bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2565: Divergentnucleocytoplasmic transport influences escape from HER2-activated DCIS-like state
Authors: Wang Lixin , Paudel Bishal , McKnight Anthony , Janes Kevin 
Published: April 2023

Abstract
HER2/ErbB2 and EGFR/ErbB1 activation incite a phenotype resembling premalignant escape from ductal carcinoma in situ (DCIS) in the 3D multicellular arrangement of breast-epithelial cells. This phenomenon tends to be infrequent, and the exact mechanism for its heterogeneous presentation has been elusive. We sought to investigate the process by which ErbB receptors facilitate a picture of incomplete penetrance in DCIS escape (DE). To identify the transcriptional signatures that prime cells towards DE, we randomly profiled 10-cell transcriptomes of single spheroids after 24 hours of ERBB activation, and frequency matched gene expression to the DE-phenotype. We uncovered a network of nucleocytoplasmic transport (NCT) regulators that alter the DE-phenotype penetrance. Of the regulators, CSE1L (an exportin) synergistically increases the DE-frequency when induced, while its knockdown reduces the phenotype, suggesting its functional role. Furthermore, we evaluated the significance of this result in vivo by perturbing Cse1l in Erbb2-amplified mouse mammary-gland tumors and found that activation of Erbb2 (mimicked by ERBB-inhibitor release) alters the macroscale organization of tumors, reminiscent of DE. Specifically, we found an elevated CSE1L inhibits an accumulation of classical nuclear-localizing cargoes that require binding with importin-α/β (KPNAs, KPNB1) complexes. Using proximity labeling with BirA*-fused with CSE1L, we identified ERBB1/ERBB2/ERBB4 as CSE1L interactors. Mechanistically, hetero-dimerized ERBB receptors interact with importin-α/β complexes and re-localize to the nucleus, corroborating previous observations of receptors’ nuclear translocation. Intriguingly, we observed that Doxycycline-induced knockdown of clathrin heavy chain (CLTC, involved in intracellular trafficking of receptors via endocytosis) synergizes with ERBB activation to increase the DE-phenotype penetrance. Using ChIP-Seq, we found that ERBB1 binds to the locus of microRNA, miR205, which negatively regulates the expression of importin-α, mainly KPNA1. These results indicate that ERBB receptors translocated to the nucleus reduce the DE-phenotype by inhibiting importins via miR205, while CSE1L induction relieves this inhibition by reducing the receptor's nuclear accumulation. Taken together, these results show that nucleocytoplasmic shuttling of receptors in ERBB-active breast epithelia generates tunable cross-inhibitory feedback through nucleocytoplasmic transport that results in long-term, heterogeneous multicellular fates.
Citation Format: Lixin Wang, Bishal Paudel, Anthony McKnight, Kevin Janes. Divergentnucleocytoplasmic transport influences escape from HER2-activated DCIS-like state [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2565.
Abstract 2566: Blocking the functional domain of TIP1 upregulates midkine via the β-catenin/Wnt signaling pathway
Authors: Saikia Minakshi , Singh Abhay K. , Cheung Nathan , Hallahan Dennis E. , Kapoor Vaishali 
Published: April 2023

