability assays and label-free live-cell imaging.

RESULTS: We found that SLC46A3 expression is a class-specific biomarker of lipid-based nanoparticle uptake and is predictive of distinctive patterns of response to therapy-loaded nanoparticles in both formulation and time-dependent manners. Additionally, we present a new technology for long-term, live cell imaging of nanoparticle uptake and trafficking that can discriminate mechanisms of cytotoxicity, accelerating the study of nano-bio interactions. These findings highlight a promising approach for the functional validation of nanoparticle-specific biomarkers using clinically relevant formulations that have the potential for rapid repurposing.

Citation Format: Julianna Kenny-Serrano, Elizabeth B. Nelson, Gil Covarrubias, Namita Nabar, Joelle Straehla, Aanchal Gupta, Natalie Boehnke, Paula Hammond. Investigating solute carrier SLC46A3 to enhance nanotherapeutic delivery for 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 492.
Abstract 494: Fluorescent carbon dots: A novel bioimaging tool to reveal the mechanism of action of anticancer drugs in cells
Authors: Clermont-Paquette Adryanne
Published: March 2024

The mechanism of action of many cancer drugs at the subcellular level remains unknown. Yet, this knowledge could be critical in driving structure-activity-relationship studies to improve their efficacy and selectivity. To gain a deeper understanding of the mechanisms governing the action of anticancer drugs within cells, we are developing an innovative methodology to visualize drug interactions. Carbon dots (CDs) are carbon-based nanoparticles, distinguished not only by their remarkable optical properties but also by their minimal cytotoxicity, rendering them highly suitable for bioimaging applications. We recently found that amine-passivated CDs with a negative surface charge and amphiphilic properties were enriched in the cytosol of cultured human cells. These CDs emit fluorescence in the blue spectra with high quantum yields and are not cytotoxic. Importantly, their surface is decorated with amine functional groups that can be used for coupling to polyethylene glycol to create a linker that can be covalently attached to drugs of interest. First, we are covalently linking amine-passivated CDs to Doxorubicin (Dox), a well-known anti-cancer drug that intercalates DNA and also has fluorescence properties that differ from the CDs. This will permit us to monitor and validate the conjugation of Dox to CDs, their cellular uptake and localization. The next step will be to use this approach to reveal the cellular mechanism of action of novel anticancer drugs. Specifically, our group has rationally designed thienoisoquinoline compounds with selectivity and high efficacy for triple negative breast cancer cells. In vitro they disrupt microtubule polymerization in the nanomolar range. However, in cells they cause mitotic arrest and spindle phenotypes that are distinct from colchicine, another microtubule-targeting drug. Our approach will reveal if these compounds target the centrosomes and/or microtubules and if this varies with cell types. This innovative approach could yield profound insights into the intracellular mechanisms of anticancer drugs to facilitate structure-activity-relationship studies, to improve their efficacy and selectivity for drug development.

Citation Format: Adryanne Clermont-Paquette. Fluorescent carbon dots: A novel bioimaging tool to reveal the mechanism of action of anticancer drugs in cells [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 494.
Abstract 5038: Validation of a comprehensive next-generation sequencing liquid biopsy assay for clinical diagnostics and clinical trial applications
Authors: Tan Xin-Xing , Peters Tricia , Berry Meredith , Barber Kimberly , Jeter Kristyn , Lovell Brigitte , Elizondo Paula , Mou Yanglong , Thomas Brad , Xie Heng , Guo Jiannan
Published: March 2024

Liquid biopsy next generation sequencing (NGS) provides a powerful non-invasive approach to detect solid tumor derived somatic variants in a massively parallel manner. Liquid biopsy NGS has rapidly become an invaluable choice of testing technologies for clinical utility where invasive tissue biopsy is inaccessible and/or inadequate for molecular characterization. We present here a targeted NGS gene panel assay designed and validated specifically for liquid biopsy characterization. The assay was designed to test cell free DNA and cell free RNA from blood plasma for detection of single nucleotide variants (SNV), insertion/deletion (Indel), copy number variants (CNV), and gene fusions present in 52 most commonly mutated genes in cancers. The validation was performed to assess assay accuracy, specificity, sensitivity, repeatability, and reproducibility, with including pre-characterized reference controls, retrospective plasma samples from cancer patients, and prospectively-collected clinic blood specimen. Triplicates were included and testing was performed at different times and by different operators to assess the assay precision. Over 100 clinic blood specimens were collected in Streck cfDNA tubes from multiple trial centers, including treatment naïve phase III-IV patients with solid tumor (lung cancer, breast cancer, brain cancer, prostate cancer, kidney cancer, melanoma, etc) as well as heathy donors, processed within 7 days for plasma separation and total nucleotide acid (TNA) extraction, and assessed by the NGS assay for mutation detection. The identified mutations and wild-types were verified with orthogonal ddPCR. Assay sensitivity as low as 0.1% for SNV and Indel, 1.3 fold change for CNV, and 10 copies for gene fusion were observed, with near-perfect assay specificity (>99.99%) at these sensitivity levels. In conclusion, a targeted NGS liquid biopsy assay was analytically validated under medical oversight, with a demonstrated rigor of the test by its high sensitivity/specificity and robust reproducibility. The validated assay has been utilized for clinical diagnostics and to support research and development efforts in clinical trials.

