bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3929: Patient-derived pancreatic cancer organoids as in-vitro cancer models for personalizing therapy
Authors: Chew Claire Alexandra Zhen , Tay Li Min , Hooi Lissa , Chow Edward Kai-Hua , Bonney Glenn K. , Chan Shing Leng
Published: September 2020

Pancreatic ductal adenocarcinoma (PDAC) is the most aggressive solid tumor. Unfortunately, patients with this disease have not benefited from the advances in precision medicine seen in other malignancies. The lack of actionable mutations as well as predictive biomarkers has hampered progress in this area. Combination therapy remains the cornerstone of systemic treatment, with FOLFIRINOX being the most effective therapeutic regimen in PDAC. However, it is clear that only subset of patients are responsive, and the toxicities of this 4-drug combination are intolerable to many patients particularly in the adjuvant setting. There is a clear need for more personalized rational drug combinations that are optimized to maximize efficacy while minimizing toxicity.

We have established a protocol for the generation of PDAC organoids from endoscopic ultrasound biopsies for personalizing systemic therapy based on the principle of phenotype driven oncology. Organoids generated are analyzed for KRAS and p53 mutations to confirm their tumor origin. By applying a phenotypic optimization platform (QPOP) on these organoids, we are able to identify optimized drug combinations from a pool of both FDA-approved and preclinical drugs. As expected, FOLFIRINOX did not always rank high by treatment efficacy suggesting that alternative treatment may be more beneficial for these patients. In addition, we identified that the efficacy of FOLFIRINOX was driven by two-drug synergistic combinations which varied between patients. This can potentially advise modifications to this regimen when toxicity arises. From a discovery perspective, our platform has the capability of deriving novel drug combinations. The combination of gemcitabine and Abraxane with a WEE1-inhibitor was found to be synergistic in selected PDAC patients. Furthermore, patients with similar drug response profiles can be studied together to identify predictive markers of sensitivity.

To demonstrate that this platform can indeed be applied to prospectively recommend treatment strategies for patients with progressive disease on first-line chemotherapy, we are establishing a clinical trial to personalize second-line therapy, with the endpoint of overall response rate. Overall, by combining patient-derived organoids with rational optimized drug combinations, we aim to personalize therapy for patients with PDAC to improve overall survival.

Citation Format: Claire Alexandra Zhen Chew, Li Min Tay, Lissa Hooi, Edward Kai-Hua Chow, Glenn K. Bonney, Shing Leng Chan. Patient-derived pancreatic cancer organoids as in-vitro cancer models for personalizing therapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3929.
Abstract 3964: Integrin α2-collagen I axis promotes proliferative and migratory phenotypes in intrahepatic cholangiocarcinoma
Authors: Rattanasinchai Chotirat , Navasumrit Panida , Ruchirawat Mathuros
Published: September 2020

Cholangiocarcinoma (CCA) is a deadly cancer of the bile duct and is considered an incurable disease with no efficacious treatment option. CCA arises as a consequence of chronic inflammation and tissue damage to the biliary epithelium. Thus, one of the crucial characteristics of CCA is its tumor microenvironment high in stromal extracellular matrices (ECMs), especially collagen I. Herein, the direct impact of collagen I on intrahepatic CCA (iCCA) cell lines were investigated. Our data demonstrate that iCCA cells mainly interact with collagen I, while cholangiocyte cells preferentially binds to fibronectin in cell adhesion assays. Correspondingly, iCCA cell lines exhibit over 20 fold-upregulation of ITGA2 gene, which encodes collagen-binding integrin α2, when compared with non-cancerous cholangiocyte cells. Furthermore, the result from the GEO dataset of 92 iCCA patients demonstrate that ITGA2 gene expression in iCCA tumors is 13 folds greater than their adjacent normal tissues and is positively correlated with expressions of two isoforms of collagen I gene, COL1A1 and COL1A2. Collagen I is also sufficient to drive proliferation of iCCA cells in a serum-restricted condition and this effect is mediated by integrin α2 as proven by ITGA2 silencing or α2β1-selective inhibitor treatment. In contrast to round-shaped, multicellular colonies of iCCA cells in clonogenic assays, iCCA colonies exhibit migratory phenotypes in the presence of collagen I. Likewise, this phenotype can be inhibited by α2β1-selective inhibitor treatment. Taken together, these data suggest that stromal collagen I could be one of the tumor microenvironmental factors directly facilitating iCCA progression. Thus, targeting the integrin α2-collagen I axis could be a promising therapeutic strategy for combating this cancer.

Citation Format: Chotirat Rattanasinchai, Panida Navasumrit, Mathuros Ruchirawat. Integrin α2-collagen I axis promotes proliferative and migratory phenotypes in intrahepatic cholangiocarcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3964.
Abstract 3985: GDF15 expression level impacts the canonical pathways of the tumor microenvironment and ovarian tumors with wild-type p53 in response to chemotherapy
Authors: Izaguirre Daisy I. , Tsang Yvonne TM , Kun Eucharist H. , Gershenson David M. , Wong Kwong-Kwok
Published: September 2020

Background

The standard treatment for ovarian cancer is surgery, followed by chemotherapy with a platinum agent, such as cisplatin or carboplatin, and a taxane, such as paclitaxel. We previously observed that patients with ovarian cancer that was wild-type for p53 had a lower survival rate than did those with p53 mutations. A better understanding of the molecular changes that occur in epithelial ovarian cancer cells with wild-type p53 in response to cisplatin treatment could reveal a novel mechanism of chemoresistance. Therefore, we might be able to develop novel strategy to overcome chemoresistance.

