bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 6747: Engineering M13 filamentous phages to target dendritic cells and elicit anti-tumor immunity
Authors: Au-Yeung Yee Man Betty , Zeng Zheng , Huang Jiandong 
Published: March 2024

Abstract
Filamentous phage is a safe and effective vaccine vector that can elicit robust immune responses by activating Toll-like receptor pathways. However, engineering neoantigen-displaying phages for cancer vaccines using phage display technology has been challenging and yielded inconsistent results.
We developed a dendritic cell (DC)-targeting cancer vaccine platform with M13 filamentous phages using the phage display technology. The engineered phages express DC-targeting peptides (SLS) on p3 sites and spy-catcher proteins on p8 positions, allowing neoantigens and proteins of interest to be linked to the phage vector. We named our platform SCP (SLS-spy catcher phage). Imaging and western blot showed that spy-tagged proteins can be easily attached to SCP by simple mixing. Further studies demonstrated that targeting DCs can enhance antigen presentation and anti-cancer immunity.
We tested SCP in MB49 and B16 mouse models and the serum, spleen and tumor tissues were isolated for further investigation of immune response upon treatment. We found that SCP effectively suppressed tumor growth by inducing systemic anti-tumor humoral and cell-mediated immunity. SCP also showed efficacy in late-stage tumor models. SCP treatment triggered particularly robust local anti-tumor response. Histological and flow cytometry analysis revealed increased infiltration of innate and adaptive immune cells, reduced PDL1 expression, and restricted neovascularization in the tumor microenvironment after intratumoral administration of SCP.
Our study demonstrated that SCP can induce multifactorial modifications in the tumor microenvironment to impede tumor growth. SCP overcomes the limitations of traditional phage display technology, enabling a wide variety of peptides and proteins to be loaded onto the phage vectors. SCP is also cost-effective and scalable as phage production and purification processes are relatively mature. We propose that SCP can serve as a universal platform for cancer immunotherapy, and may offer novel treatment options for patients.
Citation Format: Yee Man Betty Au-Yeung, Zheng Zeng, Jiandong Huang. Engineering M13 filamentous phages to target dendritic cells and elicit anti-tumor immunity [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 6747.
Abstract 6754: A microneedle array patch chemo-immunotherapy for cutaneous squamous cell carcinoma
Authors: Yang Yufan , Zhang Haotong , Carey Cara D. , Zhang Jiying , Balmert Stephen C. , Korkmaz Emrullah , Falo Louis D. 
Published: March 2024

Abstract
Cutaneous squamous cell carcinoma (cSCC) is the second most common human malignancy worldwide. Current therapies, including surgical excision, are associated with significant morbidities and considerable expense. Multiple efforts are now underway to develop novel therapies that are effective, durable, safe, cost-effective, and patient-friendly. Here, we report ongoing efforts to develop a localized dissolvable microneedle array patch (MAP) based therapy for cSCC. Our approach is based on the hypothesis that the delivery of an immunogenic cell death (ICD) inducing chemotherapeutic agent to the tumor microenvironment (TME) of accessible cSCCs could kill tumor cells as a source of antigen and induce a proinflammatory TME to generate a potent patient-specific anti-tumor immune response. Specifically, we evaluated MAP therapy using a UV-induced preclinical cSCC mouse model. We applied MAPs delivering the potent ICD-inducing chemotherapeutic agent, doxorubicin (DOX) to UV-induced tumors. MAPs efficiently delivered DOX to the tumor microenvironment. Compared to traditional needle injection, MAP delivery resulted in sustained high levels of DOX in the TME with minimally systemic exposure. Cell death was evident in MAP-DOX-treated tumors, and treatment resulted in tumor regression, increased survival, and long-term immune protection. Further, we found that tumor regression was associated with an early proinflammatory neutrophilic infiltrate. Taken together, our results suggest that MAP delivery of doxorubicin may be a promising approach to cSCC treatment. and that proinflammatory neutrophils are likely an important component of the early infiltrate that leads to tumor regression.