Citation Format: Xin-Xing Tan, Tricia Peters, Meredith Berry, Kimberly Barber, Kristyn Jeter, Brigitte Lovell, Paula Elizondo, Yanglong Mou, Brad Thomas, Heng Xie, Jiannan Guo. Validation of a comprehensive next-generation sequencing liquid biopsy assay for clinical diagnostics and clinical trial applications [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 5038.
Abstract 5119: Exploring the relationship between metabolism and the epigenome in MYCN-amplified neuroblastoma
Authors: Dalton Krista M. , Lochmann Timothy L. , Coon Colin , Calbert Marissa L. , Dozmorov Mikhail , Shock Lisa , Koblinski Jennifer , Faber Anthony C.
Published: March 2024

Background: Neuroblastoma (NB) is the most common cancer of infancy and is responsible for the most cancer-related deaths in children under the age of five. The transcription factor MYCN is the most studied oncogene in NB and automatically confers high-risk. MYCN drives numerous effects in the cell, including metabolic changes that are critical for oncogenesis. NB has also been demonstrated to be a highly methylated cancer, with DNA methyltransferases (DNMTs) largely responsible for these modifications. Epigenetics and metabolism are closely connected, with oncogene-driven metabolic rewiring modifying the epigenetic landscape through increases of metabolites that are critical for the activities of epigenetic enzymes, and conversely the enhanced epigenetic activity further regulating the activity of metabolic enzymes.

Aims: As metabolism is believed to be a key regulator of the epigenome, the objective of this proposal is to understand the connection between MYCN-DNMT1-driven epigenetic alterations and dysregulated metabolism in MYCN-amplified NB.

Methods: RNA-seq and mass spectrometry analyses were performed to explore the MYCN-driven metabolome in NB and changes that occur with DNMT1 inhibition. The efficacy of DNMT1 inhibition in MYCN-amplified NB was tested in vivo.

Results: Our data demonstrate that sensitivity to DNMT1 inhibition is linked to MYCN status in NB. We found that MYCN indeed dysregulates metabolism in NB, and DNMT1 inhibition led to a global decrease in oxidative phosphorylation and exacerbated mitochondrial dysfunction in MYCN-amplified NB. Expression of MYCN was linked to an increase in methionine and subsequently SAM, the universal methyl donor, further supporting reliance on DNMT1. Most importantly, we have found that treatment with DNMT1 inhibitors in vivo inhibited tumor growth, even after treatment had stopped.

Conclusion: We have found a correlation between MYCN and metabolism that supports DNMT1 activity. This relationship allows for a potential treatment for a high-risk disease in which there is few clinically actionable targets.

Citation Format: Krista M. Dalton, Timothy L. Lochmann, Colin Coon, Marissa L. Calbert, Mikhail Dozmorov, Lisa Shock, Jennifer Koblinski, Anthony C. Faber. Exploring the relationship between metabolism and the epigenome in MYCN-amplified neuroblastoma [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 5119.
Abstract 5147: Identification of pharmacodynamic biomarkers in PBMCs and cancer cells for SIK2 kinase inhibitor therapy
Authors: Yang Hailing , Mao Weiqun , Rask Philip , Peter Markus , Swaminath Gayathri , Morris Stephan W. , Lu Zhen , Bast Robert C.
Published: March 2024

Salt-inducible kinase 2 (SIK2) is an AMPK family kinase involved in multiple signaling pathways including the LBK1-SIK-HDAC, LKB1-SIK-CRTC/CREB, PI3K-PKA-mTOR and Hippo pathways. The expression of SIK2 is significantly upregulated in fraction of ovarian and breast cancers, and associated with poor prognosis. Inhibition of SIK2 kinase activity suppresses growth of ovarian and breast cancers as well as progression of acute myeloid leukemia. SIK2 inhibitors synergistically enhance sensitivity of ovarian and breast cancer cells to paclitaxel, carboplatin and PARP inhibitors. Utilization of SIK2 inhibitors to treat cancers has been investigated in preclinical studies and in a Phase I clinical trial (NCT04711161). To enable pharmacodynamic (PD) analysis in clinical studies, we have developed biomarkers that measure the effect of GRN-300, a SIK2/3 inhibitor, on SIK2 signaling in ovarian cancer cells. Class IIa histone deacetylases (HDAC4/5/7) are SIK2 substrates in cancer cells; and are also expressed in peripheral blood mononuclear cells (PBMCs). We found that GRN-300 decreased phosphorylated HDAC4/5/7 levels in ovarian and breast cancer cells (Lu et al. J Clin Invest, 2022) and now report that this SIK2 inhibitor also decreases pHDAC4/5/7 in PBMCs ex vivo in a dose-dependent manner using Western blot analysis. We also identified three mRNA markers in PBMCs by RNAseq analysis - SEPP1, RNASE1 and MS4A6A - which are up-regulated by GRN-300 in PBMCs isolated from healthy donors. However, these mRNA markers were either expressed at very low levels or not dramatically changed in cancer cells. We identified an additional mRNA marker panel using RNAseq technology by screening seven ovarian and six breast cancer cell lines: A2780, Caov3, IGROV1, OC316, OVCAR3, OVCAR8, SKOv3ip, BT549, Cal51, HCC1937, MDA-MB-231, MDA-MB-436 and MDA-MB-468. mRNA expression of three genes - DDIT4, NUPR1 and IL6 - is upregulated by GRN-300 in more than 9 of 13 cell lines tested. Five genes - ID1, ID2, ID3, ID4 and TGFBI - are down-regulated in response to GRN-300 treatment in more than 10 of 13 cell lines. The decrease in HDAC4/5/7 phosphorylation, and alterations in mRNA markers we discovered could serve as PD biomarkers for clinical trials to evaluate the effects of SIK2 inhibitors and provide insights in the mechanisms of drug action.

