bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 6740: Next-generation LNPs induce effective anti-tumor T cell responses
Authors: Millar Douglas G. , Gold Matthew J. , Olsen Kirsten , Wu Yu , Karpov Yury , Nechanitzky Robert , Menon Haritha , Krishnan Rajesh , Georgantas Robert , Ohashi Pamela , Martin-Orozco Natalia 
Published: March 2024

Abstract
mRNA immunotherapies have gained interest for the treatment of tumors from various origins, however, their ability to provide long -term durable tumor control has been underwhelming. Here we report that novel mRNA lipid nanoparticle (LNP) vaccinations induce robust expansion of antigen-specific T cells, although this does not always correlate with their ability to infiltrate and kill solid tumors. Using a portfolio of novel ionizable lipids and a robust in vivo model that reliably identifies LNPs with prominent anti-tissue T cell activity we have identified a putative LNP candidate that provides long term tumor control in a murine syngeneic tumor model.
We utilized the model antigen glycoprotein (Gp), from the lymphocytic choriomeningitis virus (LCMV), and produced mRNA encoding the immunodominant epitopes gp3333-41, gp6161-80 and gp276276-286 (3GP-mRNA). This mRNA was encapsulated in various novel LNP formulations and administered i.m. into naïve C57Bl/6 mice to track the induction of a robust CD8+ T cell response identified using tetramers to gp33 and gp276. Successful candidate LNPs were then tested in the RIPgp model of autoimmune diabetes. RIPgp transgenic mice express LCMV-Gp in the pancreatic β cells and offer a robust assay for the ability to induce antigen-specific T cells capable of breaking self-tolerance and infiltrating and killing target tissue. Utilizing the RIPgp model to screen LNP candidates we identified a lead candidate, C2-5A, that both induced a robust expansion of antigen-specific T cells and tissue destruction. When utilized in a colon adenocarcinoma model expressing the LCMV-Gp antigen (MC38-Gp), therapeutic dosing of LNP C2-5A as a monotherapy lead to long term tumor clearance in 40% of the treated animals. We believe this screening platform allows the selection of LNP candidates capable of breaking self-tolerance, and one such candidate C2-5A provides a potent vehicle and adjuvant for mRNA based cancer vaccines.
Citation Format: Douglas G. Millar, Matthew J. Gold, Kirsten Olsen, Yu Wu, Yury Karpov, Robert Nechanitzky, Haritha Menon, Rajesh Krishnan, Robert Georgantas, Pamela Ohashi, Natalia Martin-Orozco. Next-generation LNPs induce effective anti-tumor T cell responses [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 6740.
Abstract 6758: Eradication of tumors and development of immunity from pHLIP-targeted intracellular delivery of STINGa
Authors: DuPont Michael , Moshnikova Anna , Iraca Marissa , Klumpp Craig , Visca Hannah , Allababidi Dana , Pelzer Phoebe , Engelman Donald , Andreev Oleg , Reshetnyak Yana 
Published: March 2024

Abstract
Activation of the stimulator of interferon genes (STING) pathway causes the release of factors that trigger the immune response in the tumor microenvironment (TME). Transient strong activation leads to immune activation and tumor suppression, while prolonged weak activation leads to immuno-suppression promoting tumor development. Reprogramming of M2-type tumor-associated macrophages (M2-TAMs) toward an M1 phenotype, suppression of cancer-associated fibroblasts (CAFs) and myeloid-derived suppressor cells (mMDSCs), as well as activation of dendritic cells (DCs) to train T-cells are advantageous, while the activation of STING pathway in T-cells is leads to their apoptosis. Thus, targeted delivery of STINGa to specific cells with TME to induce strong transient activation may be a key to therapy. We have developed a delivery approach that uses one or two pH Low Insertion Peptides (pHLIPs) that collaborate in the targeted intracellular delivery of a STINGa. STINGa were conjugated with pHLIPs via S-S cleavable self-immolating linkers. Biophysical studies were performed to ensure proper interactions of pHLIP-STINGa agents with membrane lipid bilayers. pHLIPs extend the lifetime of STINGa in the blood and target them to acidic cancer and stromal cells (CAFs), as well as M2-TAMs, mMDSCs and DCs within TME. The targeting results in selective cytokine activation in tumors with no systemic activation, which triggers the eradication of small (100 mm3) and large (400-700 mm3) CT26 tumors in mice after one or two doses of pHLIP-STINGa. The tumor stroma was destroyed, intratumoral hemorrhage developed, and the level of acidity within the TME was reduced. No tumors developed in mice re-challenged by an additional injection of cancer cells in a 90-day experiment. Thus, targeted delivery of STINGa to cancer cells, tumor stroma and TAMs induces activation of signaling, potentially resulting in the recruitment and infiltration of T- and NK-cells, which gain access to the tumor core. The cytotoxic activity of T- and NK-cells is not impaired by the acidic environment and immune memory is developed. The pHLIP technology may allow transformation of immuno-activating agents into more potent therapeutics, since pHLIP can target and deliver these agents to cancer cells, tumor stroma and myeloid cells.