Citation Format: Yufan Yang, Haotong Zhang, Cara D. Carey, Jiying Zhang, Stephen C. Balmert, Emrullah Korkmaz, Louis D. Falo. A microneedle array patch chemo-immunotherapy for cutaneous 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 6754.
Abstract 6792: Investigating differential expression of Palladin isoform 3 in 2D and 3D pancreatic cancer cell cultures
Authors: Biswas Sonali , Bush Jason A. 
Published: March 2024

Abstract
Palladin (PALLD) is an actin-binding protein widely expressed in mammalian tissues that modulates cytoskeletal dynamics. Palladin is linked to the progression of breast, pancreatic, and renal cancers. There are at least 9 known isoforms of PALLD but isoforms 3 (iso3) and 4 (iso4) appear most significant in cancer cells and associated fibroblasts. Our goal is to understand iso3 in pancreatic cancer (PC) cell lines grown in various culture environments to shed light on the intricate interplay between culture conditions and gene regulation. Cell models were developed as 3D spheroids from hanging drops after optimizing culture conditions by utilizing a combination of inexpensive methylcellulose (MC) and rat tail collagen viscosity ranges across cell seeding densities. The quality of the developed spheroids was assessed using multiple parameters, like diameter, integrity, and circularity to determine ideal spheroid characteristics. Spheroids that formed at a lower MC viscosity range (350-550 mPa·s) led to peripheral loose cells and a higher viscosity range (7,000-10,000 mPa·s) resulted in elongated shapes. Moderate viscosity ranges (1,000-1,800 mPa·s and 3,500-5,600 mPa·s) consistently produced circular and compact spheroids. Optimal incubation times varied from 72 to 96 hr depending on the cell line. Invasion dynamics within collagen-MC gels revealed that PANC-1 had significantly higher invasion potential compared to BxPC-3 and a control cell line C2C12. RT-qPCR results show that PALLD iso3 was particularly overexpressed in the 3D culture environment of PANC-1, suggesting that elevated PALLD isoform 3 expression may be a characteristic of 3D culture, particularly in invasive PC types. Taken together, the study validates the importance of extracellular viscosity for influencing spheroid characteristics toward generating reproducible 3D spheroids. The investigation also highlights PALLD iso3 as a potentially valuable biomarker for aggressive forms of pancreatic cancer.
Citation Format: Sonali Biswas, Jason A. Bush. Investigating differential expression of Palladin isoform 3 in 2D and 3D pancreatic cancer cell cultures [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 6792.
Abstract 6833: Microenvironment-specific transcriptomic changes in osteosarcoma metastasis
Authors: Gunawardana-Zeigler Shimara , Peng Dan , Cahan Patrick , Loeb David 
Published: March 2024

Abstract
Osteosarcoma (OS) is the most common primary malignancy of bone in children and adolescents. While conventional chemotherapy has improved 5-year survival rates for localized disease to 70%, metastatic disease remains at 20-30%. Failure of modern treatment options to improve survival reflects the failure of agents that appear promising in preclinical models to demonstrate clinical efficacy in human patients. We hypothesize that this is because most preclinical testing is performed on tumors implanted subcutaneously in immune deficient mice. Our previous work has shown that tumors implanted subcutaneously do not metastasize, whereas genetically identical tumors implanted orthotopically (in the tibia) metastasize to lungs and bones. Thus, to better understand non-genetic drivers of metastasis, we implanted patient-derived xenografts (PDXs) either orthotopically or subcutaneously into NSG mice. Once tumors reached 1.5cm, tumors were extracted along with distant metastases and analyzed by single-cell RNA sequencing. Transcriptomic analysis reveals 1) that fragments of the same tumor implanted subcutaneously have a vastly different transcriptome than fragments implanted orthotopically, 2) the transcriptome of distant metastases is dramatically different from the transcriptome of the primary tumor, and 3) the gene expression profile of metastases is different in different metastatic sites. In all 3 of these circumstances, these differences reflect differences in differentiation status, signaling pathway activation, and immune cell infiltrate. Thus, we conclude that the environment in which a tumor grows significantly alters its gene expression and phenotype. Our study highlights the need for careful consideration of the tumor microenvironment when designing pre-clinical studies to ensure accurate representation of human osteosarcoma and may help to explain the dismal record of translating novel agents from preclinical studies into standard therapy.