Citation Format: Hailing Yang, Weiqun Mao, Philip Rask, Markus Peter, Gayathri Swaminath, Stephan W. Morris, Zhen Lu, Robert C. Bast. Identification of pharmacodynamic biomarkers in PBMCs and cancer cells for SIK2 kinase inhibitor therapy [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 5147.
Abstract 5162: Tumor transcriptional identity as a key predictor of clinical outcome in EGFR-Mutant non-small cell lung cancer
Authors: Heeke Simon , Nilsson Monique , Gibbons Don , Zhang Jianjun , Wistuba Ignatio I. , Park Keunchil , Heymach John V. , Le Xiuning
Published: March 2024

Background: EGFR tyrosine-kinase inhibitors (TKIs) produce significant clinical benefit in patients (pts) harboring EGFR-activating mutations, however, patient outcomes remain heterogeneous, highlighting the need to identify novel biomarkers for treatment response prediction. We aim to investigate the association of tumor transcriptional identity with clinical outcomes in EGFR-mutant NSCLC.

Methods: We used consensus classification to derive unsupervised transcriptionally distinct clusters in EGFR-mutant NSCLC from the TCGA (N= 44). Then, we built a predictive model using extreme-gradient boosting to (1) classify an additional 88 surgically resected tumors from the LCCS study and analyzed for overall survival (OS) prediction; (2) classify metastatic EGFR-mutant NSCLC from the RELAY trial with available RNAseq data (N = 106) and analyzed for progression-free survival (PFS) with erlotinib treatment.

Results: Consensus clustering revealed two distinct clusters, one associated with more lineage-preserving features and one with more lineage-independent features that could transcriptionally distinguish EGFR-mutant lung cancer. No differences in primary EGFR mutations were detected across the two clusters. Ingenuity Pathway Analysis revealed TGFB1 as the most significantly enriched pathway in the lineage-independent cluster which was also significantly more mesenchymal (p = 0.0028). For surgically resectioned EGFR-mutant NSCLC, OS was consistently shorter in the lineage-independent cluster for TCGA (23.6 vs 58.3 mo; p = 0.049; HR = 0.4) and the LCCS cohort (55.7 vs not-reached; p = 0.0073; HR = 0.27). In the metastatic setting with erlotinib treatment, there was a trend for PFS to be shorter in the lineage-independent cluster (15.2 vs 19.4 mo; p = 0.12; HR = 0.61).

Conclusions: Transcriptional identity in EGFR-mutant lung cancer are prognostic of clinical outcome. The lineage-independent tumors have inferior benefit to EGFR TKI as well as inferior OS, indicating that transcriptional biomarkers can be independent of oncogene biomarkers. These transcriptional biomarkers may help identify pts with poor outcome for future intensification of therapies.

Citation Format: Simon Heeke, Monique Nilsson, Don Gibbons, Jianjun Zhang, Ignatio I. Wistuba, Keunchil Park, John V. Heymach, Xiuning Le. Tumor transcriptional identity as a key predictor of clinical outcome in EGFR-Mutant non-small cell lung 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 5162.
Abstract 5208: Dynamic changes in circulating tumor DNA and T cell receptor repertoire predict disease progression in patients with unresectable esophageal squamous cell carcinoma
Authors: Zhang Baidong , Zhan Xiang , Li Qiang , Wang Chenxuan , Pang Jiaohui , Zhang Yaru , Wang Song , Ou Qiuxiang , Tang Haimeng , Feng Alei , Yang Zhe
Published: March 2024

Background: Patients with unresectable esophageal squamous cell carcinoma (ESCC) showed variations in their response to treatments and experienced inevitable disease progression. The development of non-invasive and clinically viable biomarkers is urgently needed. Therefore, this study aimed to investigate prognostic value of circulating tumor DNA (ctDNA) and T cell receptor (TCR) repertoire in these patients.

Methods: A total of 21 ESCC patients who underwent definitive chemoradiotherapy, radiotherapy, or immune checkpoint inhibitor (ICI)-combined therapy from October 2020 to March 2022 were retrospectively evaluated. Baseline tumor samples were collected for genomic profiling using targeted NGS. Baseline and post-therapy plasma were collected for genomic and TCR profiling.

Results: We found that ctDNA detection rates were significantly reduced after treatment (P<0.001). Post-treatment ctDNA positivity were associated with worse local recurrence-free survival (P=0.001) and progression-free survival (P=0.001), but not overall survival. Moreover, TCR clonality was significantly increased after treatment, including CDR3 diversity (P=0.003), TCR clonotype (P=0.004), and VJ pairs (P=0.010), indicating an increased T cell expansion and complexity. Specifically, we identified two increased and four reduced VJ rearrangements after treatment, and the four reduced VJ rearrangements (Vβ12-2Jβ2-4, Vβ6-9Jβ1-5, Vβ6-7Jβ1-5, and Vβ6-9Jβ1-1) were significantly associated with worse progression-free survival.