Methods

Gene expression profiling was performed on the wild-type p53 ovarian cancer cell line A2780 to identify genes that are highly induced by cisplatin. Growth differentiation factor 15 (GDF15) was identified as being highly induced by cisplatin or carboplatin. This finding was validated in a panel of wild-type and mutant p53 ovarian cancer cell lines. Mouse orthotopic models were used to further investigate the induction of GDF15 by cisplatin in vivo and the effect of GDF15 knockdown (GDF15-KD) on tumor growth and cisplatin response. A novel analysis was performed of RNAseq data generated from tumors formed in cisplatin-treated orthotopic mouse models to understand the tumor and stromal cell signaling changes.

Results

The induction of GDF15 was demonstrated in vivo, and the serum level of GDF15 after cisplatin treatment was also found to be significantly up-regulated in an A2780 ovarian cancer orthotopic mouse model. More importantly, cisplatin treatment was more effective in shrinking the mouse tumors developed from GDF15-KD A2780 cell line than did control tumors. Histopathological and novel RNAseq analyses of mouse tumors revealed that GDF15-KD tumors had a lower stromal component than did control tumors after cisplatin treatment. GDF15-KD tumors also had a different repertoire of activated and inhibited canonical pathways in their stromal cell and cancer cell components than did control tumors in response to cisplatin treatment.

Conclusions

GDF15 expression in wild-type p53 ovarian cancer cells can modulate the canonical pathways in the tumor microenvironment in response to cisplatin and may be a mechanism of chemoresistance. Targeting GDF15 signaling could be a novel strategy to enhance the effectiveness of chemotherapy.

Citation Format: Daisy I. Izaguirre, Yvonne TM Tsang, Eucharist H. Kun, David M. Gershenson, Kwong-Kwok Wong. GDF15 expression level impacts the canonical pathways of the tumor microenvironment and ovarian tumors with wild-type p53 in response to chemotherapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3985.
Abstract 4034: High throughput synergy screening identifies therapeutic combinations for Merkel cell carcinoma
Authors: Garman Khalid , Gelb Tara , Urban Daniel , Hall Matthew , Brownell Isaac
Published: September 2020

Merkel cell carcinoma (MCC) is a rare and aggressive neuroendocrine skin cancer. MCC is virus-positive (VP-MCC) in the majority of cases wherein Merkel cell polyomavirus is integrated into the host genome. The remainder of cases are virus-negative (VN-MCC) and are associated with mutations induced by ultraviolet light. Primary MCC is effectively treated by surgical resection and radiotherapy, but for metastatic MCC, outcomes for conventional chemotherapy are poor. Immune checkpoint inhibitors are more likely to produce durable responses in metastatic MCC, but they are not effective in all cases and are not a viable option for patients with immunosuppression. To address the need for better therapeutic options, we utilized high-throughput small molecule screening to assess the efficacy of ~4,000 drugs in reducing VP-MCC and VN-MCC viability. Among the compounds we screened, navitoclax was identified as an effective agent against VP-MCC cell lines, compared to VN-MCC or control cells. However, navitoclax clinical application is limited by dose-dependent thrombocytopenia. One strategy to reduce thrombocytopenia risk is to combine a lower dose of navitoclax with another MCC-effective agent. To identify effective drug combinations, we performed a synergy screen with 1912 pairings of navitoclax with each of the clinically relevant compounds in the NCATS Mechanism Interrogation PlatE (MIPE). We were able to identify synergy between navitoclax and multiple drug classes. High priority drug combinations are now being evaluated for toxicity and efficacy in pre-clinical xenograft models of MCC. Our study has identified novel therapeutic combinations for the treatment of VP-MCC. By focusing on drugs already in clinical use, validated combinations will be quickly available to patients for whom immunotherapy is not available or effective.

Citation Format: Khalid Garman, Tara Gelb, Daniel Urban, Matthew Hall, Isaac Brownell. High throughput synergy screening identifies therapeutic combinations for Merkel cell carcinoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4034.
Abstract 4063: Design and development of a novel 5-FU modified siRNA against BCL2 with enhanced efficacy and the unique ability to be delivered vehicle free into cancer cells
Authors: Fesler Andrew , Hwang Ga-Ram , Yuen John , Ju Jingfang
Published: September 2020