Abstract
Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related mortalities world-wide. Targeted therapy has been a growing topic of investigation to improve therapeutic efficacy for NSCLC. A few targeted therapies that exploit aberrant protein expression profiles have been approved for NSCLC. However, marginal improvement in efficacy observed with these therapeutic approaches highlights the need for discovery of additional therapeutic targets. Tax interacting protein 1 (TIP1) is a molecular target that is overexpressed on the surface of various cancers and correlates with poor overall patient survival. The functional domain of TIP1 (PDZ domain) caps the C-terminus of many cellular proteins that regulate cell signaling, cancer development, and progression. We found that antibodies against the PDZ domain of TIP1 induced cytotoxicity of lung cancer cells but not normal cells. A quantitative mass spectrometry-based approach identified a ten-fold increase in Midkine protein expression on blockade of TIP1. Midkine is a heparin-binding growth factor that is involved in cancer progression, therapy resistance, and immune suppression. This study tested the hypothesis that Midkine is upregulated by the anti-PDZ/TIP1 antibody via the β-catenin/Wnt signaling pathway, which subsequently modulates downstream signaling mechanisms. In co-immunoprecipitation studies we found a direct interaction of TIP1 with β-catenin on the cell surface. Anti-TIP1 antibody treatment led to nuclear translocation of β-catenin in NSCLC cells. We used a luciferase based TOPFlash/FOPFlash reporter assay to study the transcriptional activity of the canonical β-catenin/Wnt pathway following TIP1 blockade. Reporter assays showed a three to four-fold increase (p&lt;0.0001) in the transcriptional activity of β-catenin following TIP1 blockade in NSCLC cells. Genetic silencing of β-catenin inhibited Midkine mRNA transcription (p&lt;0.005), protein induction and its extracellular release (p&lt;0.0001) following anti-TIP1 antibody treatment. In conclusion, the mechanism of Midkine release following TIP1 blockade involves the β-catenin/Wnt signaling pathway. Ongoing studies are evaluating the dual-targeting of TIP1 and Midkine for enhanced efficacy of NSCLC treatment.
Citation Format: Minakshi Saikia, Abhay K. Singh, Nathan Cheung, Dennis E. Hallahan, Vaishali Kapoor. Blocking the functional domain of TIP1 upregulates midkine via the β-catenin/Wnt signaling pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2566.
Abstract 2567: Exploring novel roles of the GPCR Smoothened in AKT signaling and drug resistance
Authors: Manuel Samantha , Agarwal Nitin , Bilbao Daniel , Vega Francisco , Landgraf Ralf 
Published: April 2023

Abstract
The Smoothened receptor (SMO) belongs to the frizzled-class-G-protein coupled receptor and its activity can be classified in the canonical and the non-canonical pathway. The canonical pathway concludes in the transcriptional activation of the Hedgehog target genes through the GLI family of transcription factors whereas the non-canonical pathway leads to the recruitment and activation of trimeric G proteins. Recently, we have reported two novel roles of SMO with relevance to cell survival and drug resistance in DLBCL. In the first, SMO promotes a shift in the ubiquitination pattern for TNF receptor-associated factor 6 (TRAF6) from degradation inducing K48-ubiquitin branching towards K63-ubiquitination. TRAF6 is a K63-Ub E3 ubiquitin ligase for AKT and K63-ubiquitination of AKT increases membrane recruitment, activation and stability. Evidently, SMO promotes the formation of a scaffold-like protein complex including the de-ubiquitinating enzyme Ubiquitin specific peptidase 8 (USP8) and TRAF6. In its second novel role, SMO influences the vesicular sorting of IGF1R and other receptor tyrosine kinases. Loss of SMO leads to increased IGF1R lysosomal degradation and reduced IGF1R recycling, while overexpression increases IGF1R levels and AKT signaling. These observations indicate SMO receptor biology outside of the established canonical and non-canonical pathway functions. We have set out to use BioID to investigate the SMO interactome that contributes to these novel functionalities using a fusion protein of SMO and the reengineered E. coli biotinyl transferase BirA (Turbo) as bait. Preliminary BioID using SMO-Turbo returned a list of candidates which will require further data filtering approaches, but thus far shows a marked enrichment for proteins involved in vesicular trafficking.
Citation Format: Samantha Manuel, Nitin Agarwal, Daniel Bilbao, Francisco Vega, Ralf Landgraf. Exploring novel roles of the GPCR Smoothened in AKT signaling and drug resistance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2567.
Abstract 2582: Protein aggregation in sarcoma as a target for therapy
Authors: Sartini Sara , Le Hillary , Soragni Alice 
Published: April 2023