Citation Format: Shimara Gunawardana-Zeigler, Dan Peng, Patrick Cahan, David Loeb. Microenvironment-specific transcriptomic changes in osteosarcoma 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 6833.
Abstract 6901: Paired colorectal and pancreatic cancer patient-derived tumor and organoids biobank
Authors: Wang Juiling , Ma Wen-Hui , Lo Yun-Chi , Hsieh Hsiao-Chun , Hsiao Li-Ju , Bau Yu-Pin , Yang Nai-Chieh , Tsai Shaw-Jenq , Shan Yan-Shen , Chen Tzu-Yu , Chen Yu-Ling , Chiang Pei-Ju , Chen Tzu-Ling , Chang Hsiu-Ping , Chin Hsian-Jean 
Published: March 2024

Abstract
Patient-derived xenograft (PDX) models constitute a crucial tool in precision cancer medicine owing to their high accuracy in predicting the response to therapeutic drugs among patients in clinical settings. Our center has established a biobank that collects more than 80 colorectal cancer and 25 pancreatic cancer PDX tissues, along with essential information about patients’ tumors and PDX models. However, cost-intensive and time-consuming limit the usage of PDX model in high throughput screening during drug development. To overcome this hurdle, we are generating a PDX-match paired organoid (PDXO) biobank by using emerging three-dimensional culture techniques. Before large-scaled expansion, the concordance between PDX tissues and PDXOs of partial specimens was evaluated firstly using short-tandem repeat (STR) analysis, immunohistochemistry (IHC) and H&E staining. Data of STR analysis performed by using AmpFLSTR Identifiler PLUS PCR Amplification Kit demonstrated that PDX tissues and PDXOs have identical DNA fingerprinting. IHC and H&E staining revealed PDX tissues and PDXOs have highly similar organ-specific markers expression (e.g., Cytokeratin 20, Caudal Type Homeobox 2, and Pan Cytokeratin) and histological morphology. Even after passing to at least five generations, passed PDXOs maintained high fidelity to their original state, as confirmed by whole-exome sequencing (WES) and STR analysis across passages. The WES analysis for passage 0 (P0) and passage 5 (P5) of PDXOs from different PDX tissues revealed there is more than 95% intersection between P0 and P5 PDXOs with the same missense mutation gene. The results of STR analysis showed that P0 and P5 PDXOs have identical STR profile. To evaluate the capability of PDXOs in clinical response prediction, several effective and ineffective drugs (e.g., oxapliplatin, 5-FU, gemcitabine) were tested. The results of drug testing indicated that PDXOs can be used to accurately predict the response to therapeutic drugs as PDX model but more efficient than PDX model. Therefore, PDXO biobank can serve as a valuable resource for high throughput in vitro drug screening prior in vivo efficacy confirmation using PDX models.
Citation Format: Juiling Wang, Wen-Hui Ma, Yun-Chi Lo, Hsiao-Chun Hsieh, Li-Ju Hsiao, Yu-Pin Bau, Nai-Chieh Yang, Shaw-Jenq Tsai, Yan-Shen Shan, Tzu-Yu Chen, Yu-Ling Chen, Pei-Ju Chiang, Tzu-Ling Chen, Hsiu-Ping Chang, Hsian-Jean Chin. Paired colorectal and pancreatic cancer patient-derived tumor and organoids biobank [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 6901.
Abstract 6958: CRIPTO-regulated extracellular vesicles mediate heterotypic cellular crosstalk in triple negative breast cancers
Authors: Freeman David W. , Gates Brooke L. , Spendlove Mauri D. , Spike Benjamin T. 
