bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 344: PKC-ι is a simultaneous modulator of the sonic hedgehog and the FAK/paxillin pathways in medulloblastoma
Authors: Lajmi Luke , Breedy Sloan , Todman Aaron , Ratnayake Wishrawana S. , Acevedo-Duncan Mildred
Published: April 2023

Medulloblastoma is the second most common childhood cancer, behind leukemia, and the most common pediatric brain tumor, comprising 20% of all childhood brain cancers. Our data demonstrated the effectiveness of atypical protein kinase C (aPKC) inhibitors 2-acetyl-1,3-cyclopentanedione (ACPD), 3,4-diaminonapthalene-2,7-disulfonic acid (DNDA), 5-amino-1-2,3-dihydroxy-4-(methylcyclopentyl)-1H-imidazole-4-carboxamide (ICA-1S), and 8-hydroxy-1,3,6-naphthalenetrisulfonic acid (ζ-Stat) to significantly decrease cell proliferation with increasing inhibitor concentration. The continuation of this study involves examining the oncogenic role of PKC-ι as a key molecule that acts as a central modulator of the activities of the sonic hedgehog (SHH) pathway and the focal adhesion kinase (FAK)/paxillin pathway. Our data will demonstrate the toxic effects of ICA-1S, more so than the other three inhibitors, through WST-1 assays as well as Western blot data that shows the effectiveness of ICA-1S to selectively downregulate the phosphorylation of PKC-ι, by 42% in D283 cells and 20% in Daoy cells, which resonantly impacts the various signaling proteins that govern the SHH and FAK/paxillin pathway across D283, D341, and Daoy medulloblastoma cell lines. Phosphorylation of FAK was reduced in D283 by 25%, 30% in D341 cells, and 15% in Daoy cells upon treatment with ICA-1S. This indicates that even though these cell lines have different mutations, they appear to proliferate by similar signaling pathways. Immunoprecipitation studies showed that simultaneous downregulation of PKC-ι and PKC-ζ in the D341 cell line is the result of the association of PKC-ι and PKC-ζ, indicating they may directly regulate each other’s catalytic activities. Furthermore, ICA-1S was shown to reduce the association of PKC-ι and PKC-ζ by over 75% in D341 cells. Future studies will be conducted to determine the effect of aPKC inhibitors on additional downstream proteins in the FAK/paxillin pathway and cell cycle regulation by Western blotting. We will also be investigating the effect of the inhibitors on the migratory and invasive capacity of medulloblastoma by implementing wound-healing assays and Boyden chamber assays.

Citation Format: Luke Lajmi, Sloan Breedy, Aaron Todman, Wishrawana S. Ratnayake, Mildred Acevedo-Duncan. PKC-ι is a simultaneous modulator of the sonic hedgehog and the FAK/paxillin pathways in medulloblastoma [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 344.
Abstract 346: Downregulating KIF4A inhibited growth of uterine leiomyosarcoma by regulating cell cycle
Authors: Nakagawa Satoshi , Mizuta-Odani Chihiro , Kobayashi Mariya , Kakuda Mamoru , Hiramatsu Kosuke , Kimura Toshihiro , Kobayashi Eiji , Ueda Yutaka , Kimura Tadashi
Published: April 2023

[Objective] Uterine leiomyosarcoma (LMS) has a poor prognosis, and no effective treatment other than surgery has been established. Kinesin family member4A (KIF4A) is a member of the kinesin4 subfamily and plays an important role in cell division. KIF4A has been revealed to play significant roles in some cancers with tumor proliferation. The aim of this study is to clarify the relationship between KIF4A and tumor growth of LMS.

[Methods] One normal human uterine smooth muscle cell line and three LMS cell lines(SK-UT1, SK-LMS and SKN) were used to identify tumor-specific elevated proteins with iTRAQ(isobaric tags for relative and absolute quantitation). SiRNA and ShRNA were used to downregulate KIF4A in vitro and in in vivo. Cell variability was evaluated by MST-8 assay. Cell cycle analysis by fluorescence-activated cell sorting (FACS) and western blotting (WB) were performed to analyze the signal pathway. Immunostaining of 26 clinical specimen from patients operated at our university hospital since 1999 to 2021.

[Results] A total of 2084 proteins were identified by iTRAQ. KIF4A was identified as a protein with more than twice the expression level of normal smooth muscle cells. WB confirmed high expression of KIF4A in all three LMS cell lines. And in immunostaining of clinical specimens, KIF4A was positive in 20/26(76.9%) cases. A trend toward worse prognosis was suggested in the group with high KIF4A expression (no significant difference due to the small number of cases). Downregulation with SiRNA significantly suppressed the growth of LMS cell lines in vitro (-37.2±4.73% in SK-LMS, -87.7±4.02% in SKN and -28.1±3.00% in SK-UT1, p<0.05). By FACS, G2/M phase cells was significantly increased in KIF4A suppressed cell lines and by WB, p-cdc2 was upregulated in KIF4A suppressed cells. In vivo, downregulation of KIF4A was found to significantly suppress tumor size in genograft mice. (1186±118mm³ vs 461±84mm³ in SK-LMS and 1704±441mm³ vs 514±230mm³ in SK-UT1, p<0.05, f).

[Conclusion] Downregulating KIF4A inhibited LMS tumor growth and it can be a novel therapeutics target to treat LMS.