Evasion of apoptosis is one of the major hallmarks of cancer and an important factor in chemoresistance. Thus, targeting apoptosis regulation is an important therapeutic approach. Anti-apoptotic BCL2, an oncogene and therapeutic target, is overexpressed in several different cancer types. There have been various approaches developed to target BCL2 including the FDA approved Venetoclax. In this study we have developed a modified siRNA against BCL2 that has enhanced efficacy and also can be delivered into cancer cells without any vehicle. We developed this siRNA by replacing uracil residues in the siRNA with 5-flurouracil (5-FU). This modification incorporates the effects of both 5-FU and siBCL2, to have an enhanced therapeutic effect. We tested several different modified siBCL2's, one with only the sense strand modified, one with only the antisense strand modified and one with both strands modified. We found the modified siBCL2 with both strands modified with 5-FU was most effective at inhibiting BCL2. Importantly, we were able to demonstrate via qRT-PCR and Western blot, that this modified siBCL2 retains its siRNA function and inhibits BCL2 expression in several different cancer types including colon cancer, lung cancer and lymphoma. In addition, in lymphoma cells, we found that our modified siBCL2 has better efficacy than BCL2 inhibitor Venetoclax. Finally, we were able to demonstrate that modified siBCL2 can be delivered into cancer cells without a delivery vehicle. This unique feature may be significant in overcoming the challenge of delivery, which remains a major obstacle for RNA therapeutics. This modified siBCL2 has therapeutic potential for several different cancers. Beyond BCL2, the modification approach we demonstrated may be applicable to other siRNAs and has potential as a platform technology for siRNA based therapeutics.

Citation Format: Andrew Fesler, Ga-Ram Hwang, John Yuen, Jingfang Ju. Design and development of a novel 5-FU modified siRNA against BCL2 with enhanced efficacy and the unique ability to be delivered vehicle free into cancer cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4063.
Abstract 4084: A DGKA-cJUN-WEE1 signaling axis provides platinum resistance in ovarian cancer
Authors: Li Jie , Pan Chaoyun , Boese Austin C. , Umano Anna , Kang Sumin
Published: September 2020

Although platinum compounds are the first-line treatment for ovarian cancer, the majority of patients relapse and develop resistance to treatment. However, the mechanism underlying resistance is unclear. Here, we report a unique platinum resistance mechanism that is mediated by a lipid messenger, phosphatidic acid (PA), in ovarian cancer. A phospho-proteomics screen revealed that PA, which is predominantly produced by diacylglycerol kinase alpha (DGKA) in ovarian cancer, activates the transcription factor c-JUN. DGKA and PA facilitate JNK recruitment to c-JUN and nuclear localization leading to its activation upon cisplatin exposure. Through a genomic screen, we identified that activated c-JUN enhances expression of WEE1 and consequently prevents cells from aberrant cell cycle progression upon cisplatin exposure. DGKA-cJUN-WEE1 signaling was elevated in tumors derived from ovarian cancer patients who did not respond to platinum-based therapy compared to tumors from patients who responded to treatment. Our study demonstrates how lipid metabolism, kinase signaling, and gene regulatory systems are intertwined to direct cisplatin resistance in ovarian cancer.

Citation Format: Jie Li, Chaoyun Pan, Austin C. Boese, Anna Umano, Sumin Kang. A DGKA-cJUN-WEE1 signaling axis provides platinum resistance in ovarian cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4084.
Abstract 4085: Blocking Fra-1 sensitises triple-negative breast cancer to olaparib
Authors: Song Dandan , He Huan , Sinha Indranil , Pettersson Linnea , Yan Feifei , Haldosen Lars-Arne , Zhao Chunyan , Williams Cecilia
Published: September 2020

Fra-1 (FOSL1), a member of the activator protein 1 (AP-1) transcription factor complex, is overexpressed in triple-negative breast cancer (TNBC) and plays crucial roles in tumor progression and treatment resistance. We have previously identified 118 proteins that interact with endogenous chromatin-bound Fra-1 in TNBC cells in a large screen, and this included PARP1 (Poly (ADP-ribose) polymerase 1). PARP1 inhibitor olaparib is currently in use for treatment of BRCA-mutated TNBC breast cancer. Here, we corroborate that PARP1 interacts with Fra-1, and we also find that PARP1 downregulates Fra-1 and reduces AP-1 transcriptional activity. Inhibition of PARP1, on the other hand, increases Fra-1 levels and enhances AP-1 transcriptional activity. Further, we find that upon inhibition of Fra-1, TNBC cells become sensitized to olaparib treatment. By comparing the Fra-1 and PARP1 regulated transcriptomes with Fra-1 chromatin binding site, we determine that a large fraction of PARP1-regulated genes is dependent on Fra-1. Finally, we show that PARP1 protein levels significantly correlate with Fra-1 in clinical breast cancer tumors, and we identify that while high PARP1 expression is indicative of a poor clinical outcome in breast cancer patients overall, it is not in basal-like tumors. In conclusion, by exploring the functionality of the Fra-1 and PARP-1 interaction, we propose that targeting Fra-1 could serve as a therapeutic approach to improve olaparib treatment outcome for TNBC patients.