Conclusion: Our findings suggest the prognostic implication of plasma ctDNA and TCR profiles in patients with unresectable ESCC receiving definitive therapy, supporting the future development of non-invasive monitoring of disease progression in these patients.

Citation Format: Baidong Zhang, Xiang Zhan, Qiang Li, Chenxuan Wang, Jiaohui Pang, Yaru Zhang, Song Wang, Qiuxiang Ou, Haimeng Tang, Alei Feng, Zhe Yang. Dynamic changes in circulating tumor DNA and T cell receptor repertoire predict disease progression in patients with unresectable esophageal squamous cell carcinoma [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 5208.
Abstract 5217: Genome instability in renal tumors
Authors: Cho Hanbyul , Li Ginny X. , Mannan Rahul , Hosseini Noshad , Hsiao Yi , Zhang Yuping , Dagar Aniket , Kumar-Sinha Chandan , Cieslik Marcin P. , Dhanasekaran Saravana M. , Nesvizhskii Alexey I. , Chinnaiyan Arul M. , Consortium Clinical Proteomic Tumor Analysis
Published: March 2024

Genome instability (GI), a recognized cancer hallmark, encompasses CNV burden, tumor mutational burden (TMB), and microsatellite instability and is variably associated with several clinical/prognostic features in solid tumors. GI and intratumor mutational heterogeneity in clear cell renal cell carcinoma (ccRCC) were associated with poor prognosis in a large multi-site sequencing study. As 10-15% of renal tumors exhibit copy number burden associated GI, the development of proteogenomic biomarkers to identify this subset of tumors will be of high clinical value. In our in-depth analysis of CPTAC renal cancer cohorts namely, ccRCC discovery and rare RCC, we first estimated absolute copy number using CNVEX TPO analysis pipeline. Copy number burden-based genome instability was estimated by calculating whole genome instability index (wGII) from DNA sequencing data, in addition to the estimation of genome ploidy and purity of the tumors. Genome instability was associated with survival outcomes in both ccRCC and rare RCC (Ref. 1 and 2). Differential proteogenomic analysis identified IGF2BP3, IGF2BP1, PYCR1 RNA and protein, to be upregulated among high wGII tumors. High wGII tumors showed upregulation of cell proliferation and other cell cycle/proliferation concepts in gene set enrichment analysis. Interestingly, transcriptional module analysis revealed transcription factors and target genes specifically upregulated in GI cases and this pattern showed certain degree of overlap to 9p loss ccRCC tumors (another molecular ccRCC subset that is also associated with poor prognosis). Differential expression analysis also identified, kinases, phosphosites and co-regulated kinase-substrates that are dysregulated in genome instable renal tumors. IGF2BP3 immunohistochemistry staining showed specific overexpression in high wGII tumors and the high positivity was observed exclusivity in the tumor cells. Prognostic biomarkers IGF2BP3, IGF2BP1, PYCR1, and UCHL1 show great promise to be developed as clinical prognostic assays and warrants additional characterization.

Citation Format: Hanbyul Cho, Ginny X. Li, Rahul Mannan, Noshad Hosseini, Yi Hsiao, Yuping Zhang, Aniket Dagar, Chandan Kumar-Sinha, Marcin P. Cieslik, Saravana M. Dhanasekaran, Alexey I. Nesvizhskii, Arul M. Chinnaiyan, Clinical Proteomic Tumor Analysis Consortium. Genome instability in renal 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 5217.
Abstract 5298: BSI-093, a best-in-class anti-BTN3A monoclonal antibody for cancer immunotherapy by activation of Vg9Vd2 T cells
Authors: Dai Wenwen , Li Jun , Liu Jinyu , Li Hongyan , Liu Xiaodong F. , Xia Shukai , Lyu Qun , Davis Hugh M. , Chen Mingjiu , Peng Zeyu
Published: March 2024

Background: Gamma delta T (γδ T) cells are potent anti-cancer effectors with the potential to target tumors broadly, independent of patient-specific neoantigens or human leukocyte antigen background. Among tumor-infiltrating lymphocytes, the presence of γδ T cells is the most favorable prognostic marker for overall survival of patients with various solid and hematologic tumors. Vγ9Vδ2 T cells are the predominant γδ T cells in peripheral circulation. Butyrophilin 3A (BTN3A) members, widely expressed in tumors, have emerged as novel molecules modulating the function of Vγ9Vδ2 T cells. Targeting BTN3A for activation of Vγ9Vδ2 T cells provides an alternative strategy for cancer immunotherapy.

Experimental procedures: BALB/c mice were immunized with recombinant BTN3A1-ECD-Fc. The Biosion proprietary H3 (High-throughput, High-content and High-efficiency) antibody discovery platform was used to identify an anti-BTN3A monoclonal antibody lead candidate - BSI-093. The target binding specificity, binding activity, and affinity of BSI-093 were evaluated by protein-based ELISA, cell-based FACS and Biacore-based SPR. Ex vivo assays were used to evaluate the cellular bioactivity of BSI-093 on activating Vγ9Vδ2 T cells for killing activity of various tumor cells. B-NDG mice bearing Jurkat-GFP-Luc tumors were used to evaluate the tumor inhibitory activity of the anti-BTN3A antibody.

