bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2513: FGFRL1 deficiency reduces motility and tumor growth of cells derived from esophageal squamous cell carcinoma
Authors: Takei Yoshinori , Matsumura Takafumi , Watanabe Kazuaki , Nakamine Hirokazu , Sudo Tetsuo , Shimizu Kazuharu , Shimada Yutaka
Published: August 2018

Fibroblast growth factor receptor-like 1 (FGFRL1) belongs to the FGFR protein family. We previously reported that FGFRL1 is highly expressed in esophageal squamous cell carcinoma (ESCC) patients with lymph node metastasis and with the depth of ESCCs. Furthermore, transient inhibition of FGFRL1 expression induces cell cycle arrest and apoptosis of ESCC cells. However, the intracellular domain of FGFRL1 lacks the tyrosine kinase domain. Thus the mechanisms of which FGFRL1 contributes to the development of ESCC are unclear. Two cell lines (Clone 15 and 21) deficient for FGFRL1 expression were established from an ESCC cell line (KYSE520) using the CRISPR-Cas9 method. We evaluated the characteristics of these two cell lines in vitro and in vivo. In these FGFRL1-deficient KYSE520 cells, actin filaments around the nucleus were observed sparsely, whereas the filaments along the plasma membranes were observed as strong as those in the parent KYSE520 cells. Depleting FGFRL1 reduced cell motility in vitro and, decelerated tumor growth in a mouse xenograft model. Microarray analysis of mRNA expression revealed that FGFRL1-depletion resulted in reduced expression of MMP-1 and FGFBP1. In conclusions, FGFRL1-deficiency reduced both cell motility and tumor growth of KYSE520 cells. This is consistent with that high expression of FGFRL1 coincident with lymph node metastasis and poor prognosis. These results demonstrate that FGFRL1 expressed in ESCC cells has a role in aggressiveness of ESCCs.

Citation Format: Yoshinori Takei, Takafumi Matsumura, Kazuaki Watanabe, Hirokazu Nakamine, Tetsuo Sudo, Kazuharu Shimizu, Yutaka Shimada. FGFRL1 deficiency reduces motility and tumor growth of cells derived from esophageal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2513.
Abstract 252: Prostate carcinogenesis inhibition through AKT-PKCα signaling pathway by diosmetin
Authors: Pakradooni Rebecca , Khalifa Ahmad , Isali Ilaha , Shukla Sanjeev
Published: August 2018

Mammalian target of rapamycin (mTOR) integrates nutrient and mitogen signals to regulate cell growth (increased cell mass and cell size) and cell division. mTOR protein kinase have two different complexes; complex-I contains major component raptor, whereas complex II (insensitive to rapamycin), has major component rictor. Rictor plays a critical role in AKT (Ser-473 phosphorylation), which facilitates phosphorylation of AKT on Thr-308 by PDK1 for full activation. When we genetically disrupted Rictor, it decreased AKT phosphorylation (Ser-473) and impaired PKCα (Ser-657) expressions. Growth factors viz., IGF-1 and IL-6 are the major player in prostate cancers, these increases Rictor expression. We need to have an agent that can modulate kinase activity of AKT and PKCα to inhibit prostate cancer cell growth. Diosmetin (5, 7-Trihydroxy-4′-methoxyflavone) could be such an agent. Diosmetin, a natural flavone present in plant legumes, has anti-mutagenic and anti-allergic properties. Diosmetin inhibited IGF-1 and IL-6 induced rictor expression in LNCaP and PC-3 prostate cancer cells. Diosmetin dose response treatment to these cells inhibited cell growth and induced apoptosis with no significant growth inhibition in normal prostate epithelial cells (RWPE1). Diosmetin treatment to prostate cancer cells modulated cell survival machinery by down regulating key molecules viz., c-Myc, Survivin and XIAP (X-Linked Inhibitor of Apoptosis). Similarly diosmetin (20 and 50μg/animal/day) feeding to nude mice models, which were orthotopically implanted with luciferase tag PC-3 cells in ventral prostate represented significant decrease in tumor volume than control mice. Diosmetin fed tumor bearing mice reduced phosphorylation of Rictor, (Thr-1135), AKT (Ser-473) and PKCα (Ser-657) expressions to inhibit tumor growth significantly. Moreover, luciferase activity of implanted tumor decreased after diosmetin dose response feeding than control mice. Additionally bone radiographic images suggested that diosmetin treatment inhibited homing of PC-3 cells in the bone. These effects were associated with decrease in cyclin D1 expression and their activating partner, cyclin-dependent kinase (cdk) 2 and 4 with concomitant upregulation of p27/KIP1. Further diosmetin treatment induced apoptosis which was evident by increased expression of cleaved caspase-3. We are documenting these evidences for the first time in animal model system that diosmetin acts against potential molecular targets to alter cellular events to elicit anticancer effects in prostate cancer.