Citation Format: Dandan Song, Huan He, Indranil Sinha, Linnea Pettersson, Feifei Yan, Lars-Arne Haldosen, Chunyan Zhao, Cecilia Williams. Blocking Fra-1 sensitises triple-negative breast cancer to olaparib [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4085.
Abstract 4089: Investigating translesion synthesis DNA polymerases roles on tumor protection after temozolomide induced DNA damage
Authors: Latancia Marcela Teatin , Bastos André Uchimura , Moreno Natália Cestari , Jardim Davi , Rocha Clarissa RR , Menck Carlos FM
Published: September 2020

Translesion DNA polymerases are capable of replicating damaged DNA, without removing the lesions, performing translesion synthesis (TLS), a mechanism of DNA damage tolerance. Tumor cells use this mechanism in order to survive lesions caused by chemotherapy and, therefore, this may be a strategy that tumor cells use to resist treatments. Moreover, it is an error-prone process and can lead to mutagenesis increasing resistance potential of tumor cells. Little is known about the role of TLS in Temozolomide (TMZ) tumor treatment. Our aim is to investigate how TMZ affects TLS mutated cells. Hence, we firstly treated primary fibroblasts cells lacking TLS mechanisms (XP-V) and analyzed cell proliferation by flow cytometry, cell viability by a colorimetric assay (XTT), for survival by apoptotic markers (such sub-G1) and clonogenic assays. In fact, XP-V cells are more sensitive to TMZ, indicating that TLS mechanisms are important to overcome DNA damage. TCGA data was also used to analyze TLS expression in glioblastoma patients. Curiously, in recurrently patients treated with TMZ, POLK expression was significantly increased. CRISPR/CAS9 knockout (KO) cells of glioblastoma cell lines, such as U251, were constructed for other TLS DNA polymerases enzymes genes, such as Pol Iota and Pol kappa. Gene KO was validated using Sanger sequencing and mutations resulted in stop codon. These pol iota and pol kappa KO cells had impaired viability after TMZ treatment. Moreover, genotoxic stress, as indicated by phosphorylated gH2AX, were increased in TMZ-treated cells. In summary, we can infer that TLS polymerases protect tumor cells from TMZ-induced DNA damage and thus, play important roles on overcoming resistance to this drug. As perspectives for this work, we intend to observe how replication is affected in these KO cells treated with TMZ by DNA Fiber Assays and measure DNA strand breaks in those cells by comet assay. Financial support: FAPESP, CNPq and CAPES.

Citation Format: Marcela Teatin Latancia, André Uchimura Bastos, Natália Cestari Moreno, Davi Jardim, Clarissa RR Rocha, Carlos FM Menck. Investigating translesion synthesis DNA polymerases roles on tumor protection after temozolomide induced DNA damage [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4089.
Abstract 4144: A tissue-microfluidic derived aptamer displays prognostic and therapeutic characteristics in serous ovarian carcinoma through heat shock protein 70(HSP70)
Authors: Lin Chang-Ni , Liang Yu-Ling , Wu Pei-Ying , Lee Gwo-Bin Vincent , Hsu Keng-Fu
Published: September 2020

Background: Previously, we have identified aptamers through ovarian cancer (OC) tissue microfluidic platform which show high specificity to ovarian cancer tissue and cells. However, the clinical diagnostic and therapeutic roles of these aptamers are still largely unknown.

Materials and Methods: Liquid chromatography-mass spectrometry (LC-MS) was used to find binding candidates of aptamer, Tx-01, from serous type ovarian cancer cells. Immunohistochemistry (IHC), clinical survival analysis, western blotting and confocal microscopic images were performed to detect the expression of Tx-01 and HSP70/ Notch1 intracellular domain (NICD) in OC tissues. The functions of Tx-01 were studied in vitro and in vivo experiments. Inhibitors treatment, nuclear protein extraction and western blotting were carried out to demonstrate the possible mechanisms of Tx-01 function.

Results: High expression Tx-01 in OC tissues was correlated to poor patients' survival in serous ovarian carcinoma. In vitro experiments showed that Tx-01 has a feature of recognizing membrane-bound HSP70. In the meantime, Tx-01 inhibit HSP70 signal activation via interrupting intracellular HSP70/NICD interaction to reduce cell migration and invasion. Furthermore, Tx-01 inhibited the tumor growth in the mice experiment.

Conclusion: Our data showed that Tx-01 acted as a prognostic predictor in serous OC through recognizing the membrane-bound HSP70. Also, Tx-01 reduced the tumor cell migration, invasion through blocking HSP70 signal activation. Thus, our work demonstrated that Tx-01 might be a prognostic biomarker and potential inhibitor in serous OC therapy.

Citation Format: Chang-Ni Lin, Yu-Ling Liang, Pei-Ying Wu, Gwo-Bin Vincent Lee, Keng-Fu Hsu. A tissue-microfluidic derived aptamer displays prognostic and therapeutic characteristics in serous ovarian carcinoma through heat shock protein 70(HSP70) [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4144.
Abstract 417: Developing mouse models to dissect the function of a driver gene epimutation in lung tumorigenesis
Authors: Yang Li , Mandala Anusha , Chen Yi , Shen Lanlan
Published: September 2020

Background: Cancer is a complex disease caused by the accumulation of both genetic and epigenetic alterations in oncogenes and tumor suppressor genes. Although an extensive catalog of epigenetic changes has been generated in patients with lung cancer, we have only a fragmentary understanding of their functional roles, particularly for the initiation and maintenance of tumors. The use of genetically engineered mouse models definitively demonstrate that mutations at specific genes cause cancer. An analogous “epigenetically engineered mouse models” should not only lead to important insights about how cancer develop but could provide in vivo platforms in which to screen and validate novel epigenetic therapies.