Abstract
Sarcomas are rare and heterogenous cancers arising from bone, soft and connective tissue. Developing effective therapies remains a challenge, since many types are understudied and poorly characterized at the molecular level. Here, we propose to investigate the role of protein misfolding in sarcoma, starting from the tumor suppressor p53. Wild type p53 is a tumor suppressor that regulates many cellular processes and prevents malignant transformation. Missense mutations promote p53 misfolding and convert the protein into a cancer-promoting oncogene. Mutant p53 is found in ~80% of all osteosarcoma cases, ~50% of leiomyosarcoma and angiosarcomas. It is also mutated at lower frequencies in other sarcoma subtypes. We investigated whether p53 mutations found in different types of sarcoma cause p53 misfolding and aggregation and can be targeted by ReACp53, a p53 aggregation-targeting peptide with anti-tumor activity in ovarian cancer and prostate cancer (Soragni et al, 2016, Zhang et al, 2019). We established tumor organoids from four sarcoma lines, which include osteosarcoma, fibrosarcoma, leiomyosarcoma and rhabdomyosarcoma. The cells carry either p53 mutations or WT p53. To characterize the p53 aggregation status and elucidate the response to ReACp53, we performed staining and quantification of folded/unfolded p53 pools in the organoids. Our results demonstrate the presence of overexpression and aggregation of p53 in the nuclei as well in the cytosol of different types of sarcoma lines. We found a marked susceptibility to ReACp53 in all lines, including cells bearing WT p53. We also explored whether similar p53 conformational defects are found in tumor tissue from sarcoma patients and if the activity of ReACp53 is confirmed in patient-derived tumor organoids. Overall, the discovery of structural alterations of p53 in sarcoma could provide an opportunity for the development of novel therapeutic strategies.
Citation Format: Sara Sartini, Hillary Le, Alice Soragni. Protein aggregation in sarcoma as a target for therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2582.
Abstract 2613: Chronic NNAL exposure, mimicking tobacco use in humans, deactivates LKB1 and promotes tumor progression in non-small cell lung cancer
Authors: Bian Tengfei , Jin Lingtao , Xing Chengguo 
Published: April 2023

Abstract
Background: LKB1 loss-of-function is one key oncogenic event in lung cancer. Clinical data suggest that LKB1 loss-of-function is associated with patients’ smoking status. We recently discovered that NNAL, a major metabolite of a tobacco specific carcinogen NNK, rapidly induces LKB1 phosphorylation and its loss-of-function via the β-AR/PKA signaling pathway in an isomer-dependent manner in human lung cancer cells. Such an acute exposure, however, only resulted in moderate enhancement in lung cancer cell migration and chemoresistance. Here we characterized the effects of chronic NNAL exposure, better mimicking human tobacco use, on LKB1 and tumor progression in NSCLC in vitro and in vivo.
Methods and Results: Human NSCLC cells with wild type LKB1 (H1299 and H1975) were cultured in the presence of 1 nM NNAL for 3, 6 and 12 months. LKB1 and its signaling pathways were characterized with phenotypes evaluated, including cell proliferation, migration, and chemoresistance. NNAL chronic exposure in lung cancer cells, mimicking its chronic exposure among smokers, resulted in more prominent LKB1 phosphorylation and the deactivation of its signaling, cell migration, and chemoresistance even in the absence of NNAL, indicating the long-lasting LKB1 loss-of-function. These observations were confirmed in a lung cancer xenograft model. More importantly, human lung cancer tissues revealed elevated LKB1 phosphorylation in comparison to the paired normal lung tissues.
Conclusion: These results suggest that LKB1 loss-of-function in human lung cancer could be extended to its phosphorylation, which may be mediated by NNAL from tobacco smoke in an isomer-dependent manner via the β-AR/PKA signaling pathway.
Citation Format: Tengfei Bian, Lingtao Jin, Chengguo Xing. Chronic NNAL exposure, mimicking tobacco use in humans, deactivates LKB1 and promotes tumor progression in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2613.
Abstract 2720: Enhancing oral delivery of plant-derived vesicles
Authors: Liu Yuan , Lankenau-Ahumada Adrian , Bayraktar Emine , Schwartz Paul , Chowdhury Mamur , Shi Sixiang , Sebastian Manu M. , Khant Htet , De Val Natalia , Jie Zuliang , Jennings Nicholas B. , Rodriguez-Aguayo Cristian , Swain Jody , Stur Elaine , Mangala Lingegowda S. , Wu Yutuan , Nagaraju Supriya , Erimas Brooke , Li Chun , Lopez-Berestein Gabriel , Braam Janet , Sood Anil K. 
Published: April 2023