Citation Format: Michael DuPont, Anna Moshnikova, Marissa Iraca, Craig Klumpp, Hannah Visca, Dana Allababidi, Phoebe Pelzer, Donald Engelman, Oleg Andreev, Yana Reshetnyak. Eradication of tumors and development of immunity from pHLIP-targeted intracellular delivery of STINGa [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 6758.
Abstract 6778: New microplate platform with cryopreserved 3D tumor spheroids for faster and convenient pre-clinical testing
Authors: Goedecke Nils , Stroebel Simon , Hutter Simon , Issa Tala , Agarkova Irina , Ligeon Laure-Anne , Frey Olivier 
Published: March 2024

Abstract
Drug discovery and safety testing depend on predictive in vitro models, including multicellular tissues with organotypic functions. While traditional 2D tumor cell cultures have been extensively used in early drug screening due to their user-friendly nature, automation compatibility, and high throughput, they fall short in representing the intricate 3-dimensional structure, physiology, and complex multicellular nature of actual tumors. However, generating such 3D tissues for each test campaign is labor intensive. In addition, not all researchers have the resources to establish these 3D models. To pave the way, we developed a new platform that offers a range of cryopreserved 3D tumor models in micro plate format allowing for high-content screening. It enables users to start testing compounds within a 3-to-5 days simply by thawing an ARCTis™ plate (Always Ready Cryo Tissues) and following clear instructions. Another advantage of these cryopreserved 3D models derives from the fact that they are made in larger batches which means the start point for compound testing is always the same. This improves data quality, maximizes the test window length, and makes the data comparable across work groups. Currently, our growing portfolio consists of 25 tumor cell lines from ATCC representing the most common solid tumors. Their functionality has been determined based on efficacy tests for each cell line with the apoptotic agent Staurosporine. Various cell lines exhibited compact spheroids with a round morphology, less compact aggregates with non-spherical morphology, or loose cell-cell interaction aggregates, which can correlate with their tumor site of origin. Upon treatment with Staurosporine the dose-response profiles indicated growth inhibition, tumor shrinkage, or tumor cell killing effects based on size assessment and cell metabolic analysis (ATP). Moreover, we demonstrated the robust performance of these assays from plates stored for up to 6 months at -80°C.In conclusion, this groundbreaking approach addresses the limitations of traditional 2D cell cultures by providing a high-throughput, in-plate frozen, 3D cell culture platform. By offering a combination of a more physiologically relevant representation of tumors with a robust frozen-storage and time flexible assay platform our method holds great promise for advancing cancer drug discovery by easing the exploration of complex drug testing screens in vitro.
Citation Format: Nils Goedecke, Simon Stroebel, Simon Hutter, Tala Issa, Irina Agarkova, Laure-Anne Ligeon, Olivier Frey. New microplate platform with cryopreserved 3D tumor spheroids for faster and convenient pre-clinical testing [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 6778.