Methods and Results: We published the first epigenetically engineered mouse cancer model based on a driver gene epimutation of p16 (also known as cyclin-dependent kinase inhibitor 2A). In human lung cancers, p16 epimutation is common, occurring in ~50% of cases. Importantly, p16 epimutation is frequently detected in lung pre-neoplastic lesions and p16 is of the best documented gene that controls K-RAS oncogene-induced senescence. Therefore, to elucidate the functional role of p16 epimutation for lung cancer establishment and progression, we developed a mouse model that combines oncogenic K-ras with p16 epimutation. We found that mice with combined K-ras mutation and p16 epimutation (K-ras+/G12D; p16meth/meth) displayed substantially increased tumor numbers and sizes, with tumor progression to adenocarcinoma. In addition, to investigate the therapeutic effects of reversing p16 epimutation, we developed a mouse model that enables conditional and inducible CRISPR based epigenetic gene editing. Our system utilizes deactivated Cas9 (dCas9) fused to a repetitive protein scaffold known as SunTag, which recruits multiple copies of antibody-fused demethylase TET1. We showed that this system is robust and reproducible to achieve targeted DNA demethylation at CpGs within 100 bp from the sgRNA target site of p16 promoter and to reactivate p16.

Conclusion: We have established clinically relevant and immunocompetent mouse models based on defined genetic and epigenetic alterations to drive benign lung tumor growths towards the ultimate malignant phenotype. Our model should prove useful as an in vivo preclinical platform for testing epigenetic therapeutic strategies.

Citation Format: Li Yang, Anusha Mandala, Yi Chen, Lanlan Shen. Developing mouse models to dissect the function of a driver gene epimutation in lung tumorigenesis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 417.
Abstract 4170: Decursin inhibits gastric cancer cell growth and autophagic flux by suppressing cathepsin C
Authors: Kim Solbi , Joo Mina , KIM Nayoung , Lee Myung-Won , Jeon Heung Jin , Ryu Hyewon , Song Ik-Chan , Lee Hyo Jin
Published: September 2020

Purpose: Autophagy is a series of processes in which biomolecules are digested and recycled, and it also provides energy for the development and progression of cancer. Thus, autophagy inhibitors can be used to suppress the progression of cancer. In this study, we investigated whether decursin, which is derived from natural products, inhibits the growth and autophagy of cancer cells.

Methods: The gastric cancer cell lines SNU216 and NCI-N87 were treated with or without decursin. Cell growth was evaluated by Cell-Counting Kit-8, colony formation, and BrdU assays, as well as by propidium iodide staining, and autophagic flux was evaluated by western blotting, LC3 puncta, GFP-RFP-LC3, and transmission electron microscopy. Finally, tumor spheroids and patient-derived gastric cancer organoids were used to confirm the effect of decursin in three-dimensional culture.

Results: Decursin reduced the growth and anchorage-dependent viability of gastric cancer cells. LC3 cleavage and LC3-GFP puncta were increased by decursin in a time- and concentration-dependent manner. However, the increase in autophagosome formation did not lead to an increase in autophagic flux. Decursin downregulated cathepsin C (CTSC), a lysosomal protease, irrespective of cellular pH. In addition, knockdown of CTSC reduced the mRNA and protein levels of E2F3, resulting in cell-cycle arrest. Decursin also reduced cell growth in both tumor spheroids and patient-derived gastric cancer organoids.

Conclusion: The results demonstrated that decursin inhibited tumor growth and progression by suppressing CTSC-mediated autophagic flux and cell-cycle progression. Therefore, decursin is a novel inhibitor of autophagic flux, with potential for the treatment of gastric cancer.

Citation Format: Solbi Kim, Mina Joo, Nayoung KIM, Myung-Won Lee, Heung Jin Jeon, Hyewon Ryu, Ik-Chan Song, Hyo Jin Lee. Decursin inhibits gastric cancer cell growth and autophagic flux by suppressing cathepsin C [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4170.
Abstract 4177: Preclinical development of ABM-1310, a novel BRAF Inhibitor to treat cancer with brain metastasis
Authors: Hu Yong , Chen Chen , Huang Charles , Xu Min , Chen Youqin , Chen Xiyuan , Zhao Li
Published: September 2020

BRAF is a key molecule activated by RAS in the MAPK pathway that mainly controls cell proliferation. BRAF mutation is detected in about 8% of all cancer population, and more than 50% is BRAFv600e, which causes constitutive MAPK activation. BRAFv600 mutation is found in various cancers such as melanoma, colorectal cancer, glioma, lung cancer, thyroid cancer, pancreatic cancer, ovary cancer etc.

ABM-1310 is a novel small molecule BRAF inhibitor that specific inhibits BRAFv600 mutant form with high water solubility, cell permeability, and blood brain barrier (BBB) penetration. Although current target or immune-therapies are reported with some variable effects, melanoma with brain metastasis is still a significant challenge in clinic. With high BBB penetration and high exposure in brain, ABM-1310 has good potential to treat tumors bearing BRAFv600 mutation with brain metastasis.