Abstract
Plant-derived vesicles (PDVs) were attractive for therapeutic applications, including as potential nanocarriers. However, a concern with oral delivery of PDVs is whether they would remain intact in the gastrointestinal tract. We found that 82% of cabbage PDVs were destroyed under conditions mimicking the upper digestive tract. To overcome this limitation, we developed a delivery method whereby lyophilized Eudragit S-100-coated cabbage PDVs were packaged into a capsule (Cap-cPDVs). Lyophilization and suspension of PDVs in phosphate-buffered saline did not have an appreciable effect on PDV structure, number of PDVs, or therapeutic effect. However, packaging the lyophilized Eudragit S-100-coated PDVs into capsules allowed them to pass through the upper gastrointestinal tract for delivery into the colon better than did suspension of PDVs in phosphate-buffered saline. Cap-cPDVs demonstrated robust therapeutic effect in a dextran sulfate sodium-induced colitis mouse model. These findings could have broad implications for oral delivery of PDVs.
Citation Format: Yuan Liu, Adrian Lankenau-Ahumada, Emine Bayraktar, Paul Schwartz, Mamur Chowdhury, Sixiang Shi, Manu M. Sebastian, Htet Khant, Natalia De Val, Zuliang Jie, Nicholas B. Jennings, Cristian Rodriguez-Aguayo, Jody Swain, Elaine Stur, Lingegowda S. Mangala, Yutuan Wu, Supriya Nagaraju, Brooke Erimas, Chun Li, Gabriel Lopez-Berestein, Janet Braam, Anil K. Sood. Enhancing oral delivery of plant-derived vesicles [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2720.
Abstract 279: Lysosomal cyst(e)ine regulates melanoma de-differentiation and sensitivity to ferroptosis
Authors: Yu Deyang , Liang Jiaxin , Widlund Hans , Puigserver Pere 
Published: April 2023

Abstract
In response to anti-melanoma treatment, melanoma cells go through a process called de-differentiation that is characterized as loss of cellular features of melanocytes and acquisition of traits of the primitive neural crest stem cells which give rise to melanocytes during embryonic development. While de-differentiation enables melanoma to resist anti-melanoma drugs, it also exposes certain vulnerabilities of melanoma. A recent study showed that de-differentiated melanoma cells, while resistant to standard anti-melanoma therapies, display remarkable sensitivity to a group of chemicals that cause a special type of cell death called ferroptosis. However, the cellular programs and molecular machineries that drive melanoma de-differentiation and make them vulnerable to ferroptosis remain unknown. As the master regulator of melanocyte differentiation, MITF is often lost during de-differentiation and this loss has been correlated with sensitivity to ferroptosis. Through proteomic profiling of melanoma cells with different MITF status, we found that MITF-mediated lysosomal biogenesis is a key difference between differentiated melanoma cells and de-differentiated melanoma cells. Importantly, within the same melanoma linage, the heterogeneity in lysosomal abundance underlies differential sensitivity to ferroptosis. To probe lysosomal function that controls ferroptosis sensitivity, we deleted a series of lysosome-related proteins and found that the lysosomal cysteine transporters CTNS and MFSD12 exert opposing effects on lysosomal de-differentiation and ferroptosis sensitivity. Specifically, deletion of the pH-dependent cystine exporter CTNS drives melanoma de-differentiation and promotes ferroptosis sensitivity while ablation of cysteine importer MFSD12 further enhances resistance to ferroptosis. On the other hand, while increasing lysosomal cystine export by overexpression of CTNS has moderate effects on melanoma de-differentiation, increasing lysosomal important by overexpression of MFSD12 results in melanoma de-differentiation and increases sensitivity to ferroptosis. Our findings suggest that disruption of lysosomal cysteine homeostasis is a key mechanism to drive melanoma de-differentiation which is accompanied by increased sensitivity to ferroptosis.
Citation Format: Deyang Yu, Jiaxin Liang, Hans Widlund, Pere Puigserver. Lysosomal cyst(e)ine regulates melanoma de-differentiation and sensitivity to ferroptosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 279.
Abstract 2872: Site specific tumor microenvironments modulate the immune checkpoint inhibitors response in models of CT26 colon carcinoma
Authors: Zhao Fei , Thompson Caitlin , Barnes Nicole , Roques José , Coleman Terrence , Rowe Tyler , Kolb Joseph , Hall Chassidy , Fadden Patrick 
Published: April 2023