Citation Format: Satoshi Nakagawa, Chihiro Mizuta-Odani, Mariya Kobayashi, Mamoru Kakuda, Kosuke Hiramatsu, Toshihiro Kimura, Eiji Kobayashi, Yutaka Ueda, Tadashi Kimura. Downregulating KIF4A inhibited growth of uterine leiomyosarcoma by regulating cell cycle [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 346.
Abstract 349: LncRNA Myca controls hepatocellular carcinoma growth and progression
Authors: Chen Hsin-Yi , Weng Yu-Yun , Yuan Wei-Chien , Lee Yu-Ru
Published: April 2023

Long non-coding RNAs (lncRNAs) have been shown to play critical roles in the tumorigenesis and progression of cancer. From a clinic and genomic perspective of evidence, the MYC signaling plays a crucial role in triggering hepatocellular carcinoma (HCC) and therefore represents an attractive target for HCC intervention. However, no specific drug is currently available to target MYC directly due to the lack of an enzymatic pocket. Therefore, deciphering the critical lncRNAs that functionally crosstalk with MYC protein levels may offer novel therapeutic strategies in HCC. Here we identify the involvement of a lncRNA Myca (Myc Activator) in HCC through controlling MYC protein levels. Overexpression of Myca triggers tumor growth in a MYC-dependent manner, while inactivation of Myca by antisense oligonucleotides (ASOs) profoundly reduces MYC protein levels, thus suppressing tumor progression in vitro. Mechanistically, we show that Myca attenuates MYC ubiquitination and leads to MYC accumulation. Clinically, overexpression of Myca is positively correlated with Hi-MYC transcriptional profile and HCC progression from the TCGA dataset. Together, our findings not only identify Myca in controlling MYC protein levels but also provide Myca as a new therapeutic lncRNA target for the HCC intervention.

Citation Format: Hsin-Yi Chen, Yu-Yun Weng, Wei-Chien Yuan, Yu-Ru Lee. LncRNA Myca controls hepatocellular carcinoma growth and progression [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 349.
Abstract 3512: Organ-specific effects of Smad4 in metastatic pancreatic cancer
Authors: Tsanov Kaloyan M. , Barriga Francisco M. , Ho Yu-Jui , Alonso-Curbelo Direna , Livshits Geulah , Koche Richard , Baslan Timour , Wuest Alexandra N. , Simon Janelle , Tian Sha , Luan Wei , Lowe Scott W.
Published: April 2023

Primary and metastatic tumors typically share the same driver gene mutations, but it is unclear if these mutations are functionally relevant across different anatomical sites. Among such mutations, inactivation of the tumor suppressor gene SMAD4 is a hallmark of pancreatic and other gastrointestinal cancers and has been associated with metastatic disease. Here, we develop a mouse model of pancreatic ductal adenocarcinoma that enables Smad4 depletion in the pre-malignant Kras-mutant pancreas and subsequent Smad4 reactivation in late-stage metastatic tumors. Whereas early Smad4 deficiency facilitated tumor formation, later Smad4 restoration had unexpected organ-specific outcomes: no effect on primary tumor growth, suppression of liver metastases, and promotion of lung metastases. Integrative multiomic analysis revealed a near-universal genomic deletion of the Cdkn2a/b locus and organ-specific changes in the tumor cells’ epigenomic state. In particular, the liver and lung differentially favored KLF vs. RUNX dominated chromatin programs, which were confirmed to underpin the divergent effects of Smad4 restoration in functional studies. Our results show how organ-dependent epigenomic changes lead to altered driver gene function in metastatic disease. This organ-specific gene-chromatin interplay may be a generalizable principle in cancer biology and invites a revised paradigm for precision oncology that considers anatomical site in the interpretation of tumor genetics.

Citation Format: Kaloyan M. Tsanov, Francisco M. Barriga, Yu-Jui Ho, Direna Alonso-Curbelo, Geulah Livshits, Richard Koche, Timour Baslan, Alexandra N. Wuest, Janelle Simon, Sha Tian, Wei Luan, Scott W. Lowe. Organ-specific effects of Smad4 in metastatic pancreatic 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 3512.
Abstract 3867: Chromatin modification driving sub-clonal resistance to KRAS G12C combination therapies in KRAS mutant non-small cell lung cancer
Authors: Li Chendi , Qin Qian , Syed Mohammed Usman , Nimbalkar Anahita , Karakyriakou Barbara , Clark Sarah E. , Saiki Anne Y. , Hughes Paul E. , Ott Chris , Pinello Luca , Hata Aaron N.
Published: April 2023