In-vitro data shown similar on-target enzyme activity of ABM-1310 to marketed BRAF inhibitors on BRAFv600e, as well as high anti-proliferation activity in multiple cancer cell lines with BRAF V600E mutation like A375, Colo-829, and HT-29 etc. In vivo pharmacology studies demonstrated that ABM-1310 had good potency of tumor growth inhibition with oral dose as low as 1mg/kg/day. ABM-1310 treatment also improved animal overall survival in several cell-line derived xenograft (CDX) models, including subcutaneous, intracranial and intracardiac melanoma models. Non-clinical pharmacokinetics studies also shown ABM-1310 had a favorable ADME profile both in vitro and in vivo in animals. ABM-1310 demonstrated excellent brain penetration in animals.

Single-dose, seven-day repeat dose non-GLP studies and four-week GLP toxicity studies in SD rats and beagle dogs all revealed that ABM-1310 had a good safety profile with NOAEL of 100mg/kg/day in SD rats and 30mg/kg/day in beagle dogs.

Based on these supportive preclinical study results, IND of ABM-1310 was submitted in 2019 Q4 to investigate its safety in humans as signal reagent, which will be followed by further clinic development of ABM-1310, or combination with other molecules to treat cancers with BRAFv600 mutation, especially with brain metastasis.

Citation Format: Yong Hu, Chen Chen, Charles Huang, Min Xu, Youqin Chen, Xiyuan Chen, Li Zhao. Preclinical development of ABM-1310, a novel BRAF Inhibitor to treat cancer with brain metastasis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4177.
Abstract 4279: Automated TRB locus haplotype analysis by long-amplicon TCRB chain sequencing for immune-related adverse events biomarker research
Authors: Burke Jennifer , Chan Frances , Huang Jiajie , Sedova Marina , Looney Timothy , Hyland Fiona
Published: September 2020

Introduction: Identifying predictive biomarkers for immune related adverse events (irAEs) following checkpoint blockade inhibition (CPI) remains an outstanding goal of immune-oncology translational research. Polymorphism within the T cell receptor variable gene (TRBV) has been proposed as a potential risk factor for irAEs owing to the potential link between TRBV polymorphism and chronic autoimmune disease. Efforts to interrogate the potential biomarker utility of TRBV polymorphism have been hampered by the repetitive nature of the TRB locus. We previously demonstrated a method for inferring TRB locus haplotypes from long amplicon TCRB chain sequencing data, which we used to identify major haplotype groups in a set of 81 Caucasians. Here we present an automated method for haplotype group assignment from TCRB chain sequencing data.

Methods: Rearranged TCRB chains from 10 blood samples were amplified and sequenced from 25ng peripheral blood total RNA via the Oncomine™ TCRB-LR assay using the Genexus™ Integrated Sequencer. 12 samples were run per chip with 4 samples run in each lane. TCRB clonotyping and repertoire feature analysis was performed using the Genexus™ software v5.107.14. Automated haplotype group assignment was performed by generation and comparison of TRBV allele profiles to those presented in DOI: 10.4103/JIPO.JIPO_16_19. For context, TCR evenness, convergence, and haplotype group assignment were compared to values obtained following analysis of the same samples via the GeneStudio™ S5 platform and Ion Reporter™ 5.12 software.

Results: TCR Evenness and convergence values were highly correlated across replicates run on the Genexus™ Integrated Sequencer (Spearman correlation >0.95 and >.70, respectively). Evenness at equivalent clone count and convergence at equivalent sequencing depth were not significantly different across platforms (Spearman correlation >0.88). Haplotype group assignments demonstrated 100% agreement across replicates on both platforms.

Conclusions: We demonstrate an automated and reproducible research method for TRB haplotype group assignment via the Oncomine™ TCR-Beta LR Assay, GX run on the Genexus™ Integrated Sequencer. Future studies will evaluate the potential biomarker utility of TRB haplotyping for the prediction of irAEs.

Citation Format: Jennifer Burke, Frances Chan, Jiajie Huang, Marina Sedova, Timothy Looney, Fiona Hyland. Automated TRB locus haplotype analysis by long-amplicon TCRB chain sequencing for immune-related adverse events biomarker research [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4279.
Abstract 4298: The gut microbiome is associated with clinical responses to anti-PD-1 immunotherapy in gastrointestinal cancer patients in China
Authors: Peng Zhi , Cheng Siyuan , Zhang Quan , Kou Yan , Wang Ziqi , Jin Rong , Hu Han , Zhang Xiaotian , Gong Jifang , Li Jian , Lu Ming , Wang Xicheng , Zhou Jun , Lu Zhihao , Bi Dongtao , Tan Yan , Shen Lin
Published: September 2020