Abstract
Syngeneic models provide a robust platform to test new and cutting edge immune-oncology therapies. CT26 is a popular syngeneic colon carcinoma model that is known to respond to immune checkpoint inhibitors when engrafted into Balb/c mice as a subcutaneous flank tumor. The subcutaneous flank implant location provides a robust model, that is easy to set up, and allows for continuous measurement of the tumor growth by calipering. However, the location of tumor implant has been shown to impact tumor microenvironment and immune response for a number of cancer cell lines (Zhao 2017, Brand 2021, Oliver 2018). In addition to subcutaneous flank tumors, CT26 can be implanted to generate a lung metastasis or peritoneal metastasis model. We were interested to see if the lung metastasis or peritoneal metastasis CT26 model responded differently to immune checkpoint inhibition compared with the subcutaneous model and whether the different implant locations resulted in changes to the tumor infiltrating lymphocytes. CT26 cells were either injected intravenously to induce a lung metastasis model, intraperitoneally to produce a peritoneal metastasis model, or subcutaneously to generate a solid flank tumor model. Immune checkpoint inhibitor (ICI) therapy began on Day 5 following implant in each model, and tumor progression or survival provided the endpoint for analysis. In addition, tumor infiltrating leukocytes were assessed by flow cytometry to determine differences in the overall tumor microenvironment for each model. The response to both anti-PD-1 and anti-CTLA-4 monotherapies were strongest in the subcutaneous flank model, followed by moderate responses in the lung metastasis model, and no significant response to either ICI in the peritoneal metastasis model. This provides researchers with an opportunity to assess the ability of novel immune-oncology therapies to provoke an immunogenic response in models with identical antigenicity, but different tumor microenvironments based on the tumor location.
Citation Format: Fei Zhao, Caitlin Thompson, Nicole Barnes, José Roques, Terrence Coleman, Tyler Rowe, Joseph Kolb, Chassidy Hall, Patrick Fadden. Site specific tumor microenvironments modulate the immune checkpoint inhibitors response in models of CT26 colon carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2872.
Abstract 2882: Characteristic immunomodulatory role of myeloid galectin-9 in the brain tumor microenvironment
Authors: Lee Chanju , Yu Dahee , Park Eun Jung 
Published: April 2023

Abstract
Galectin-9 is a member of galectin family with carbohydrate-recognition domains that specifically bind to β-galactoside. Increasing reports have suggested that galectin-9 can bind and interact with immune-modulatory molecules including TIM-3 in several pathophysiological conditions, especially in T cells. However, to date, little is known about the galectin-9 in myeloid cells under tumor microenvironment. In this study, we explored the expressional characteristics and roles of galectin-9 in the brain tumor microenvironment using an intracranial mouse brain tumor model and in vitro system. We found that expression level of galectin-9 was markedly elevated in ipsilateral hemisphere compared to contralateral hemisphere of intracranial brain tumor model. FACS analyses showed that galectin-9 expression was higher in CD45highCD11bhigh macrophages and CD45midCD11bhigh microglia as well as lymphocytes from ipsilateral region than those from contralateral region. In addition, galectin-9 levels were increased after exposure to GL26 brain tumor cells in in vitro co-culture system using primary microglia. Interestingly, phagocytic activity, a key function of myeloid cells, was significantly reduced in galecin-9+ microglia compared to galecin-9- microglia when co-cultured with GL26-GFP cells. Furthermore, SIRPα levels were higher in galectin-9+ microglia than galecin-9- microglia. Similar results were observed in macrophages. Collectively, these findings suggest that galectin-9 may be an important modulatory molecule in myeloid cells under brain tumor microenvironment.
Citation Format: Chanju Lee, Dahee Yu, Eun Jung Park. Characteristic immunomodulatory role of myeloid galectin-9 in the brain tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2882.