Summary: BSI-093 is a humanized monoclonal antibody with the following critical properties: (1) binds to all BTN3A members (BTN3A1, BTN3A2 and BTN3A3) with high affinity; (2) has potentially longer half-life and silenced Fc-effector function by Fc-engineering; (3) exhibits cross-reactivity to rhesus BTN3A members; (4) demonstrates strong activity on activating Vγ9Vδ2 T cells to kill a variety of natural cancer cells in ex vivo assays; and (5) shows significant anti-tumor efficacy in a CDX model.

Conclusion: BSI-093 demonstrates potential best-in-class biophysical properties and functional characteristics, supporting the initiation of development activities including manufacturing and IND-enabling studies.

Citation Format: Wenwen Dai, Jun Li, Jinyu Liu, Hongyan Li, Xiaodong F. Liu, Shukai Xia, Qun Lyu, Hugh M. Davis, Mingjiu Chen, Zeyu Peng. BSI-093, a best-in-class anti-BTN3A monoclonal antibody for cancer immunotherapy by activation of Vg9Vd2 T cells [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 5298.
Abstract 5312: Agonist CD40 engineered smart immunosomes induced anti-tumor immunity, reduced associated toxicities and showed tumor free survival in murine glioblastoma model
Authors: Gaur Vidit , Tyagi Witty , Das Sanjeev , Ganguly Surajit , Bhattacharyya Jayanta
Published: March 2024

CD40 agonist antibodies (αCD40) have proven great potential as an anti-cancer immunotherapeutic by showing promising anti-tumor response in both preclinical and early clinical studies. However, it has been observed that the systemic administration of αCD40, even at low dose, is associated with serious immune- and hepato- toxicities. In addition, αCD40 showed low tumor retention and induced PD-L1 expression which makes tumor microenvironment (TME) immunosuppressive. To overcome these issues, in this study, we have developed an immunosome where αCD40 is conjugated on the surface and VJ23, a small molecule immune-modulator was encapsulated inside it . Immunosome showed higher tumor accumulation till 96 hr of administration and displayed sustained release of αCD40 in vivo. Immunosome significantly delayed tumor growth and showed tumor free survival in mice bearing GL-261 glioblastoma tumor by increasing the population of CD45+CD8+ T cells, CD45+CD20+ B cells, CD45+CD11c+ DCs and F4/80+CD86+M1 macrophage in TME. In addition, Immunosome significantly reduced the population of T-regulatory cells, M2 macrophage, and MDSCs and lowered the expression of PD-L1 to modulate immunosuppressive TME. Moreover, Immunosomes significantly enhanced the level of pro-inflammatory cytokines (IFN-γ, IL-6, IL-2) and lowered anti-inflammatory cytokines (IL-4 and IL-10) level which supported anti-tumor response. Most interestingly, Immunosomes averted the in vivo toxicities associated with free αCD40 by lowering the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), IL-6, IL-1α and reduced the degree of liver damage. Our results suggested that this novel formulation can be further explored in clinics to improve the in vivo anti-tumor efficacy of αCD40 while reducing the associated toxicities.

Citation Format: Vidit Gaur, Witty Tyagi, Sanjeev Das, Surajit Ganguly, Jayanta Bhattacharyya. Agonist CD40 engineered smart immunosomes induced anti-tumor immunity, reduced associated toxicities and showed tumor free survival in murine glioblastoma model [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 5312.
Abstract 5330: Intestine-specific homeobox edits macrophage polarization in the disease microenvironment by activating inflammasome activity
Authors: Wang Li-Ting , Hsu Shih-Hsien
Published: March 2024

Inflammasome activity has a pivotal role in the innate immune system, highlighting its importance in the development of various inflammatory diseases, including cancers. However, the specific impact of inflammasome-produced cytokines from tumor cells on disease progression remains uncertain. In this study, we show that the expression levels of inflammasome-related genes (NLRP1 and NLRP3), combined with levels of the intestine-specific homeobox (ISX), significantly correlate with factors like lesion count, disease stage, and lymph vascular invasion. Tumor cells enhance the secretion of key cytokines such as IL-1β and IL-18, exacerbating disease progression via the ISX-TWIST1 complex. In terms of mechanism, transcriptomic analysis and molecular evidence reveal that ISX interacts with the basic helix-loop-helix (bHLH) domain of TWIST1 through its homeobox domain. This interaction forms a transcriptional complex that activates inflammasomes by directly binding to the conserved sequence "-GGDWYR-" in promoter regions of several NF-κB and inflammasome-related genes (NFKBs, RELs, NLRP1, NLRP3, NEK7, AIM2, and ASC). Elevated levels of IL-18, induced by active inflammasomes, lead to the shift of M0 macrophages into the M2-like phenotype within the hepatic microenvironment. In mice with liver-specific ISX mutations, inflammasome-related genes and activities were eliminated, stopping disease progression in an acute hepatic disease model. These findings underscore the critical role of the ISX-TWIST1 complex in liver disease and tumor progression on both mechanistic and clinical levels. They also offer new insights for potential preventive and therapeutic targets in liver disease.