Abstract 6800: The tumor progression mechanism of increasing CXCL16-CXCR6 pathway within the tumor microenvironment in TNBC
Authors: Lee Yun sun , Lee Cham Han , Ha Seong Yun , Lee Sang Hyeob , Yang Jin Yi , Kim Young Sang , Kim Sung Moo , Kim Seong Keun 
Published: March 2024

Abstract
Background: Triple negative breast cancer (TNBC) is characterized rapid progression, higher relapse, and poor prognosis, so the establishment of an effective therapeutic target is required. CXCL16, a chemokine abundantly present in the TNBC tumor microenvironment (TME), is secreted by tumor-associated macrophages (TAM) and is known to promote tumor growth and metastasis. However, the exact mechanism through which CXCL16 facilitates tumor growth and metastasis remains unclear. This study aims to study the role of CXCL16 in the tumor progression and confirm the anti-tumor effects of CXCL16 neutralization in TNBC model.
Method: MDA-MB-231 human TNBC cell lines were utilized for in vitro validation, and a syngeneic model involving 4T1 mouse TNBC cell lines and Balb/c mice was used for in vivo studies.
Results: Stimulation of TNBC cells with recombinant human CXCL16 protein (rhCXCL16) activates the tumor growth signaling pathways, including pAKT and STAT3, as well as EMT markers. These results shows that CXCL16 can promote tumor growth and metastasis. Treatment with CXCL16 neutralizing antibodies inhibited the promotion of cancer growth and metastasis by rhCXCL16. Additionally, combination treatment of CXCL16 antibodies with PD-1 antibodies in the 4T1 syngeneic model increased the proportion of CD8+ T cells and CD80+ M1 like macrophages. These results shows that neutralization of CXCL16 can regulate polarization of macrophage and proportion of T cells. These regulation and changes of immune cells allow neutralization of CXCL16 control the TME to resist tumor cells, suggesting that optimal condition can be constructed to maximize the effectiveness of anti-cancer drugs.
Conclusion: This study confirms that the CXCL16-CXCR6 pathway within the TME triggers tumor growth and metastasis, while neutralization of CXCL16 can inhibit tumor progression. This effect is attributed to the increase in CD8+ T cell and M1 macrophage proportions within the TME. In conclusion, these research findings propose the potential of CXCL16 as a novel biomarker and therapeutic target for TNBC.
Citation Format: Yun sun Lee, Cham Han Lee, Seong Yun Ha, Sang Hyeob Lee, Jin Yi Yang, Young Sang Kim, Sung Moo Kim, Seong Keun Kim. The tumor progression mechanism of increasing CXCL16-CXCR6 pathway within the tumor microenvironment in TNBC [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 6800.
Abstract 6879: Development of a rapid multiplex immunohistochemistry for characterizing tumor-immune microenvironment
Authors: Kimura Alisa , Tsujikawa Takahiro , Morimoto Hiroki , Mitsuda Junichi , Saburi Sumiyo , Mukudai Shigeyuki , Nagao Hikaru , Sakurai Nana , Murakami Nanako , Miyagawa-Hayashino Aya , Ogi Hiroshi , Shibata Saya , Konishi Eiichi , Itoh Kyoko , Hirano Shigeru 
Published: March 2024

Abstract
Intratumoral immune profiles have been related to prognosis and therapeutic efficacy; this could result in personalized treatment based on biomarkers. Although techniques for multifactorial evaluation of different tumor-immune microenvironments have been established, their practical implementation in clinical settings still requires further improvement, where the time required for testing needs to be reduced and the technology needs to be clinically validated. To develop a multiplex, quantitative, and rapid tissue evaluation method based on clinically established standard immunohistochemistry (IHC), we attempted to develop 6-marker rapid multiplex IHC based on our previously reported 14-marker multiplex IHC via the reduction of the number of labels and speeding up the staining procedure (Tsujikawa et al. Cell Reports, 2017). First, we confirmed whether immunological features that were found to be associated with prognosis in 14-marker multiplex IHC analyses could be identified with fewer labels. Quantitative results by selected six markers exhibited significant correlation with those by 14 markers in terms of immune classifications based on the balance of lymphoid and myeloid cells. Next, we developed a rapid staining protocol by optimizing the reaction temperature, chromogen, and washing time, allowing the completion of 6-marker analysis in 5 hours and 49 minutes as opposed to several days required for the conventional multiplex IHC. By using six markers, validation in benign tonsil tissue and head and neck cancer tissue exhibited significant correlation between rapid and standard multiplex IHC in terms of staining intensities, densities of T cells, macrophages, lymphoid/myeloid immune cells ratios, and spatial profiles of intratumoral immune infiltrates. This method may enable a quantitative assessment of tumor-immune microenvironment on a clinically feasible time scale, which promote the development of tissue biomarker-guided therapeutic strategies.