We report the first and largest cohort, to the best of our knowledge, on the gut microbiome of 126 gastrointestinal (GI) cancer patients in China receiving the immune checkpoint inhibitors (ICI) targeting PD-1. The cohort included 31 patients with colon cancer, 23 with esophageal carcinoma, 47 with gastric cancer, and 25 with other cancer types. 43 of the patients had samples collected before treatment. 35 of patients were assessed as microsatellite-instability-High (MSI-High) and 71 as non-MSI-High. 408 fecal samples from the patients were collected. We performed 16s rRNA amplicon sequencing (V4 region) of the patients' samples to investigate the relationship between microbial profiles and patients' immunotherapy treatment response (RECIST 1.1). Principal coordinate analysis (PCoA) showed a subgroup of responders exhibiting a microbiome profile which was enriched in Prevotella. Hierarchical clustering of patients' microbiome profiles also suggested that for each gastric and colon cancer, patients with the same response tend to share similar microbiome profiles. By comparing the microbiome profiles between R and NR group using samples collected before treatment or within 3 weeks since the start of treatment, we identified 16s rRNA V4 sequences from Prevotella enriched in R group for each tumor type, which suggests positive association between Prevotella and tumor response. Our data also suggested that MSI as good marker for tumor response, although more data is required to achieve the significance level. Besides, no significant difference in alpha diversity was observed between R and NR group for each tumor type. In sum, we studied the microbiome profiles of 126 GI cancer patients and evaluated their association with anti-PD-1 immunotherapy. Patients with the same tumor type and response tend to share similar microbiome profiles, where Prevotella and a few other taxa contributed to the pattern.

Citation Format: Zhi Peng, Siyuan Cheng, Quan Zhang, Yan Kou, Ziqi Wang, Rong Jin, Han Hu, Xiaotian Zhang, Jifang Gong, Jian Li, Ming Lu, Xicheng Wang, Jun Zhou, Zhihao Lu, Dongtao Bi, Yan Tan, Lin Shen. The gut microbiome is associated with clinical responses to anti-PD-1 immunotherapy in gastrointestinal cancer patients in China [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4298.
Abstract 4302: Molecular profile of KRAS G12C-mutant non-small-cell lung cancer and colorectal cancer in Brazilian patients
Authors: Souza Bianca Mendes , Leite Laura Rabelo , Parma Sabrina A. , Lopes Natália P. , Malta Frederico S. , Araujo Luiz Henrique , Freire Maíra C.
Published: September 2020

Background: Cancer is one of the leading causes of death and one of the main barriers to increasing life expectancy worldwide. The development of therapeutic strategies directly targeting mutant oncogenes, such as EGFR and BRAF, has resulted in substantial benefits for patients with a variety of cancers. KRAS is one of the most frequently mutated oncogenes, however efforts to develop targeted therapies have been largely unsuccessful. Recently, two small-molecule inhibitors, AMG 510 and MRTX849, have shown promising activity in KRAS G12C-mutant solid tumors. The current study aims to assess the molecular profile of KRAS G12C in non-small-cell lung cancer (NSCLC) and colorectal cancer (CRC) from Brazilian patients tested in a clinically-certified laboratory.

Methodology: 4,842 NSCLC patients were tested for KRAS, NRAS, EGFR and BRAF mutations by next generation sequencing (NGS), while 4,815 CRC patients were tested for KRAS and NRAS mutations, by pyrosequencing, between 2017 and 2019

Results and Discussion: From the 4,842 NSCLC samples tested, KRAS G12 mutations were identified in 1013 cases (20.9%) and, 336 of these (6.9% of total) were KRAS G12C. Additionally, from 4,815 CRC samples tested, KRAS G12 mutations were identified in 1633 cases (33.9%), and 166 of these (3.4% of total) were KRAS G12C. In the literature, KRAS G12C has been reported in 14 to 16% of all lung adenocarcinomas, and 5% of colorectal adenocarcinomas. Our data illustrate a relevance of this mutation in a significant number of patients and highlight an opportunity for targeted therapy. Some studies characterizing lung cancers have indicated that KRAS mutations, including KRAS G12C, are mutually exclusive with other known oncogenic driver mutations in NSCLC. However, in the current analysis, we identified 30 cases of NSCLC harboring multiple mutations, such as KRAS G12C mutations co-existing with EGFR, BRAF or NRAS mutations. This finding is probably due to the multiclonal presentation in selected cases.

Conclusions: A relatively high number of patients may present with KRAS G12C-mutant cancers in Brazil, indicating the relevance of identifying such cases for targeted therapy. Further studies will deepen the analysis of clonality and heterogeneity in this cohort.

Citation Format: Bianca Mendes Souza, Laura Rabelo Leite, Sabrina A. Parma, Natália P. Lopes, Frederico S. Malta, Luiz Henrique Araujo, Maíra C. Freire. Molecular profile of KRAS G12C-mutant non-small-cell lung cancer and colorectal cancer in Brazilian patients [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4302.
Abstract 4309: A novel immune prognostic index for the stratification of high-risk patients with early breast cancer
Authors: Lee Hannah , Kwon Mi Jeong , Koo Beom-Mo , Han Jinil , Shin Young Kee
Published: September 2020