Citation Format: Rebecca Pakradooni, Ahmad Khalifa, Ilaha Isali, Sanjeev Shukla. Prostate carcinogenesis inhibition through AKT-PKCα signaling pathway by diosmetin [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 252.
Abstract 2538: The interaction of IGFBP3 with HIF2α in the invasion regulation of ovarian epithelial cell cancer
Authors: Shih Ho-Jun
Published: August 2018

Ovarian cancer is the most lethal gynecological cancer among women worldwide. The majority of malignant ovarian tumors are epithelial ovarian cancer and metastasis occurred in most patients at the time of diagnosis with worse outcome. New target therapy such as anti-angiogenesis therapy was proved to be effective to prolong progression free survival in epithelial ovarian cancer (EOC) patients. In our previous study, we have established an ovarian endometrioid carcinoma cell line OVTW59. By cDNA microarray analysis and further functional analysis, we have identified Insulin-like Growth Factor Binding Protein-3 (IGFBP-3), Calreticulin (CRT) and Thrombospondin-1(THBS1) correlated with tumor invasion in OVTW59. IGFBP3 and CRT was further found related with poor patient survival outcome. In this study, we focused on the mechanism of IGFBP3 in cancer invasion and metastasis. First we focused on the THBS1 related angiogenesis mechanism which was induced by hypoxia. Through this study, we identified HIF-2α, but not HIF-1α, playing as a significant role in EOC. By q-RT-PCR, Western blotting and immunohistochemical staining, HIF-2α showed opposite expression with IGFBP3, and high IGFBP3 was correlated with high THBS1 expression and low CRT expression. Using other cell lines, the IGFBP3/THBS1/HIF-2α/CRT expression patterns were similar in 293T, H1299 and A549. Furthermore, by Luciferase promoter assay, we verified that IGFBP3 could induce THBS1, inhibit HIF-2α expression, but does not regulate the expression of CRT. IGFBP3 is known as an invasion suppressor gene, THBS1 is an endogeneous inhibitor of angiogenesis, HIF-2α is a regulator of the hypoxia response and angiogenesis inducer, and CRT as a cell adhesion and invasion related protein. Overall, our study shows that IGFBP3 could induce tumor progression and metastasis through direct regulation of THBS1, and HIF-2α and indirect regulation of CRT in EOC.

Citation Format: Ho-Jun Shih. The interaction of IGFBP3 with HIF2α in the invasion regulation of ovarian epithelial cell cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2538.
Abstract 2554: To clone tumor-specific TCRs from a cell-based immunotherapy-benefit cervical cancer patient
Authors: Liang Xiaoling , Ou Shudan , Ma Minjun , Chen Xihe , Zhou Xiangjun , Han Yanyan
Published: August 2018

Adoptive transfer of tumor-specific T cell receptor (TCR)-engineered T cells, TCR-T therapy, has shown great efficacy in clinical trials against solid tumors. A cervical cancer patient with bone metastasis was treated with a cell-based immunotherapy, named Multiple-Antigens Stimulating Cellular Therapy (MASCT), which is a combination of multiple tumor antigen peptides loaded dendritic cells (DCs) and autologous T cells stimulated by these DCs. After repeated treatment, the patient showed partially response and remained stable disease for 20 months. Moreover, boosted specific immune responses were detected in patient's periphery blood by IFNγ-Elispot assay against various tumor antigens, such as CEA, RGS5 and HPV18/58. The clinical benefits of this patient indicated that tumor-specific T cells were expanded in vivo and played an important role to control tumor progression. These T cells may be the good sources to isolate tumor-specific TCRs for TCR-T therapies. Given that, patient's PBMCs were stimulated in vitro with selected tumor peptides respectively, such as CEA, RGS5 and HPV18 E7. Tumor-specific T cells were defined as IFNγ-secreting T cells upon peptide stimulation and were further enriched by using magnetic beads. Part of the enriched T cells were analyzed for TCR alpha/beta repertoire via NGS, the other part were sorted into single T cells and analyzed for TCR alpha/beta pairing. The TCR repertoire NGS data revealed that the tumor-specific T cells harbor several hundred unique TCR alpha/beta clonotypes. Some unique TCR alpha/beta clonotypes were considered as tumor-specific TCR candidates, since the copy numbersof these clonotypes were significantly and repeatedly increased after stimulation with CEA, RGS5 or HPV18 E7, and further increased after IFNγ enrichment. After TCR alpha/beta pairing confirmation by single T cell sequencing, alpha and beta chains of these TCRs were sent to synthesis for further investigation. In conclusion we have successfully identified specific TCR clonotypes targeting selected tumor antigens from a MASCT-benefited cervical cancer patient. After functional validations, including epitope specificity, HLA restriction as well as tumor recognition, these tumor-specific TCRs may be good candidates for developing safer and more effective TCR-T therapy targeting multiple tumor antigens.