The FDA approval of the KRAS G12C inhibitor (G12Ci) sotorasib and the advancement of similar drugs into clinical trials marks a major milestone in treating KRAS G12C non-small cell lung cancer (NSCLC). However, not all patients respond (sotorasib - ORR = 37.1%, adagrasib - 43%, JDQ443 - 35%), motivating preclinical and clinical investigation into mechanisms of intrinsic and acquired resistance. For instance, clinical studies have reported on-target KRAS mutations and preclinical studies have demonstrated mitogen-activated protein kinase (MAPK) feedback reactivation including EGFP, SHP2, and WT RAS signaling. In response to targeted therapies, sub-populations of cells can enter quiescence or specific epigenetic-driven states that confer drug tolerance. However, epigenetic states defining drug-tolerant persister populations and contributing to adaptive resistance to KRAS G12Ci have not been reported. Using a lineage tracing barcoded system, we identify distinct and reversible subpopulations defined by specific chromatin and transcriptional states in KRAS NSCLC cell lines that contribute to KRAS G12Ci resistance in vitro, even prior to drug treatment. We observed that specific states, including activation of histone demethylation and SWI/SNF complex, may contribute to MAPK reactivation-driven resistance. These results suggest potential epigenetic vulnerabilities that can be exploited to improve the response to KRAS G12Ci. Moreover, we observed distinct persister subpopulations with resistance to KRAS G12Ci combination co-targeting orthogonal pathways (SHP2, CDK4/6, PI3K, and MCL-1), raising the possibility that distinct epigenetic-transcriptional states contribute to differential drug response and clonal evolution of persisters. Collectively, these results suggest that more complete tumor regression may be achieved by orthogonal strategies that target different resistant populations within the same tumor.

Citation Format: Chendi Li, Qian Qin, Mohammed Usman Syed, Anahita Nimbalkar, Barbara Karakyriakou, Sarah E. Clark, Anne Y. Saiki, Paul E. Hughes, Chris Ott, Luca Pinello, Aaron N. Hata. Chromatin modification driving sub-clonal resistance to KRAS G12C combination therapies in KRAS mutant 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 3867.
Abstract 388: Acquired sensitivity to ONC201/TIC10 in ER+ breast cancer progressing on the mTOR inhibitor everolimus
Authors: Farmaki Eleni , Emond Rena , Nath Aritro , Grolmusz Vince K. , Cosgrove Patrick A. , Bild Andrea H.
Published: April 2023

The mTOR inhibitor everolimus has become an important component for clinical management of metastatic ER+ breast cancer. Despite the improved patient outcomes, progression and resistance will still develop. Therefore, it is important to research strategies that can increase response durability and effectiveness of this therapy. Here we developed everolimus resistance models and performed pre-clinical evaluation of the small molecule ONC201/TIC10, a drug candidate showing promising results in clinical trials with advanced cancer patients. ONC201/TIC10 targets mitochondrial function and metabolism and activates stress response, however its efficacy has not been studied in everolimus resistant breast cancer. Transcriptomic analysis in everolimus resistant tumors identified sustained mitochondrial oxidative phosphorylation activity suggesting increased sensitivity to ONC201/TIC10. Our results show that combination treatment of ONC201/TIC10 with everolimus inhibits the growth of sensitive as well as everolimus resistant cells in 2D and 3D in vitro studies. The increased response to combination therapy was confirmed in primary cells from patients progressing on endocrine and mTOR therapy, supporting clinical relevance. We further show that the mechanism of ONC201/TIC10 anti-proliferative activity involves disruption of mitochondrial function and increased levels of the pro-apoptotic transcription factors CHOP and ATF4. Importantly, several resistant cell lines demonstrated increased dependency on mitochondrial respiration compared to sensitive cells which contributes to a vulnerability to ONC201/TIC10. In conclusion, we used pre-clinical models to characterize acquired sensitivity to mTOR inhibition. Combining patient derived data with resistant cell line driven experiments, our study provides validated findings in various settings strengthening the potential for clinical applications of ONC201/TIC10. We propose that ONC201/TIC10 and modulation of mitochondrial function is effective in drug resistant cancer cells and can enhance the anti-tumor activity of standard of care therapy in progressing patients.

Citation Format: Eleni Farmaki, Rena Emond, Aritro Nath, Vince K. Grolmusz, Patrick A. Cosgrove, Andrea H. Bild. Acquired sensitivity to ONC201/TIC10 in ER+ breast cancer progressing on the mTOR inhibitor everolimus [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 388.
Abstract 3888: N-glycosylation-defective IL6 activates the SRC-YAP-SOX2 signaling to potentiate metastasis and TKI resistance in NSCLC
Authors: Hung Chun Hua , Wu Shang-Yin Wu , Yeh Hsuan-Heng Yeh , Lin Chien-Chung , Su Wu-Chou
Published: April 2023

The IL6-GP130-STAT3 signaling pathway facilitates lung cancer progression and resistance to tyrosine kinase inhibitors. Although glycosylation alters the stability of GP130 receptor, its effect on the ligand IL6 remains unclear. In this study, we found that N-glycosylated IL6 primarily triggers JAK-STAT3 signaling and prolongs STAT3 phosphorylation, whereas N-glycosylation-defective IL6 (DeNG-IL6) induces shortened STAT3 activation and changes the downstream signaling preference for the SRC-YAP-SOX2 axis. This signaling shift promoted epithelial plasticity in vitro and metastasis in vivo, which were suppressed by specific inhibitors and shRNAs targeting SRC, YAP, and SOX2. EGFR TKI-resistant lung cancer cells secrete high levels of DeNG-IL6 via reduced N-glycosyltransferase gene expression and showed SRC-YAP activation. DeNG-IL6 contributes to drug resistance, as confirmed by in silico analysis of cellular and clinical transcriptomes, and signal expression in patient specimens. In summary, cell behaviors can be altered by modifying the N-glycosylation of IL6, and the glycosylation status of circulating IL6 might be a promising biomarker for monitoring the dynamics of lung cancer evolution.