Citation Format: Alisa Kimura, Takahiro Tsujikawa, Hiroki Morimoto, Junichi Mitsuda, Sumiyo Saburi, Shigeyuki Mukudai, Hikaru Nagao, Nana Sakurai, Nanako Murakami, Aya Miyagawa-Hayashino, Hiroshi Ogi, Saya Shibata, Eiichi Konishi, Kyoko Itoh, Shigeru Hirano. Development of a rapid multiplex immunohistochemistry for characterizing tumor-immune microenvironment [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 6879.
Abstract 6885: Investigating a novel role of DCLK1 isoforms in colitis and colitis associated colon cancer
Authors: Yusuf Kafayat Aderonke , Roy Badal C. , Anant Shrikant , Umar Shahid 
Published: March 2024

Abstract
Chronic intestinal inflammation in patients diagnosed with inflammatory bowel disease (IBD) significantly increases their risk of colorectal cancer (CRC). Despite the rising global frequency of IBD, the precise mechanisms that regulate the transition from IBD to colon cancer are poorly understood. Doublecortin-like kinase 1 (DCLK1) is a protein with two major isoforms, DCLK1-L and DCLK1-S. Previous studies have shown that hypermethylation of the DCLK-1α promoter encoding DCLK1-L, particularly in colorectal cancer, allows switching to the DCLK1-S isoform that confers an invasive tumor phenotype. The DCLK1-L is a tuft cell marker that protects against colitis by sensing and eliminating gut pathogenic infection through various mechanisms. On the contrary, the role of DCLK1-S in colitis and colitis-associated cancer has not been thoroughly investigated. Studies from our lab have identified a marked elevation of DCLK1-S in tissue samples of IBD patients. We also recently discovered that DCLK1-S is predominantly expressed in Ly6G+; MHCII- neutrophils, which coincide with elevated levels of inflammation and tissue damage in the colon. To further understand the role of DCLK1 isoforms in driving the progression of colitis to colon cancer, we bred Dclk1fl/fl mice with MRP8-Cre-ires/GFP mice to generate Dclk1fl/fl;MRP8-Cre+/- that eliminates DCLK1-L upon tamoxifen injection resulting in sustained expression of DCLK1-S in the granulocytes especially neutrophils. When infected with Citrobacter rodentium (CR), MRP8;Dclk1−/− mice compared to MRP8;Dclk1+/− or WT mice developed more severe colitis. We have also discovered a novel interaction between DCLK1-S and MMP13 in an unbiased docking study that correlated with significant co-localization of DCLK1-S with MMP13 in the colons of CR-infected mice. Employing a knock-in mice over-expressing the DCLK1-S +19 under the CAG promoter, the DCLK1-MMP13 co-localization correlated with severe ulceration and colitis with loss of crypts and immune cell infiltration including Ly6G+ neutrophils. Studies are underway to see if the severity of colitis in MRP8;Dclk1−/− mice translates into enhanced tumorigenesis and to see if potential interaction between DCLK1-S and MMP13 could provide a plausible mechanism of colitis progression to colon cancer through ECM remodeling and tissue damage.
Citation Format: Kafayat Aderonke Yusuf, Badal C. Roy, Shrikant Anant, Shahid Umar. Investigating a novel role of DCLK1 isoforms in colitis and colitis associated colon 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 6885.