Citation Format: Xiaoling Liang, Shudan Ou, Minjun Ma, Xihe Chen, Xiangjun Zhou, Yanyan Han. To clone tumor-specific TCRs from a cell-based immunotherapy-benefit cervical cancer patient [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2554.
Abstract 2575: NKX6.1 hypermethylation predicts the outcome of stage II colorectal cancer patients undergoing chemotherapy
Authors: Lin Ya-Wen , Chang Sou-Yi , Kuo Chih-Chi , Hsiao Cheng-Wen , Hsu Chih-Hsiung , Chou Yu-Ching , Shih Yu-Lueng
Published: August 2018

Background: Colorectal cancer (CRC) is a common malignancy worldwide. CRC patients in the same stage often present with dramatically different clinical scenarios. Thus, robust prognostic biomarkers are urgently needed to guide therapies and improve treatment outcomes. The NKX6.1 gene has been identified as a hypermethylation marker in cervical cancer and functions as a metastatic suppressor by regulating epithelial-mesenchymal transition (EMT). Here, we investigated whether hypermethylation of NKX6.1 might be a prognostic biomarker for CRC.

Results: In this study, we analyzed the methylation and expression of NKX6.1 in CRC tissues and CRC cell lines. We quantitatively examined the NKX6.1 methylation levels in 151 pairs of CRC tissues by using methylation-specific polymerase chain reaction (MS-PCR) analysis and found that NKX6.1 was hypermethylated in 35 of 151 CRC tissues (23.18%). NKX6.1 gene expression was inversely correlated with the DNA methylation level in CRC cell lines in vitro. Then, we analyzed the association of NKX6.1 methylation with clinical characteristics of these CRC patients. NKX6.1 methylation was positively associated with tumor size [odds ratio (OR): 2.84; 95% confidence interval (CI): 1.24 to 6.52]. These data demonstrated that patients with NKX6.1 methylation presented poorer 5-year overall survival (p=0.0167) and disease-free survival (p=0.0083) than patients without NKX6.1 methylation after receiving adjuvant chemotherapy. Most importantly, our data revealed that stage II CRC patients with NKX6.1 methylation had poorer 5-year disease-free survival (p=0.0322) than patients without NKX6.1 methylation after adjuvant chemotherapy.

Conclusions: Our results demonstrated that methylation of NKX6.1 is a novel prognostic biomarker in CRC and may be used as a predictor of the response to chemotherapy.

Citation Format: Ya-Wen Lin, Sou-Yi Chang, Chih-Chi Kuo, Cheng-Wen Hsiao, Chih-Hsiung Hsu, Yu-Ching Chou, Yu-Lueng Shih. NKX6.1 hypermethylation predicts the outcome of stage II colorectal cancer patients undergoing chemotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2575.
Abstract 2590: Race-specific differential expression of CCL25 and CCR9 in triple-negative breast cancer
Authors: King Jeronay , Kapur Neeraj , Mir Hina , Gales Dominique N. , Lillard James W. , Singh Shailesh
Published: August 2018

Breast cancer (BrCa) is the second leading cause of cancer-related deaths among women. Therapeutic intervention of the disease is impeded due to the heterogeneity contributed by inter- and intra-tumoral diversity, as well as difference among cancer-bearing individuals. Triple-negative breast cancer (TNBC) is a hormone-independent and highly aggressive subtype that disproportionally affects African-American (AA) women. However, the molecular basis of racial disparity in therapeutic response and clinical outcome in AA and Caucasian (CA) patients is still obscure. Our group was the first to show the involvement of CCR9 and its natural ligand, CCL25 in cancer progression and therapeutic response, including breast cancer. In this study, we have shown the association of CCR9 and CCL25 with TNBC. Both CCR9 and CCL25 are highly expressed in TNBC cells (MDA-MB-468 and MDA-MB-231) compared to other BrCa subtypes (MCF-7, MDA-MB-361, SKBr-3). Expression of this receptor-ligand pair was significantly elevated in TNBC cells derived from AA compared to those derived from CA patients. RNA-Seq based transcriptome data obtained from The Cancer Genome Atlas (TCGA) confirmed these results. Further, phospho-proteomic profiling of cancer signaling molecules showed race specific hyper-activation of GSK-3α, GSK-3β and NF-κB in AA derived cells. These molecules are known to provide the survival advantage to cancer cells. These findings, therefore, highlight significance of CCR9 and its association with the disparity in observed in TNBC. Hence, CCR9 targeted approach to address the disparity in the therapeutic outcome of TNBC seems to be promising.