Citation Format: Li-Ting Wang, Shih-Hsien Hsu. Intestine-specific homeobox edits macrophage polarization in the disease microenvironment by activating inflammasome activity [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 5330.
Abstract 5340: NK-humanized mouse models for the in vivo evaluation of NK cell-based therapies in solid tumors
Authors: Rettman Pauline , Benhamouche-Trouillet Samira , Pelat Michel , Franciszkiewicz Katarzyna , Azar Georges , Blot Lydia , Calvet Loreley , Sidhu Sukhvinder , Nicolazzi Céline
Published: March 2024

Novel immunotherapeutic strategies targeting Natural Killer (NK) cells using monoclonal or multi-specific antibodies and immune modulator molecules are under development and suitable mouse models are needed to evaluate these therapeutics. We developed and characterized robust in vivo models that allow the engraftment and long-term maintenance of fully functional human NK (huNK) cells in immunodeficient transgenic mice for human cytokines (IL15). In efficacy studies, we confirmed that huNK cells are recruitable by NK cell-based therapies including therapeutic antibodies or multi-specific biologics and able to control tumor growth in these models.To establish a more relevant model for better clinical translatability for Fc-competent therapeutics evaluation, we optimized the model by validating a mouse strain with a functional knock-out of murine Fcγ receptors (FcgR-KO) to minimize the confounding impact of mouse immune cells. We characterized huNK cells kinetics and profile by flow cytometry compared to NOG-huIL15 mice. In efficacy studies with the FcgR-KO strain, therapeutic antibodies are inactive in non-NK-humanized mice, whereas in NK-humanized mice huNK cells can be recruited by the therapeutic antibody to control tumor growth by ADCC, in B cell lymphoma or multiple myeloma bearing mice.We also report the development of NK-humanized models for the evaluation of NK-based therapies in solid tumors. We characterized huNK intratumoral infiltration in several subcutaneous tumor xenograft models and evaluated the impact of tumor environment on the expression of functional NK markers (NKp46, NKG2D, Tim3, CD69) by flow cytometry and IHC studies.In summary, we have developed and optimized robust murine models sustaining fully functional huNK cells that can be proficiently recruited in vivo by various NK-based therapeutics. These models can also be used to evaluate the efficacy of combination therapies.

Citation Format: Pauline Rettman, Samira Benhamouche-Trouillet, Michel Pelat, Katarzyna Franciszkiewicz, Georges Azar, Lydia Blot, Loreley Calvet, Sukhvinder Sidhu, Céline Nicolazzi. NK-humanized mouse models for the in vivo evaluation of NK cell-based therapies in solid 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 5340.
Abstract 5342: Evaluation of autologous and allogeneic T cell-based therapies in ex-vivo and in-vivo xenograft tumor models
Authors: Liu Sherry , Borroel Jasmine , Sunico Carmen , Eastwood Emily , Corcoran Bridget , Carapia Bianca , Sperry Jantzen , Nakashima Jonathan , Shrimali Rajeev
Published: March 2024

The field of adoptive T cell-based immunotherapy, including tumor-infiltrating lymphocytes (TILs), T cell receptor (TCR) & Chimeric antigen receptor (CAR) - T cells, has undergone unprecedented growth, specifically later in delivering effective and durable clinical response. Notable success with CAR-T cells is limited to hematological malignancies while clinical gains with solid tumors is still a challenge. Solid tumors’ heterogenous target expression and loss, T cell status and metabolic fitness, T cell trafficking into the tumor, and the hostile immunosuppressive tumor microenvironment (TME) have hampered T cell efficacy in the patients. There is a lack of clinically relevant solid tumor models for the translation of T cell-based therapies into the clinic.

We report the development of translational ex-vivo and in-vivo platforms for evaluating cell line-derived and patient-derived xenografts (CDX and PDX) models (BarneyOI™️ model), validated with hCD3 T-cells, autologous TILs and/or allogeneic 3rd generation CAR-T cells (scFv(HER2)-CD28-4-1BB-CD3zeta CAR-T).

Anti-hPDL1 but not anti-hPD1 significantly enhanced hCD3 T-cell and autologous TIL-mediated killing of tumor cells in an ex-vivo co-culture assay using Incucyte-based live cell imaging. Also, a dose-dependent killing of HER-2 expressing CDX (A549 & Colo205) and PDX was observed with HER-2 targeting CAR-T cells in an ex-vivo co-culture assay. This was in concordance with the activation, effector function, and differentiation status of the CAR-T cells, immune phenotype by Cytek aurora spectral flow cytometry. Further, to imitate the patient population and the effect of solid TME, HER-2 targeting CAR-T cells were assessed for their in-vivo efficacy and immune response differences in the subcutaneous and orthotopically implanted HER-2 expressing PDX in immunodeficient NOG mice. Our findings highlight the importance of testing T cell-based immune therapies in the appropriate translational tumor models with better certainty of clinical outcome.