The prognostic or predictive value of current commercial multigene assays for breast cancer are limited to hormone receptor-positive, human epidermal growth factor receptor 2-negative (HR+/HER2-) early breast cancer. Despite the prognostic or predictive significance of immune response-related genes in breast cancer, immune gene signatures have not been incorporated into most current assays. Here, we developed a novel immune gene based-prognostic model to predict the risk of recurrence in high-risk women with early breast cancer. Using public gene expression microarray datasets for breast cancer, we classified breast cancer patients into three risk groups according to clinical risk and proliferation risk, and identified the most significant five immune response-related genes (TRAT1, IL2RB, CTLA4, IGHM and IL21R) associated with clinical outcome in the clinical and proliferation high-risk (CPH) subgroup. Immune prognostic index based on the combination of the expression of five immune response-related genes and lymph node status was developed to predict the risk of recurrence in high-risk breast cancer patients and its prognostic significance was validated in independent validation datasets. Disease-free survival (DFS) or distant metastasis-free survival (DMFS) of patients classified as high risk according to the immune prognostic index was significantly lower than those of patients classified as intermediate and low risk in the CPH group. Multivariate analysis demonstrated that the immune prognostic index is an independent prognostic factor for DFS or DMFS. Moreover, c-index showed that the immune prognostic index is superior to clinicopathological variables in predicting the prognosis. Our immune index was also prognostic in independent validation datasets. Our immune prognostic index identified low-risk patients among patients classified as CPH group in all molecular subtypes of breast cancer and may be used to overcome the limitations of the current multigene assay.

Citation Format: Hannah Lee, Mi Jeong Kwon, Beom-Mo Koo, Jinil Han, Young Kee Shin. A novel immune prognostic index for the stratification of high-risk patients with early breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4309.
Abstract 4342: Histological variants and sites of presentation of malignant melanoma in a resource poor setting: A histopathological review
Authors: Adoke Kasimu Umar , Faruk Mohammed , Iliyasu Yawale , Ahmed Saad Aliyu
Published: September 2020

Background: Melanoma is a malignant tumour that arises from melanocytic cells. The incidence is increasing worldwide in white population where fair skin people receive excessive sun exposure. Although relatively uncommon in Africa-Americans, recent trends shows increase incidence in Africa- Americans. Data on the incidence and biology of melanoma are few in Africa. This study review the variants and sites of melanoma in a resource poor setting in Nigeria with some immunohistochemical stains performed.

Method: All cases of melanoma diagnosed in the Department of Pathology in a tertiary hospital in Nigeria over a three year period (2016-2018) were retrieved. Patient's information including site and age were also retrieved from their case notes. Immunohistochemistry using two antibodies S-100, HMB-45 was done.

Results: A total 38 cases were seen during the study period with a male female ratio of 1:1.1. The age range was 5-80 years with a mean age of 48.8±18.1. The most common site for melanoma is the foot 19(50%), followed by the uvea 7(18.4%), skin and inguinal lymph node 4(10.5%), anorectal and conjunctiva had 2(5.3%). The commonest histological variant of melanoma was acral lentigenous melanoma 19(50%), Superficial spreading melanoma 4(10.5%), nodular melanoma 3(7.9%) and desmoplastic melanoma 1(2.6%). Immunohistochemistry was used on three cases to rule out differentials.

Conclusion: Malignant melanoma in our setting is in contrast with African-Americans and Caucasians population base on site and histological type, hence future studies on molecular pathways of melanoma in Africa needs to be explored to show similarity or difference and the possibility of targeted immunotherapy.

Citation Format: Kasimu Umar Adoke, Mohammed Faruk, Yawale Iliyasu, Saad Aliyu Ahmed. Histological variants and sites of presentation of malignant melanoma in a resource poor setting: A histopathological review [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4342.
Abstract 435: Mouse models of human cancer
Authors: Bult Carol J. , Krupke Debra M. , Neuhauser Steven B. , Richardson Joel E. , Sundberg John P.
Published: September 2020

Mouse models play a crucial role in understanding the genetic and genomic basis of cancer and for the development of novel cancer therapies. Because data and information about these models are distributed and heterogeneous, gaining an appreciation for which models have been developed and what their characteristics are presents a major challenge for cancer researchers. The Mouse Models of Human Cancer Database (MMHCdb; http://tumor.informatics.jax.org) solves this challenge by providing a comprehensive, expertly curated on-line information resource about genetically engineered (GEMMs) and inbred mouse models of human cancer. In partnership with the European Bioinformatics Institute (EMBL/EBI), MMHCdb has also developed and implemented PDX Finder, a global catalog of Patient Derived Xenograft models (https://www.pdxfinder.org). Both MMHCdb and PDX Finder employ metadata standards to simplify the process of searching for relevant mouse models by diverse criteria including cancer type, type of strain, and genetic alteration. MMHCdb contains more than100,000 records on tumor frequency data for over 7,600 different mouse strains. These data are linked to over 4,700 references, 2,900 pathology reports, and 6,900 images. PDX Finder provides researchers with one stop access to information about more than 2,800 PDX models in repositories from around the world. MMHCdb and PDX Finder are supported, in part, by NIH/NCI CA089713. MMHCdb was formerly called the Mouse Tumor Biology database (MTB).

Citation Format: Carol J. Bult, Debra M. Krupke, Steven B. Neuhauser, Joel E. Richardson, John P. Sundberg. Mouse models of human cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 435.