Citation Format: Chun Hua Hung, Shang-Yin Wu Wu, Hsuan-Heng Yeh Yeh, Chien-Chung Lin, Wu-Chou Su. N-glycosylation-defective IL6 activates the SRC-YAP-SOX2 signaling to potentiate metastasis and TKI resistance in NSCLC. [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 3888.
Abstract 3889: The role of CXCL1 in crosstalk between breast cancer cells with ESR1 mutations and lymphatic endothelial cells
Authors: Galke Daniel , Jin Kideok
Published: April 2023

Various endocrine therapies have been developed such as selective estrogen receptor modulation (SERM), selective estrogen receptor downregulator (SERD) and ligand deprivation using aromatase inhibitors (AI). Tamoxifen, fulvestrant, and AIs are the most used adjuvant treatment for pre- and postmenopausal women with early-stage estrogen receptor positive breast cancer. Tamoxifen and AIs have widespread anti-neoplastic and chemopreventive effects, yet despite this, many breast cancers that are initially receptive ultimately become resistant and recur. There are many known underlying molecular processes that result in resistance, but no overarching pattern has yet been identified. Our previous research demonstrates that the crucial molecular and cellular players secreted from a crosstalk between stromal cells and breast cancer cells can control the development of the tumor and the metastatic process in breast cancer. Breast cancer metastasis involves lymphatic dissemination in addition to hematogenous spreading. Although stromal lymphatic vessels (LVs) serve as initial metastatic routes, roles of organ-residing LVs are underinvestigated. In this report, we demonstrated that crosstalk between ESR1 mutant cells (Y537S and D538G), and lymphatic endothelial cells (LECs), a component of lymphatic vessels, promoted ESR mutant cell growth and migration. To identify a critical secreted factor in this crosstalk, we performed the cytokine array in secretome of crosstalk between ESR1 mutant cells and LECs, and found that CXCL1 is highly secreted from LECs. Knockdown of CXCR2 in ESR1 mutant cells attenuated ESR1 cell migration in the conditioned media of LECs crosstalked with ESR1 mutant cells. To investigate a potential impact given the current clinical landscape of endocrine resistant breast cancer with ESR1 mutations, we performed a cell viability assay and migration assay using reparixin, CXCR1/2 inhibitor in co-culture with ESR1 mutant cells and LECs. Significantly, reparixin decreased ESR1 mutant cell viability and migration compared to vehicle. We found that the combination treatment of reparixin and Palbociclib and ribociclib significantly inhibited ESR1 mutant cell growth and migration compared the single treatment. These findings implicate CXCL1-CXCR1/2 signaling as a critical event in the growth of breast cancer with ESR1 mutation and metastasis. Therefore, we have provided evidence that supports the hypothesis that functional inhibition of the CXCL1 and CDK4/6 signaling pathway has the potential to circumvent endocrine resistant tumor growth and metastasis.

Citation Format: Daniel Galke, Kideok Jin. The role of CXCL1 in crosstalk between breast cancer cells with ESR1 mutations and lymphatic endothelial cells. [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 3889.
Abstract 3898: Synergistic combination of the iron chelator deferasirox with cisplatin and doxorubicin chemotherapies against nonsmall cell lung carcinoma
Authors: Alvelo Natalia I. Ortiz , Torres Grace , Estrada Sthephanie , Pérez Daraishka , Ferrer Yancy , Delgado Yamixa
Published: April 2023

Lung cancer is the second most common and leading cause of cancer-related deaths worldwide with 85% of these cases being non-small cell lung carcinoma (NSCLC). Unfortunately, despite improvements in NSCLC outcomes, many of these tumors develop resistance against principal chemotherapies (e.g., cisplatin (CisPt) and doxorubicin (Doxo)), inducing abysmally low survival prognosis. Deferasirox (Def), an FDA-approved iron chelation therapy, is a drug under study for cancer therapy due to iron plays a key role in cell growth and energy production. Herein, we determined the effect of Def alone and in combination with CisPt and Doxo on NSCLC A549 cells. Viability assays results showed that Def has synergistic cytotoxic effects in combination with CisPt and Doxo at low uM concentrations after 24 h of incubation using the Chou Talalay method. In addition, qPCR gene expression studies showed that Def induced a significant downregulation of EGFR, VEGF, MMP9, MMP2, CHD4 genes related to resistance and metastatic process. In addition, we determined the overexpression of NDRG1 gene mediated by Def confirming the chelation of iron and disruption of iron metabolism. Overall, this work will set the basis for adjuvant therapies against chemoresistance and metastasis processes using an iron chelator.

Citation Format: Natalia I. Ortiz Alvelo, Grace Torres, Sthephanie Estrada, Daraishka Pérez, Yancy Ferrer, Yamixa Delgado. Synergistic combination of the iron chelator deferasirox with cisplatin and doxorubicin chemotherapies against nonsmall cell lung 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 3898.
Abstract 3899: CHST11 mediated SEMA7A expression contributes to TKI resistant in lung cancer
Authors: Li Chien-Hsiu Chris , Chan Ming-Hsien , Chang Yu-Chan , Hsiao Michael
Published: April 2023