Citation Format: Jeronay King, Neeraj Kapur, Hina Mir, Dominique N. Gales, James W. Lillard, Shailesh Singh. Race-specific differential expression of CCL25 and CCR9 in triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2590.
Abstract 2609: Overexpression of YAP1 of EGFR-mutant lung adenocarcinoma before tyrosine kinase inhibitor is associated with poor survival
Authors: Hong Soon Auck , Kang Jin-Hyoung , Hong Sook Hee
Published: August 2018

Purpose: EGFR tyrosine kinase inhibitor (EGFR TKI) was approved as a first line treatment for EGFR mutant lung adenocarcinoma (LADC) with advanced stage. YAP1 (Yes-associated protein 1) is a main effector of hippo pathway, related with adverse prognosis and EGFR TKI modulation of non-small cell lung cancer. This study aimed to clarify a prognostic role of YAP1 in EGFR mutant LADC and efficacy for EGFR TKI through the course of EGFR TKI.

Materials and Methods: 41 patients with paired lung cancer specimen before and after EGFR TKI were enrolled in this study. The expression of YAP1 protein was evaluated by immunohistochemistry.

Results: 15 cases (36.6%) with high YAP1 expression was found in pre-EGFR TKI LADC, while high YAP1 expression in 21 cases (52.5%) was detected after EGFR-TKI. The transitional level of YAP1 between pre- and post-EGFR TKI was significantly upregulated (P=0.002). High YAP1 before EGFR TKI was related with shorter OS (P=0.023) and PFS (P=0.041). In addition, high YAP1 before EGFR TKI in T790M mutant LADC was related with poor OS (P<0.001).

Conclusion: YAP1 burden before EGFR TKI was crucial role in prognosis of EGFR mutant LADC treated by EGFR TKI.

Citation Format: Soon Auck Hong, Jin-Hyoung Kang, Sook Hee Hong. Overexpression of YAP1 of EGFR-mutant lung adenocarcinoma before tyrosine kinase inhibitor is associated with poor survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2609.
Abstract 2614: Macrophage erythroblast attacher (MAEA) polymorphisms are associated with clinical outcome in TRIBE study mCRC patients treated with 5-fluorouracil/bevacizumab-based therapy
Authors: Soni Shivani , Miyamoto Yuji , Zhang Wu , Berger Martin D. , Cao Shu , Melendez Elizabeth , Puccini Alberto , Naseem Madiha , Tokunaga Ryuma , Battaglin Francesca , Mcskane Michelle , Cremolini Chiara , Falcone Alfredo , Loupakis Fotios , Lenz Heinz-Josef
Published: August 2018

Background: Macrophage erythroblast attacher (MAEA) also known as Emp (Erythroblast macrophage protein) plays indispensable role in erythroblast and macrophage development and targeted disruption of Emp is embryonically lethal (Soni et al., JBC 2006). Recently, down-regulation of Emp was implicated in abnormal cell motility and higher expression of mitogen-activated protein kinase 1 (MAPK 1) and thymoma viral proto-oncogene 1 (Akt 1) (Gulnaz et. al., BCMD 2016). Our preliminary studies, shows that higher expression of MAEA is associated with longer colorectal cancer (CRC) patient survival. We tested two MAEA polymorphisms (rs13149952 and rs1128427), whether these are correlated with clinical outcome in mCRC treated with 5-Fluorouracil / Bevacizumab based therapy. Methods: Genomic DNA was isolated from blood samples of two cohorts of mCRC patients. Cohort 1 included 227 patients treated with FOLFIRI/BEV (TRIBE trial Arm A, RAS wildtype/mutant=55/116, median age=60 years old, median follow-up period=49.3 months). Cohort 2 had 229 patients treated with FOLFOXIRI/BEV (TRIBE trial Arm B, RAS wildtype/mutant=60/115, median age=60 years old, median follow-up period=46.6 months). PCR-based direct Sanger sequencing was used to determine polymorphism. Results: Our results showed that MAEA rs1128427 in TRIBE arm A patients with RAS mutant tumors having any C genotype was significantly associated with longer overall survival as compared to TT genotype in univariate analysis {25.9 vs 18.8 months, HR(95% CI)=0.61(0.38,0.98), p=0.036}. In TRIBE arm B cohort, MAEA rs1128427 in patients with RAS mutant tumors carrying CC genotype showed significantly longer overall survival than patients with any T allele in both univariate and multivariable analyses {33.4 vs 25 months, uni: HR(95% CI)=0.57(0.34,0.95), p=0.028; multi: HR(95% CI)=0.56(0.32,0.96), p=0.034}. Conclusion: Our findings suggest for the first time, that polymorphisms in MAEA may predict the clinical outcome in Bevacizumab based therapies. Further prospective studies are warranted.