Published: March 2024

Abstract
Tumor progression involves dynamic interaction between various cell types within the tumor microenvironment. Fibroblasts, which normally participate in wound healing, reciprocally communicate with breast cancer cells to promote tumor cell plasticity and heterogeneity. Gaining insights into the molecular mechanisms underlying this cell-to-cell communication, could unlock novel therapeutic avenues for inhibiting cellular plasticity, breast cancer progression and metastasis. The small GPI-anchored adaptor protein CRIPTO is a key player in this context though important molecular details remain to be elucidated. CRIPTO orchestrates TGF-β pathway signaling as well as growth factor-like signaling pathways that activate c-Src/MEK/AKT, in an autocrine and paracrine manner. CRIPTO is expressed during mammary development, wound healing, and breast cancer but is largely undetectable in homeostatic tissue, making it a promising therapeutic target. Our previous studies have demonstrated that the activity of CRIPTO is contingent upon cellular stress, with pronounced influence within pro-fibrotic, hypoxic domains of tumors. Importantly, we have shown that inhibition of CRIPTO using the soluble recombinant protein antagonist A4Fc effectively impedes the progression of triple negative breast cancer (TNBC) in murine xenograft models. Building upon these findings, we have been dissecting CRIPTO’s role in coordinating fibroblast activation and cell-state reprogramming in TNBC. We find that inhibition of CRIPTO with A4Fc suppresses cancer associated fibroblast (CAF) activation and collagen remodeling in vivo. Further, by modeling tumor cell:fibroblast crosstalk in vitro, we have demonstrated mechanistically that CRIPTO expression in tumor cells mediates subsequent extracellular vesicle (EV) uptake by fibroblasts, as well as fibroblast SMAD pathway activation and priming for reciprocal signaling back to tumor cells to promote transcriptional reprogramming and cellular invasion.The significance of these findings lies in their elucidation of key EV control mechanisms that could be exploited to intervene in essential tumor cell:fibroblast crosstalk or to enhance targeting of therapeutic moieties based on EV architecture and function in cancer and/or normative regenerative processes.
Citation Format: David W. Freeman, Brooke L. Gates, Mauri D. Spendlove, Benjamin T. Spike. CRIPTO-regulated extracellular vesicles mediate heterotypic cellular crosstalk in triple negative breast cancers [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 6958.
Abstract 6959: Acetylcholine potentiates invasion and migration of human pancreatic cancer cells under hypoxia through α7 nAChR-mediated increases in HIF-1α expression
Authors: Cho Yunmi , Oh Eun-Taex , Kim Ha Gyeong , Park Heon Joo 
Published: March 2024

Abstract
The most significant nerve-related problem in clinical pancreatic cancer management is perineural invasion (PNI), characterized as the neoplastic invasion of cancer cells into or surrounding the nerves. It is a characteristic feature of pancreatic ductal adenocarcinoma (PDAC) associated with a poor prognosis, tumor recurrence, and emergent pain. However, why PDAC is associated with a higher incidence of PNI remains unclear. HIF-1 (hypoxia-inducible factor-1) regulates the expression of ~1000 genes involved in angiogenesis, metastasis, tumor growth, chemoresistance and radioresistance, underscoring the growing interest in targeting HIF-1 for cancer control. In the present study, we investigated the molecular mechanisms underlying neurotransmitter acetylcholine (Ach)-potentiated HIF-1α expression and subsequent increase in invasion and migration of human pancreatic cancer cells in hypoxia (2% O2). The expression of various acetylcholine receptors in six types of pancreatic cancer cell lines (MIA-PaCa-2, PANC-1, HPAF-II, Capan-1, AsPC-1, and BxPC-3 cells) was observed using qPCR, and α7 nAChRs was observed to be expressed in all pancreatic cancer cells. Therefore, we chose MIA-PaCa-2 cells, expressing high levels of α7 nAChRs, and HPAF-II cells, expressing low levels of α7 nAChRs, for loss- and gain-of-function studies, respectively. In vitro assays showed that, under hypoxic conditions, Ach caused a concentration-dependent increase in α7 nAChRs-mediated HIF-1α expression, enhancing expression of HIF-1α target genes, and promoting invasion and migration of human pancreatic cancer cells. We further found that Ach increased HIF-1α protein stability in pancreatic cancer cells under hypoxic conditions. This effect was mediated by YAP signaling, as evidenced by the fact that Ach-induced increases in HIF-1α protein stability were blocked by siRNA-mediated knockdown of YAP. Consistent with these findings, an investigation of the relationship between α7 nAChRs and clinicopathological features in pancreatic cancer patients using publicly available datasets showed that high levels of α7 nAChRs are correlated with a poorer prognosis in pancreatic cancer patients. Collectively, these findings identify novel mechanisms by which Ach potentiates invasion and migration of pancreatic cancer cells and provide further evidence that HIF-1α is a potential anticancer target in Ach-associated pancreatic cancer.