An extracellular matrix has been demonstrated to be one of the substances that promote tumor cells to become driven by external stimuli to change their biology or malignancy. The main component of extracellular matrix, chondroitin sulfate, has been confirmed as a factor in the progression of tumors. Nevertheless, a great deal remains unknown regarding the precise mechanisms that drive Chondroitin sulfate production and how these details are communicated to cancer cells. According to our previous studies, the main enzyme responsible for Chondroitin sulfate synthesis, carbohydrate sulfotransferase 11 (CHST11), plays a crucial role in lung cancer cell metastatic spread. Nevertheless, the relationship between CHST11 expression and regulatory mechanism in response to lung cancer treatment is unclear, especially for tyrosine kinase inhibitor (TKI). A clinical correlation analysis was used in this study to identify molecules which were highly correlated with the overexpression of CHST11 and identified GPI-anchored protein semaphorin 7A (SEMA7A) as a candidate. According to the TCGA database, expression of upregulated SEMA7A is correlated with poor prognosis and cellular malignancy in lung cancer patients. A SEMA7A-based gene correlation analysis finds that the affected gene ontology is highly similar to CHST11, particularly integrin signaling, which may contribute to Gefitinib resistance. A GSEA analysis reveals a significant effect of CHST11 overexpression on mitotic spindle processes, some of which molecules are clinically relevant to CHST11 or SEMA7A. These molecules have been reported to be involved in TKI resistance events such as MARCKS, MYO1E, and PDLIM5. Based on the TCGA database, these molecules and integrins also display significant clinical relevance in lung cancer patients. A CHST11-mediated increase in SEMA7A in the mitotic spindle process of cells may serve as one of the possible means of reducing TKI-induced mitotic cell death, and thereby promoting the ability of lung cancer cells to become resistant to TKIs. Hence, targeting the CHST11-SEMA7A axis may offer a therapeutic strategy option that could be used to target drug resistance in lung cancers. [Keywords: CHST11, SEMA7A, Integrin, Drug resistant, Lung cancer] (Reference: https://pubmed.ncbi.nlm.nih.gov/35985204/; https://pubmed.ncbi.nlm.nih.gov/36181245/)

Citation Format: Chien-Hsiu Chris Li, Ming-Hsien Chan, Yu-Chan Chang, Michael Hsiao. CHST11 mediated SEMA7A expression contributes to TKI resistant in 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 3899.
Abstract 3903: Role of the miR-301a/Fra-2/GLIPR1 axis in lung cancer cisplatin resistance
Authors: Lovat Francesca , Vinciguerra Gian Luca Rampioni , Capece Marina , Distefano Rosario , Nigita Giovanni , Vecchione Andrea , Croce Carlo M.
Published: April 2023

Lung cancer is the leading cause of cancer-related mortality in both men and women worldwide. Despite the administration of platinum compounds represents the main treatment, the majority of tumors remains intrinsically resistant. In lung cancer, miR-301a exerts a main role by modulating the activity of transcription factors like NF-kB and Stat3. Here, we dissected the contribution of miR-301a in the regulation of its new identified target Fos-related antigen-2 (Fra-2), a transcription factor that belongs to the Fos/AP-1 family. By microarray analysis, we discovered that Fra-2 overexpression increased glioma pathogenesis-related protein 1 (GLIPR1) levels, a mediator of cisplatin resistance in lung cancer. Its mechanism of regulation is still unclear. We demonstrated that GLIPR1 is also a miR-301a target. In vitro, modulation of miR-301a reduced Fra-2 and GLIPR1 levels; conversely, Fra-2 overexpression significantly increased GLIPR1 and miR-301a. Then, by chromatin immunoprecipitation assay, we confirmed that Fra-2 interacts with the promoter regions of both GLIPR1 and MIR301A genes, supporting that miR-301a/Fra-2/GLIPR1 axis could be autoregulated by feedback loop in which Fra-2 promotes the transcription of MIR301A gene. Investigating the effect of this new axis on the response to cisplatin, we observed that both Fra-2/GLIPR1 overexpression and miR-301a silencing induced cisplatin resistance in lung cancer cells. By contrast, silencing of GLIPR1 and combined administration of low doses of Fos/AP-1 inhibitor restored the cisplatin sensitivity in Fra-2 overexpressing cells. In the lung adenocarcinoma samples from the TCGA dataset, miR-301a overexpression inversely correlated with Fra-2 and GLIPR1 expression. Consistently, the overexpression of miR-301a identified a fraction of tumors with low expression of Fra-2 and GLIPR1 in an internal cohort of lung cancer samples. Altogether, our findings identify the miR-301a/Fra-2/GLIPR1 axis that contributes to cisplatin resistance in lung cancer cells and could serve as biomarker to stratify patients who may benefit from cisplatin administration, alone or in combination with Fos/AP-1 inhibitors.

Citation Format: Francesca Lovat, Gian Luca Rampioni Vinciguerra, Marina Capece, Rosario Distefano, Giovanni Nigita, Andrea Vecchione, Carlo M. Croce. Role of the miR-301a/Fra-2/GLIPR1 axis in lung cancer cisplatin 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 3903.
Abstract 3905: Epigenetic primming reveals the central role of epigenetic state in the anti-tumor activity of vitamin D in pancreatic ductal adenocarcinoma
Authors: He Bo , Stoffel Lauren , He Clifford Jiajun , Li Albert Mao , Jiang Haowen , Cho Kumsun , Flowers Brittany M. , Ferreira Sofia , Attardi Laura D. , Ye Jiangbin
Published: April 2023