Citation Format: Shivani Soni, Yuji Miyamoto, Wu Zhang, Martin D. Berger, Shu Cao, Elizabeth Melendez, Alberto Puccini, Madiha Naseem, Ryuma Tokunaga, Francesca Battaglin, Michelle Mcskane, Chiara Cremolini, Alfredo Falcone, Fotios Loupakis, Heinz-Josef Lenz. Macrophage erythroblast attacher (MAEA) polymorphisms are associated with clinical outcome in TRIBE study mCRC patients treated with 5-fluorouracil/bevacizumab-based therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2614.
Abstract 2618: The thymidine kinase 1 (TK1) protein concentration in sera from Hodgkin lymphoma patients: A marker for cell death and disruption of malignant cells
Authors: Kumar Jagarlamudi Kiran , Eriksson Staffan , Ulfstedt Johan Mattsson , Venge Per , Molin Daniel
Published: August 2018

TK activity measurements have been used for many years to monitor cancer disease activity. However, what the blood concentrations of TK1 protein reflect is still uncertain. A TK1 immunoassay, the AroCell TK 210 ELISA, has been developed based on specific monoclonal antibodies against the C-terminal region of TK1. We utilized this assay to monitor the blood concentrations of TK1 in a cohort of patients with Hodgkin lymphoma (HL) before and after conventional treatment. TK1 concentrations in serum or plasma were measured using the AroCell TK 210 ELISA. For comparison, a large number of other biomarkers and cells were measured in blood such as C-reactive protein (CRP), Lactate dehydrogenase (LDH), hemoglobin, WBC, platelet counts, erythrocyte sedimentation rates (ESR), albumin, liver enzymes, creatinine. Fifty-eight patients with HL were included before start of treatment. Seventy three percent had the nodular sclerosis subtype and 52% stages I-II and 48% stages III-IV disease. Blood was sampled before treatment, during treatment and twice after completion of treatment. The upper normal TK1 concentration of healthy subjects (n=250) was 0.45 μg/L with a median of 0.25 μg/L (range 0.15-0.66 μg/L) as compared to HL, which was 0.26 μg/L (range 0.03-17 μg/L). The differences in ranges were significant (p<0.0001, F-statistics). Nodular sclerosis patients had higher TK1 levels as compared to the other subtypes (p=0.03). TK1 increased after treatment (p=0.001) in patients with normal concentrations from start i.e. <0.45 μg/L, whereas the concentrations decreased in 5/6 patients with TK1 >0.45 μg/L. TK1 concentrations before start of treatment was correlated to high CRP (p=0.01), low Hemoglobin (p=0.03), high WBC (p<0.001) and high platelet counts (p<0.01), but not to albumin or ESR or any other of the measured biomarkers. In a multivariate regression analysis, TK1 was correlated to LDH as the only independent variable (p<0.0001). In Hodgkin lymphoma patients, serum or plasma concentrations of TK1 may reflect malignant cell death and cell disruption rather than cell proliferation during treatment. Our results and hypothesis may help to interpret TK1 protein measurements as clinical tools in cancer management.