Abstract 6905: A multi-targeted approach to evaluate therapeutic selection and efficacy in preclinical GBM models
Authors: Eastwood Emily , Carapia Bianca , Rodriguez Javier , Buck Kristen , Gorospe Derrick , Valencia Elizabeth , Corcoran Bridget , Pippa Raffaella , Chien Yuan-Hung , Andrews Warren , Do Long , Nakashima Jonathan K. , Sperry Jantzen 
Published: March 2024

Abstract
Glioblastoma (GBM) is a devastating primary brain cancer with approximately 10,000 new US diagnoses annually. The current standard of care (SOC) for GBM includes surgical resection followed by radiation therapy (RT) and temozolomide (TMZ), however, there is near universal recurrence and development of resistance after treatment. Relapse in disease is tightly linked with dynamic changes in gene expression during tumor evolution, highlighting the need for stronger preclinical GBM models. Here, we report developing a pair of patient-derived xenograft (PDX) models from surgical resections obtained from initial GBM incidence in a patient (BarneyOI™ Cancer Model CRT00433) and subsequent recurrence following treatment (BarneyOI Cancer Model CRT00435) to study disease progression and novel treatment strategies. Recent studies have suggested the utilization of combination therapy approaches for GBM patients to address TMZ resistance. To this end, Certis has developed a personalized, AI-based approach to predict and test combination therapies in preclinical cancer models. Accuracy of the predicted sensitivities to treatment was evaluated in vivo using both subcutaneous (SC) and orthotopic (OT) mouse models. Luciferase-tagged CRT00433 and CRT00435 spheroid cell lines were transduced with firefly luciferase and implanted intracranially by stereotactic surgery for OT monitoring. Optical bioluminescence imaging (BLI) and murine-scale MRI from Aspect Imaging were used to assess therapeutic response of AI-predicted combination therapies and SOC treatment.
Citation Format: Emily Eastwood, Bianca Carapia, Javier Rodriguez, Kristen Buck, Derrick Gorospe, Elizabeth Valencia, Bridget Corcoran, Raffaella Pippa, Yuan-Hung Chien, Warren Andrews, Long Do, Jonathan K. Nakashima, Jantzen Sperry. A multi-targeted approach to evaluate therapeutic selection and efficacy in preclinical GBM 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 6905.
Abstract 6926: Tracing clonal evolution and immune evasion in glioblastoma progression
Authors: Ceresa Davide , Garcia Mena Sebastian A. , Piaggio Francesca , Irene Appolloni , Marubbi Daniela , Malatesta Paolo 
Published: March 2024

Abstract
Glioblastoma continues to be a daunting obstacle in oncology research, with its early-stage progression being particularly enigmatic. In a recent published work, we traced the clonal dynamics of glioblastoma evolution by the simultaneous transfer of PDGFB and genetic barcodes into mouse brains. We observed a continuous clonal loss during the acquisition of a malignant phenotype, underlined by the modulation of the levels of c-Myc expression and their functional targets. In this study, we delve deeper into the evolution of glioblastoma by transplanting multiclonal, early-stage glioma cells into multiple immunodeficient NOD-SCID mice. This experimental design allowed us to track the clonal dynamics over serial transplants, revealing how early-stage glioma clones acquire immune-evasive features capable of initiating tertiary tumors in immune-competent hosts. Through barcode sequencing and single-cell RNA sequencing of early-stage gliomas, coupled with bulk RNA sequencing of secondary and tertiary gliomas, we dissected the clonal and transcriptomic landscape of these tumors. We observed that, of the various clones composing primary tumors, just a few of them are predominating in subsequent passages. Furthermore, the clonal composition of secondary tumors derived from the same primary glioma showed partial overlap, indicating a partial predetermination in the development of immune-evasive behavior. Our intra- and inter-clonal transcriptomic analysis across various stages of tumor progression is shedding light on how new functional traits can emerge in gliomas. Further analysis of our data will determine whether it is the result of the expansion of clones already possessing these traits, or if they stem from functional adaptations within the clones themselves. Our insights further reinforce the concept of clonal competition, highlighting the significant influence of immune-system interactions in driving these competitive dynamics.
Citation Format: Davide Ceresa, Sebastian A. Garcia Mena, Francesca Piaggio, Appolloni Irene, Daniela Marubbi, Paolo Malatesta. Tracing clonal evolution and immune evasion in glioblastoma progression [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 6926.