Pancreatic ductal adenocarcinoma (PDAC) has a dismal prognosis, which is largely due to the intrinsic resistance to a broad spectrum of therapies. Vitamin D (VD) has been extensively studied as a potential preventive or therapeutic agent for PDAC treatment. Although accumulating evidence supports that VD has beneficial effects on PDAC treatment, controversies still exist. The major reason for this inconsistency is that VD resistance widely occurs in PDAC. However, the underlying mechanism by which the VD resistance occurs remains largely unknown. Here, we report that epigenetic primming by glyceryl triacetate (GTA) and 5-azacytidine (5-Aza) overcomes VD resistance in PDAC. Mechanistically, increasing global H3K27 acetylation with GTA and reducing global DNA methylation with 5-Aza not only elevate the VDR expression but also reprogram the VD responsive genes. Consistently, combination treatment with VD analogue calcipotriol (CPT) and epigenetic primming overcomes the VD resistance to inhibit cell proliferation and migration through activating genes involved in negative regulation of cell proliferation and cell migration, while CPT treatment alone has the opposite effects. Intriguingly, further studies in mouse PDAC cells suggest that VDR expression does not always corelate with VD responsiveness. Using the gain and loss of function experiments, we demonstrate that VDR is necessary but not sufficient to trigger the anti-tumor activity of VD, highlighting that epigenetic state is the key determinant of VD responsiveness in PDAC. Collectively, these data reveal a previously undefined mechanism for VD resistance in PDAC and has the potential to guide VD clinical trials in the future.

Citation Format: Bo He, Lauren Stoffel, Clifford Jiajun He, Albert Mao Li, Haowen Jiang, Kumsun Cho, Brittany M. Flowers, Sofia Ferreira, Laura D. Attardi, Jiangbin Ye. Epigenetic primming reveals the central role of epigenetic state in the anti-tumor activity of vitamin D in pancreatic ductal adenocarcinoma. [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 3905.
Abstract 3936: ALK fusion oncogene driven SERPINB4 expression enhances tumor survival in NSCLC
Authors: Chuang Tzu-Po , Lai Wei-Yun , Gabre Jonatan L. , Lind Dan E. , Umapathy Ganesh , Bokhari Abdulmalik A. , Bergman Bengt , Kristenson Linnea , Thorén Fredrik B. , Le Anh , Doebele Robert , Eynden Jimmy V. , Palmer Ruth H. , Hallberg Bengt
Published: April 2023

Anaplastic lymphoma kinase (ALK) fusion variants in non-small-cell-lung cancer (NSCLC) consist of numerous dimerising fusion partners, with the most common being EML4. Clinical data suggests that the degree of treatment benefit in response to ALK tyrosine kinase inhibitors (TKIs) differs among the variant present in the patient tumor. Therefore, a better understanding the oncogenic signaling networks driven by different ALK-fusion variants is important. Here, we developed highly controlled doxycycline-inducible cell models bearing four different ALK fusion proteins, namely EML4-ALK-V1, EML4-ALK-V3, KIF5B-ALK, and TFG-ALK, in the context of non-tumorigenic NL20 human bronchial epithelial cells. These were complimented by patient-derived NSCLC cell lines harboring either EML4-ALK-V1 or EML4-ALK-V3 fusions. RNA-seq and phosphoproteomics analysis were employed to identify dysregulated genes and hyper/hypo-phosphorylated proteins associated with ALK fusion expression. Among ALK fusion induced responses, we noted a robust inflammatory signature that included up-regulation of the Serpin B4 serine protease inhibitor in both NL20-inducible cell models and ALK-positive NSCLC patient-derived cell lines. We show that STAT3 is a major transcriptional regulator of SERPINB4 downstream of ALK fusions, along with NF-kB and AP1. The upregulation of SERPINB4 promotes survival of ALK fusion expressing cells and inhibits natural killer (NK) cell-mediated cytotoxicity. In conclusion, our study reveals a novel ALK downstream survival axis that regulates Serpin B4 expression and identifies a molecular target that has potential for therapeutic impact targeting the immune response together with ALK TKIs in NSCLC.

Citation Format: Tzu-Po Chuang, Wei-Yun Lai, Jonatan L. Gabre, Dan E. Lind, Ganesh Umapathy, Abdulmalik A. Bokhari, Bengt Bergman, Linnea Kristenson, Fredrik B. Thorén, Anh Le, Robert Doebele, Jimmy V. Eynden, Ruth H. Palmer, Bengt Hallberg. ALK fusion oncogene driven SERPINB4 expression enhances tumor survival in NSCLC. [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 3936.
Abstract 3994: Differential regulation of hormone receptors and Ki67 in luminal breast carcinomas and tumor-adjacent mammary glands by mifepristone treatment
Authors: Saldain Leo , Vanzulli Silvia , Vazquez Paula Martinez , Burruchaga Javier , Spengler Eunice , Lanari Claudia , Rojas Paola
Published: April 2023