Citation Format: Jagarlamudi Kiran Kumar, Staffan Eriksson, Johan Mattsson Ulfstedt, Per Venge, Daniel Molin. The thymidine kinase 1 (TK1) protein concentration in sera from Hodgkin lymphoma patients: A marker for cell death and disruption of malignant cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2618.
Abstract 2626: Combination of chemotherapy and cytokine facilitates adoptive transferred effector T cells homing to the core of metastatic tumors
Authors: Hu Jiemiao , Zhao Qingnan , Xia Xueqing , Gorlick Richard , Li Shulin
Published: August 2018

Metastatic disease accounts for most cases of cancer mortality, due to its systemic nature and resistance to existing therapeutics for primary tumors. Immune therapy has become a viable alternative for treating drug-resistant tumors. In fact, increased immune density in metastasis lesions is associated with beneficial clinical outcomes. Notably, tumor infiltrating lymphocyte (TIL) infusions extended the overall survival of refractory melanoma patients. However, whether or not T cell therapy could be used to successfully limit the metastatic progression largely depends on the complexity of T cell penetration and tumor-immune cell interaction in distant sites. During the past years, our group has identified a simple combination of an immune stimulatory signal (interleukin-12) and chemotherapy (doxorubicin) that persistently induces NKG2D ligand on tumor cells in vivo across tumor types and developed a novel CD28 stimulation based approach for fast induction of NKG2D receptor on CD8+ T cells. We found in breast carcinoma model (4T-1) and pediatric osteosarcoma models (K7M3 and LM8), adoptive transfer of NKG2D+CD8+ T cells after the combination of IL-12 plus doxorubicin treatment dramatically reduced the numbers and sizes of metastatic nodules, leading to significantly prolonged survival time. Mechanically, the treatment of IL-12 plus doxorubicin widely upregulated the expression of NKG2D ligands exclusively on tumor cells in metastatic lesions, and allowed high density of NKG2D+CD8+ T cell accumulation at both the core and invasive margin areas of metastatic tumors. Furthermore, this combination therapeutic led to reduced regulatory T cell infiltration, resulting in increased CD8+ T cell to regulatory T cell ratios in metastatic tumors. Collectively, here we reported that a simple combination treatment of IL-12 plus doxorubicin with subsequent NKG2D+ CD8+T cell adoptive transfer may transform the metastatic tumor microenvironment to enable the influx of NKG2D+CD8+ T cells which effectively kill tumor cells via NKG2D-NKG2D ligand interaction. This promising therapeutic approach greatly improved the effectiveness and potential of adoptive T cell therapy in treating cancer patients with metastatic diseases.

Citation Format: Jiemiao Hu, Qingnan Zhao, Xueqing Xia, Richard Gorlick, Shulin Li. Combination of chemotherapy and cytokine facilitates adoptive transferred effector T cells homing to the core of metastatic tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2626.
Abstract 2650: Clinical implication of microsatellite instability in early gastric cancer
Authors: Choi Yoon Young , Kim Dong Gyu , Son Taeil , Kim Hyoung-Il , Cheong Jae-Ho , Hyung Woo Jin , Noh Sung Hoon
Published: August 2018

Purpose: The role of microsatellite instability is well established in advanced gastric cancer. However, its clinical implication has not been well addressed in early gastric cancer. The purpose of this study is to evaluate the clinical characteristics of microsatellite instability in early gastric cancer.

Methods:

Patients who underwent gastrectomy with curative intent in Severance hospital from January, 2005 to December, 2010 and diagnosed as early gastric cancer (pT1a/b) were enrolled in this study. Remnant gastric cancer and patients who received chemotherapy before operation were excluded. Microsatellite instability status was evaluated by two mononucleotide repeat markers (BAT25 and BAT26) and three dinucleotide repeat markers (D5S346, D2S123, and D17S250) and instability in two or more markers was defined as miscrosatellite instability high (MSI-H), otherwise classified as microsatellite stable (MSS).

Results: Of 1156 tumors that included the final analysis, 85 (7.4%) were MSI-H. MSI-H cancer was related to frequent female gender (54.1% vs. 32.8%, p<0.001), older age (63.4 ± 10.65 vs. 56.7 ± 11.50, p<0.001), lower body (81.2% vs. 59.7%, p=0.002), intestinal histology (63.5% vs. 48.2%, p=0.005), lympho-vascular invasion (25.9% vs. 13.3%, p=0.001), submucosal invasion (63.5% vs. 48.3%, p=0.007), and trend of lymph node metastasis (17.6% vs. 10.8%, p=0.056) compared to MSS type. Lymph node metastasis and lymph-vascular invasion was not differed by MSI status in mucosal gastric cancer (3.2% vs.

2.3%, p=0.755 and 3.2% vs. 2.3%, p=0.755, respectively). In sumbucosal gastric cancer, however, lympho-vascular invasion was frequently observed in MSI-H tumor (38.9% vs. 25.0%, p=0.027) but there was no difference in lymph node metastasis (25.9% vs. 19.9%, p=0.298). When we compared the disease free survival by MSI status, the prognosis of MSI-H tumor was similar compared to that of MSS tumor (log-rank test p=0.797, adjusted Hazard ratio of MSI-H by age, sex, pTstage and number of metastatic LNs: 0.932 [95% confidence interval: 0.423-2.054, p=0.861]).