Citation Format: Sherry Liu, Jasmine Borroel, Carmen Sunico, Emily Eastwood, Bridget Corcoran, Bianca Carapia, Jantzen Sperry, Jonathan Nakashima, Rajeev Shrimali. Evaluation of autologous and allogeneic T cell-based therapies in ex-vivo and in-vivo xenograft tumor 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 5342.
Abstract 5394: Nicotine-induced exosomal miR-3157-3p from prostate cancer cells promotes angiogenesis
Authors: Singh Shubhangi , Khan Mohammad Aslam , Anand Shashi , Saranyutanon Sirin , Srivastava Sanjeev Kumar , Singh Seema , Singh Ajay P. , Vikramdeo Kunwar Somesh
Published: March 2024

Nicotine is a psychoactive component of cigarette smoke responsible for addictive properties of tobacco and is shown to promote cancer pathogenesis. We earlier reported that exosomes released from nicotine-treated prostate cancer cells played a key role in tumor angiogenesis, a process necessary to maintain the supply of nutrients and oxygen to the fast proliferating cancer cells. Here, we examined the molecular mechanisms underlying the exosome-mediated effect of nicotine on angiogenesis. For this, we screened the expression of angiogenesis-related miRNAs in the exosomes derived from LNCaP and C4-2 cells treated with vehicle or nicotine (400 nM). Highest and most differential abundance of miR-3157-3p was found in exosomes derived from nicotine-treated prostate cancer cells. Furthermore, the expression of TIMP-2 and KLF-2, established targets of miR-3157-3p, was also downregulated in endothelial cells transfected with exosomes derived from nicotine-treated prostate cancer cells or miR-3157-3p mimics. Studies are currently underway to establish direct targeting of TIMP-2 and KLF-2 by miR-3157-3p and confirm that their suppression is indeed associated with observed changes in endothelial cell growth, migration and capillary-forming ability.

Citation Format: Shubhangi Singh, Mohammad Aslam Khan, Shashi Anand, Sirin Saranyutanon, Sanjeev Kumar Srivastava, Seema Singh, Ajay P. Singh, Kunwar Somesh Vikramdeo. Nicotine-induced exosomal miR-3157-3p from prostate cancer cells promotes angiogenesis [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 5394.
Abstract 5398: Single-cell transcriptomic analysis of partial endothelial-to-mesenchymal transition in sprouting angiogenesis
Authors: Thapa Kapil , Hatch Christopher J. , Hughes Christopher , Fang Jennifer S.
Published: March 2024

Endothelial-to-mesenchymal transition (EndoMT) is a process of morphological, functional, and molecular shift of endothelial cells (EC) towards a mesenchymal identity. This process is critical for embryonic development, and knockout animals lacking copies of the individual master EndoMT transcription factors – including Slug, Snail, and Twist – exhibit significant cardiovascular defects. We previously showed that while Slug knockout animals have a modest developmental vascular phenotype, there is a striking loss of tumor angiogenesis in these animals. Based on these and other data, we hypothesize that sprouting angiogenesis involves transient acquisition of an intermediate state of the EndoMT program –i.e., partial EndoMT (pEndoMT). To test this hypothesis, we used single cell RNA sequencing at day 2 and day 6 following TGF-β activation to map the transcriptomic changes associated with complete endothelial-to-mesenchymal transition. In parallel, we sequenced EC activated to undergo sprouting angiogenesis in vitro at the same timepoints. We show that angiogenic EC transiently acquire a transcriptomic profile of the pEndoMT state, but then later reacquire the transcriptomic profile of committed EC rather than transition completely into mesenchymal cells. We further use this dataset to identify several candidate genes that appear to be regulated by Slug and associated with the pEndoMT state. Future work will use in vitro and in vivo angiogenesis models to identify how pEndoMT is regulated in angiogenesis, and how this regulation may differ in the presence of tumor stroma.

Citation Format: Kapil Thapa, Christopher J. Hatch, Christopher Hughes, Jennifer S. Fang. Single-cell transcriptomic analysis of partial endothelial-to-mesenchymal transition in sprouting angiogenesis [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 5398.
Abstract 5432: The role of nuclear factorκB-inducing kinase in ovarian cancer stem cells, chemoresistance, and relapse
Authors: Lucht Cassidy C. , Robinson Mikella , Gilbert Samuel F. , Olazaba Omar Lujano , House Carrie D.
Published: March 2024

Ovarian cancer is the most lethal gynecological cancer in the United States, largely due to its high relapse rates. Research suggests these high relapse rates are due to cancer stem-like cells (CSCs), a subpopulation of tumor cells that are resistant to chemotherapy and have enhanced tumor-initiation capacity. Post-treatment, CSCs self-renew to re-establish new tumors, leading to recurrent disease that is now resistant to chemotherapy. Given that over-expression of NF-κB has been linked to CSCs, cancer progression, and chemoresistance in various tumors, including ovarian, studies uncovering the mechanisms of this pathway are needed to develop novel therapeutics for ovarian cancer treatment. The upstream activator of the alternative NF-κB pathway, Nuclear Factor κB-inducing Kinase (NIK), could be an enticing target for eliminating CSCs, however its function in these cells is not established. In our current studies, we show that HGSOC cells grown as CSC-enriching 3D spheroids have increased NIK expression and activity relative to monolayer cultured cells. Moreover, cells expressing CSC markers (CD117+/CD133+), have increased NIK gene expression, further supporting an association between NIK and CSC maintenance. Upon treatment with chemotherapies carboplatin and paclitaxel, NIK gene expression increased in the surviving population, indicating a role in chemoresistance. Knockdown of NIK using siRNA led to changes in alternative NF-kB transcriptional activity and higher sensitivity to carboplatin and paclitaxel relative to vehicle controls. Current studies are evaluating the role of NIK in tumor-initiation capacity, asymmetric division, proliferation, and changes in expression of stemness genes. Lastly, an in vivo relapse model will test the feasibility of using a NIK inhibitor following chemotherapy to prevent relapse and prolong survival. Findings from this work will further define the role of NF-kB pathway components in facilitating CSCs survival and re-establishment of tumors and identify a new potential target for eliminating CSCs in ovarian cancer.