Citation Format: Yunmi Cho, Eun-Taex Oh, Ha Gyeong Kim, Heon Joo Park. Acetylcholine potentiates invasion and migration of human pancreatic cancer cells under hypoxia through α7 nAChR-mediated increases in HIF-1α expression [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 6959.
Abstract 6965: Characterizing the proteome landscape of extracellular vesicles from prostate cancer cell lines by mass spectrometry
Authors: Ludwig Megan L. , Day Abderrahman , Bergom Hannah E. , Sychev Zoi , Horrmann Alec , Hwang Justin , Drake Justin M. 
Published: March 2024

Abstract
Multiple subtypes of prostate cancer can emerge in advanced stages of disease, such as tumors that maintain androgen receptor features (AR+), those without AR but with neuroendocrine features (AR-/NE+), and those without AR or NE features (AR-/NE-). These latter subtypes are particularly aggressive and incredibly lethal. Extracellular vesicles (EVs) are small particles that are mediators of cell-cell communication and have been nominated as potential biomarkers of disease state. We hypothesized that the proteome of EVs contains information about their originating cell that can distinguish between prostate cancer subtypes. To address this, we isolated EVs from cell lines of differing subtypes – AR+ (LNCaP, LNCaP95, C4-2, 22RV1), AR-/NE+(NCI-H660, EF1, LASCPC-01), and AR-/NE- (PC3, DU145) – and prepared the EV proteome for quantitative, label-free mass spectrometry analysis. This identified 2,000+ proteins in the EVs for each cell line where EVs from similar subtypes clustered together. Interestingly, we also found that EVs contained enough protein information to utilize developed gene signatures for AR or NE activity. EVs isolated from AR-/NE+ cells contained known NE signature proteins such as SYP, CHGA, and UCHL1. EVs from the AR+ cell lines displayed &gt;50% AR activity driven by PSA and STEAP1 proteins in addition to other significant distinguishing protein markers. This supports the potential for using the proteome of tumor-derived EVs to identify novel signature molecular markers besides those already known to the field for developing liquid-based biomarker assays. In addition, our work characterizes the EV surfaceome which is critical for enriching signals from tumor-derived EVs feasible for clinical diagnostics assay development. While PSMA has been a reliable marker for cells reliant on AR signaling, less is known about protein markers in the AR- subset. Our work highlights TROP2 as especially enriched in the EVs from AR-/NE- cell lines, supporting its relevance as a target in patients without treatment options. To evaluate disease outcomes, further clinical studies are needed to develop specific and sensitive tests based on the EV proteome derived from advanced prostate cancer patients.
Citation Format: Megan L. Ludwig, Abderrahman Day, Hannah E. Bergom, Zoi Sychev, Alec Horrmann, Justin Hwang, Justin M. Drake. Characterizing the proteome landscape of extracellular vesicles from prostate cancer cell lines by mass spectrometry [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 6965.