Conclusions: MSI status could not be a useful biomarker in early gastric cancer to predict prognosis of it. However, frequent lympho-vascular invasion of MSI-H early gastric cancer would be a warning that careful patient's selection for endoscopic treatment or limited lymph node dissection for surgery is necessary.

Citation Format: Yoon Young Choi, Dong Gyu Kim, Taeil Son, Hyoung-Il Kim, Jae-Ho Cheong, Woo Jin Hyung, Sung Hoon Noh. Clinical implication of microsatellite instability in early gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2650.
Abstract 2663: CR42-24, a novel colchicine derivative, a therapeutic for bladder cancer
Authors: Bell Clayton , Potts Kyle , Pink Desmond , Lewis John , Tuszynski Jack
Published: August 2018

Colchicine is an anti-mitotic drug that targets unpolymerized tubulin, and inhibits microtubule polymerization. It is primarily used to treat gout but has been investigated in numerous clinical trials to treat conditions such as leukemia (ALL), prostate cancer, Behcet's disease, etc.. However, due to its narrow therapeutic window colchicine has had limited clinical translation. A possible approach to reduce general is to produce a derivative with an increased cancer specificity and selectivity by a narrower molecular target selection. Previously, our lab has designed and synthesized a novel colchicine derivative (CR42-24) with a more favorable pharmacological profile compared to colchicine. This was accomplished by designing a structure with an increased affinity for βIII tubulin. βIII tubulin is a β-tubulin isotype that is incorporated into tubulin dimers that make up microtubules. βIII is an excellent target for novel therapies as it has low expression in healthy tissue, is overexpressed in metastatic cancers, and is a clinical marker of poor prognosis. Using cell line screening we demonstrate that CR42-24 is highly toxic to a variety of cancer types with IC50 values at low nanomolar concentrations. More specifically CR42-24 is shown to be highly effective against bladder cancer. Current chemotherapy for bladder cancer is a combination of gemcitabine and cisplatin (Gem/Cis). Although marginally successful, many patients develop resistance to Gem/Cis leaving them with limited options for a second line therapy, thus development of alternative therapies is highly desired. Using in vitro assays we demonstrate that CR42-24 kills aggressive bladder cancer cell types, and is more effective than gemcitabine and cisplatin. Additionally, CR42-24 doubles survival rates of mice with bladder cancer xenografts. We also show that CR42-24 is highly synergistic with other chemotherapies, thereby increasing its therapeutic potential. Through our studies we have shown that CR42-24 is effective in treating aggressive bladder cancer and thus may serve as an alternative or second line therapy.

Citation Format: Clayton Bell, Kyle Potts, Desmond Pink, John Lewis, Jack Tuszynski. CR42-24, a novel colchicine derivative, a therapeutic for bladder cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2663.
Abstract 2676: Radiotherapy-assisted tumor selective metronomic oral chemotherapy
Authors: Cho Youngseok , Chung Seung Woo , Byun Youngro
Published: August 2018

Chemotherapy have commonly been used in maximum tolerated dose (MTD) to completely eradicate the cancer. However, such treatments often failed due to the complex and dynamic nature of cancer. Therefore, it has been suggested that cancer should be treated as a chronic disease, controlling its growth by providing continuous therapeutic pressure for long-term. Such an approach, however, requires a therapy that is non-toxic and orally available with sufficient potency. Herein, we propose a radiotherapy-assisted orally available metronomic apoptosis-targeted chemotherapy, which delivers doxorubicin continuously to the irradiated tumor with high selectivity while causing minimal toxicities to the normal tissues. DEVD-S- DOX/DCK complex is the anticancer prodrug for our strategy that could selectively release doxorubicin in the irradiated tumor tissue with sufficient oral bioavailability. The prodrug was completely inactive by itself, but displayed potent anticancer activity when coupled with radiotherapy. Consequently, the daily oral administration of DEVD-S-DOX/DCK in combination with the low-dose radiotherapy effectively suppressed the growth of tumor in vivo with no significant systemic toxicities despite that the accumulated dose of doxorubicin exceeded 150 mg/kg. Therefore, our novel therapy using DEVD-S-DOX/DCK complex is considered as an outstanding treatment option for treating cancer for long-term attributed to its oral availability and low-toxicity profile as well as the potent anticancer effect.