Citation Format: Cassidy C. Lucht, Mikella Robinson, Samuel F. Gilbert, Omar Lujano Olazaba, Carrie D. House. The role of nuclear factorκB-inducing kinase in ovarian cancer stem cells, chemoresistance, and relapse [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 5432.
Abstract 5435: CUEDC2 involved in the inhibitory effect of dandelion on triple-negative breast cancer stem cells
Authors: Deng Xinxin , Jiao Yanna , Hao Huifeng , Guo Zhengwang , Xue Dong , Han Shuyan
Published: March 2024

Background: Cancer stem cells (CSCs) exert a substantial influence on the initiation, progression, and metastasis of triple-negative breast cancer (TNBC). Dandelion has exhibited encouraging outcomes in impeding the proliferation of TNBC cells. Nevertheless, the extent of investigation regarding the efficacy of dandelion in targeting CSCs remains restricted.

Purpose: The aim of this study was to investigate the impact of dandelion extract (DE) on CSCs properties.

Methods: In vitro experiments were conducted using TNBC cell lines. Mice bearing TNBC tumors were established for in vivo investigations. Biochemical analysis and cell biological assay were carried to evaluate the effect of dandelion and the underlying mechanisms.

Results: DE treatment significantly decreased the proportion of ALDH+ cells, mammosphere formation ability, and CSCs-related markers. CUEDC2 is highly expressed in ALDH+ TNBC cells, and its overexpression increased the expression of markers related to CSCs. Conversely, the tumor growth was impeded in CUEDC2 KO mice and the markers related to CSCs also were also reduced. CUEDC2 overexpression reversed the inhibitory effect of DE on TNBC CSCs properties. Mechanistically, CUEDC2 related pathway involved in the inhibitory effect of DE on TNBC CSCs properties.

Conclusion: CUEDC2 played a crucial role in maintaining the stem cell properties of TNBC. DE inhibit the self-renewal of TNBC stem cells by regulating the CUEDC2 related signaling pathway.

Keywords: Dandelion; CUEDC2; Triple-negative breast cancer; Cancer stem cell.

Citation Format: Xinxin Deng, Yanna Jiao, Huifeng Hao, Zhengwang Guo, Dong Xue, Shuyan Han. CUEDC2 involved in the inhibitory effect of dandelion on triple-negative breast cancer stem cells [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 5435.
Abstract 5473: Combined inhibition of MDM2 and PARP lead to a synergistic anti-tumoral response in p53 wild-type rhabdomyosarcoma
Authors: Pons Guillem , Gallo-Oller Gabriel , García-Gilabert Lia , Zarzosa Patricia , Sansa-Girona Júlia , Navarro Natalia , Cabré-Fernández Paula , Segura Miguel F , Toledo Josep Sánchez de , Gallego Soledad , Moreno Lucas , Roma Josep
Published: March 2024

Targeting MDM2-p53 interaction to enhance p53 activity represents an attractive antitumoral strategy for p53 wild-type tumors. However, early-phase clinical trials in hematological and solid cancers have shown limited efficacy of MDM2 inhibitors highlighting the necessity for exploring more effective combinations with additional agents. In this study, we provide results from a 22-drug combination screening demonstrating the therapeutic potential of combining Siremadlin (MDM2 inhibitor) and Olaparib (PARP inhibitor) for treating p53 wild-type rhabdomyosarcoma (RMS). Cell survival, cell death and apoptosis analysis revealed synergistic effects when combining Siremadlin and Olaparib in vitro. Combination of both drugs resulted in a significant increase of p53 activity as evidenced by p53 accumulation and K382 acetylation along with an increased expression of p53 targets. Finally, combination of both drugs resulted in significant reduction of tumor growth and increased overall survival outperforming individual treatments and control groups. In summary, our study demonstrates, for the first time, the synergistic effect of the combination between Siremadlin and Olaparib in the inhibition of p53 wild-type RMS tumor growth in vitro and in vivo. These findings advocate for the exploration of this drug combination in clinical trials and underscore the need for further investigation in tumors sharing similar molecular features.

Citation Format: Guillem Pons, Gabriel Gallo-Oller, Lia García-Gilabert, Patricia Zarzosa, Júlia Sansa-Girona, Natalia Navarro, Paula Cabré-Fernández, Miguel F Segura, Josep Sánchez de Toledo, Soledad Gallego, Lucas Moreno, Josep Roma. Combined inhibition of MDM2 and PARP lead to a synergistic anti-tumoral response in p53 wild-type rhabdomyosarcoma [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 5473.