Since antiprogestins inhibit the growth of preclinical mammary tumor models expressing higher levels of progesterone receptor (PR) isoform A (PRA) than isoform B (PRB), we designed a window-of-opportunity trial (MIPRA; NCT02651844) to study the benefits of mifepristone (MFP) in luminal breast carcinomas from postmenopausal patients selected by their PR+ expression (>50%) and their PR isoform ratio (PRA/PRB>1.5; PRA-H). A decrease in the cell proliferation marker Ki67 was registered after treatment in 14 out of the 20 tumors evaluated. This inhibition was associated with a decrease in PR, estrogen receptor (ER) and pSer118ER evaluated by immunohistochemistry while no changes in pSer167ER expression were observed (PMID: 36269797). In selected tissues from these samples we observed that the staining intensity in trapped glands within the MFP-treated tumors did not follow a similar trend as that of tumor cells. Thus, the aim of this study was to evaluate the expression of hormone receptors and Ki67 in non-neoplastic human mammary glands (nnMG) adjacent to the PRA-H tumor tissue of postmenopausal patients, treated (n=8) or not (n=9) with MFP. No differences in PR, ER and pSer118ER expression were observed between MFP-treated nnMG and those from untreated patients. Notably, contrarily to what occurred in tumors, there was a significant decrease of pSer167ER expression (p=0.008) in the MFP-treated nnMG compared to the untreated glands, suggesting a selective modulation in the AKT-mediated activation of ER. Regarding the proliferative state of the nnMGs, there was a slight but significant increase (p=0.02) in Ki67 expression in MFP-treated vs. non-treated nnMG. Data regarding the effect of antiprogestins in normal mammary glands is limited and points, in premenopausal women, to a decrease in the Ki67 index after treatment. The basal quiescent status of nnMG in postmenopausal women may explain this slight stimulatory effect. In conclusion, our results show that MFP exerts a specific regulatory effect in PRA-H tumors that is not observed in the nnMG, probably due to the fact that nnMG have presumably equimolar levels of PRA and PRB. In addition, to counteract the possible stimulatory effect that MFP may induce on nnMG, the combination of MFP and tamoxifen treatment is suggested.

Citation Format: Leo Saldain, Silvia Vanzulli, Paula Martinez Vazquez, Javier Burruchaga, Eunice Spengler, Claudia Lanari, Paola Rojas. Differential regulation of hormone receptors and Ki67 in luminal breast carcinomas and tumor-adjacent mammary glands by mifepristone treatment. [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 3994.
Abstract 4071: Identification of myeloma-specific T cell receptors by functional single cell interaction analyses
Authors: Wagner Tim R. , Boğa Eren , Schmidt Patrick , Osen Wolfram , Platten Michael , Goldschmidt Hartmut , Raab Marc S. , Friedrich Mirco J. , Eichmüller Stefan B.
Published: April 2023

Innovative immunotherapy approaches such as adoptive transfer of chimeric antigen receptor (CAR) T cells or tumor infiltrating lymphocytes (TILs) have shown great success in the treatment of solid tumors and hematological malignancies. Although treatment of multiple myeloma with CAR T cells can induce deep responses, relapses frequently occur due to antigen escape and limited CAR T cell persistence. TCR-engineered T cells may mediate sustained antitumor effects upon recognition of intracellular targets, thereby significantly increasing the range of relevant target antigens. In our project, we propose to identify T cell receptors (TCRs) specifically targeting autologous myeloma cells. Tumor-reactive T cells were identified using the Berkeley Lights Lightning platform, allowing simultaneous functional analysis of up to 1500 individual T cell/target cell interactions per run. Reactive T cells have been identified upon detection of secreted chemokines (IFNγ, TNFα, IL-2) and by measurement of CD137 surface expression. Tumor-reactive T cells showing various cytokine secretion patterns and CD137 expression profiles could be detected in each myeloma patient (7 to 26 of approx.1200 cells tested per patient). Individual tumor reactive T cells have been isolated for TCR sequencing. Recovered TCR genes will be cloned and overexpressed in autologous T cells for functional validation and analysis of tumor derived neoepitope specificity. In summary, we present a pipeline allowing identification of myeloma-recognizing T cells and recovery of bona fide tumor-reactive TCRs eligible for patient-individualized T cell therapy.

Citation Format: Tim R. Wagner, Eren Boğa, Patrick Schmidt, Wolfram Osen, Michael Platten, Hartmut Goldschmidt, Marc S. Raab, Mirco J. Friedrich, Stefan B. Eichmüller. Identification of myeloma-specific T cell receptors by functional single cell interaction analyses. [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 4071.
Abstract 4089: PanCAR-specific antibody-cytokine fusion proteins
Authors: Keane John T. , Posey Avery D.
Published: April 2023

CAR-T cells are a potent immunotherapy that redirect T cell specificity towards cell surface tumor-associated antigens. While this therapy has been effective in hematological malignancies, therapies against solid tumors not achieved the same efficacy due to a multitude of additional challenges, including a lack of tumor specific antigens and an immunosuppressive tumor microenvironment. One strategy to increase T cell potency is to use fourth generation CAR-T cells that also secrete an immunostimulatory factor, such as a cytokine. We have created CAR-T cells targeting Tn-MUC1 utilizing a scFv from the 5E5 antibody that also secrete interleukin-12 (IL-12), interleukin-18 (IL-18), and interleukin-23 (IL-23). These cytokine-secreting CAR-T cells have shown an increase in efficacy against multiple epithelial based tumors in impedance-based killing assays. In a xenograft model of human breast cancer, CAR-T cells secreting IL-12 and IL-23 were able to significantly delay tumor growth compared to non-transduced T cells, CAR-T cells alone, and CAR-T cells secreting IL-18. While these cells showed good efficacy in vivo, clinical translation is difficult due to concerns over cytokine storm due to the constitutive secretion of cytokines. To overcome this, we created a fusion protein utilizing an antibody specific to an invariant region found in majority of CAR molecules, including those utilized commercially available therapies and under pre-clinical development, fused to the three immunostimulatory cytokines. In impedance-based killing assays, antibody-cytokine fusions stimulated non-cytokine secreting CAR-T cells to completely clear the tumor at an E:T ratio in which the CAR-T cells incubated with only the antibody alone were unable to control the tumor growth. Interestingly, combination therapy of CAR-T cells and antibody-cytokine fusions outperformed CAR-T cells constitutively secreting the same cytokines in in vitro assays. This work highlights the use of a single reagent capable of increasing the efficacy of the majority of CAR-T cell therapies and may be an important contribution towards treating solid tumor malignancies with CAR-T cell therapies.

Citation Format: John T. Keane, Avery D. Posey. PanCAR-specific antibody-cytokine fusion proteins. [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 4089.
Abstract 4141: Effects of exercise on tumorigenesis, immune system, and the gut microbiome in a bladder carcinogen mouse model
Authors: LaMorte Joseph P. , Tian Brittany , Ginwala Rashida , Bukavina Laura , Abbosh Philip
Published: April 2023

Recent studies have begun to demonstrate the therapeutic capacity of physical exercise on cancer treatment outcomes. While this research suggests exercise may improve health in cancer patients, the specific mechanisms by which exercise influences tumor progression remain largely unknown. We hypothesized that exercise alters the gut microbiome and promotes immune cell activation in relevant immune sites. Therefore, we utilized BBN (N-butyl-N-(4-hydroxybutyl)-nitrosamine), a carcinogen that mimics human muscle invasive bladder cancer conditions, and voluntary exercise wheels to determine the effect of exercise on bladder carcinogenesis, gut microbiome, and the immune system. Ultrasound imaging revealed that the average tumor volume of female exercising mice was 11.9% of the total bladder volume at time of sacrifice, while the tumor volume of their non-exercising counterparts was 56.4%, a difference determined to be significant using a two-tailed t-test with p-value of 0.05. A similar trend was seen in male mice, though the difference between groups was not significant. Immune cell analysis did not show significant variation in cytotoxic T cell, helper T cell, macrophage, or dendritic cell populations between exercising and non-exercising mice in spleen, colon, or bladder tissues as measured by FACS. Standardization of the amount exercised, as evaluated by expression of myosin and actin levels in the tibialis anterior muscle, and sequencing of the gut microbiome are in progress and could better indicate the mechanism of exercise-induced tumor suppression.

Citation Format: Joseph P. LaMorte, Brittany Tian, Rashida Ginwala, Laura Bukavina, Philip Abbosh. Effects of exercise on tumorigenesis, immune system, and the gut microbiome in a bladder carcinogen mouse model. [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 4141.
Abstract 4145: Frizzled-4 expression enhances hepatocellular carcinoma progression, sorafenib resistance and suppresses anti-tumor CD8 T cell immunity
Authors: Luo Jie , Gong Lanqi , Yang Yuma , Liu Qin , Huang Jiao , Fang Xiaona , Guan Xin-Yuan
Published: April 2023

Background: Hepatocellular carcinoma (HCC) is one of the most common and deadliest cancers. Sorafenib remains the globally accepted systemic first-line treatment for advanced HCC although acquired drug-resistance is becoming increasingly common. Hopefully, immune-checkpoint inhibitor therapy has recently revolutionized the treatment for advanced HCC that have poor response to sorafenib, but the durable response rate remains low in most patients for unknown reasons. This fuels a wave of research into the molecular mechanisms of tumor-intrinsic resistance to both chemotherapy and immune checkpoint blockade (ICB), which may provide potential systemic treatment options for advanced HCC patients.

Methods and Results: RNA sequencing of sorafenib-resistant HCC cell lines was performed to identify Frizzled-4 (FZD4) as a critical gene for drug resistance that was further validated in animal models. Clinical implication of FZD4 overexpression was studied in HCC patients. Using genetic and pharmacological approaches, we found that FZD4 was sufficient to increase clonogenicity, migration and invasion, and prevent sorafenib-induced apoptosis. In addition, FZD4 could reduce secretion of tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ), as well as intracellular levels of cytotoxic granzyme A and perforin in CD8+ T cells, indicating significant suppression of CD8+ T cell-mediated antitumor immunity in HCC tumors. In vivo studies also revealed that FZD4 expression promoted tumorigenesis and accelerated HCC cell metastatic lung colonization. Combination therapy of FZD4 inhibitor FzM1 with sorafenib in HCC of MYC/sgp53-induced model showed a tendency to significantly impede tumor growth and improve mice survival. Moreover, gene set enrichment analysis (GSEA) identified the cholesterol homeostasis and fatty acid metabolism signaling signatures as the top activated pathways in FZD4-high HCC cells. This evidence suggests a crucial role of FZD4-involved lipid metabolism linked to the development of therapeutic resistance.

Conclusion: Our study demonstrates that FZD4 expression plays a pivotal role in HCC progression and holds the potential for expanding the scope of targeted/ICB therapies to tumors that are currently unresponsive, especially in advanced HCC. The resistance mechanisms will be addressed in further work.

Citation Format: Jie Luo, Lanqi Gong, Yuma Yang, Qin Liu, Jiao Huang, Xiaona Fang, Xin-Yuan Guan. Frizzled-4 expression enhances hepatocellular carcinoma progression, sorafenib resistance and suppresses anti-tumor CD8 T cell immunity. [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 4145.