Citation Format: Youngseok Cho, Seung Woo Chung, Youngro Byun. Radiotherapy-assisted tumor selective metronomic oral chemotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2676.
Abstract 2679: Cyanobacterial trypsin-3 inhibitor inhibits prostate cancer cell invasion
Authors: Koistinen Hannu , Wahlsten Matti , Ahmed Muhammad N. , Sivonen Kaarina , Nees Matthias , Stenman Ulf-Håkan , Fewer David P.
Published: August 2018

Trypsin-3 is a highly active protease that has recently been identified as a potential therapeutic target for the reduction of tumor growth and metastasis in prostate, breast and pancreatic cancers. Current trypsin inhibitors typically inhibit a broad range of different trypsin-like enzymes. Thus, novel approaches to identify trypsin-3 selective inhibitors are needed. Cyanobacteria produce a vast diversity of natural products with complex chemical structures many of which are potent protease inhibitors.

We screened extracts of 140 cyanobacteria strains and discovered numerous strains showing selectivity towards trypsin-2 and -3 inhibition. We isolated a complex glycopeptide from one of these strains, which selectively inhibited human trypsin-2 and -3 with IC50 values of about 100 nM, while trypsin-1 was not inhibited. Importantly, we also found that this peptide inhibited invasion of aggressive and metastatic PC-3M prostate cancer cells though ECM preparation, while it did not affect the proliferation of the cells.

Our results suggest that microbial natural products may offer a viable alternative source of potent and selective trypsin-3 inhibitors. Such inhibitors may be suitable, after further development, for targeting the mechanisms associated with the invasion and metastatic dissemination of cancer cells, i.e., for treatment of aggressive cancers.

Citation Format: Hannu Koistinen, Matti Wahlsten, Muhammad N. Ahmed, Kaarina Sivonen, Matthias Nees, Ulf-Håkan Stenman, David P. Fewer. Cyanobacterial trypsin-3 inhibitor inhibits prostate cancer cell invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2679.
Abstract 2681: Antiproliferative effects of the naturally occurring solamargine and related glycoalkaloids against liver cancer cells
Authors: Fekry Mostafa I. , Ezzat Shahira M. , Salama Maha M. , Al-Abd Ahmed M.
Published: August 2018

Hepatocellular carcinoma (HCC) is progressively increasing tumor with lack of accurate prognosis and inadequate systemic treatment approaches. Solanum sp. (such as Solanum melongena) is a folk herb which is reported to possess anticancer properties. In a continuity for our interest in pursuing the anticancer activity of compounds isolated from the fruit peels of Solanum melongena, the HPLC profiling and ESI-MS assessment for the methanolic extract evidenced the presence of bioactive glycoalkaloids (solasonine, solasodine and solamargine). These glycoalkaloids were isolated, purified and proved to possess in vitro cytotoxicity against human liver cancer cell lines (Huh7 and HepG2). Herein, we investigated the potential mechanism of action of these compounds using DNA content flow-cytometry and apoptosis/necrosis differential anaylsis using annexin-V/FITC staining. Solasonine, solasodine and solamargine inducd significant antiproliferative effect against liver cancer cells (Huh7 and HepG2) which was attributed to cell cycle arrest at S-phase. Solamargine, solasodine and solasonine induced significant apoptosis in Huh7 cells. Only solamargine-induced cell cycle arrest, was reflected as apoptotic cell killing effect against HepG2 cells. In conclusion, glycoalkaloids derived from Solanum melongena and particularly, solamargine are promising antiproliferative agents with potential anticancer effects.

Citation Format: Mostafa I. Fekry, Shahira M. Ezzat, Maha M. Salama, Ahmed M. Al-Abd. Antiproliferative effects of the naturally occurring solamargine and related glycoalkaloids against liver cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2681.
Abstract 2687: Cypripedin inhibits non-small cell lung cancer migration through suppression of epithelial-to-mesenchymal transition
Authors: Treesuwan Surassawadee , Pongrakhananon Varisa
Published: August 2018

Lung cancer is one of the most common cause of cancer-related death worldwide because of high metastasis rate. Cancer migration is a prerequisite event among multi-complex processes of metastasis, being one of the interested target for cancer therapy. Accumulative evidence emphasizes that epithelial-to-mesenchymal transition (EMT) plays an important behavior exerting cell motility and survival, and relevant to cancer metastasize. Even the therapeutic approaches have been developed, lung cancer metastasize remains challenge for drug discovery and development. Recent studies highlight the potent anticancer properties of the compounds derived from Orchid. However, the underlying mechanism was unrevealed. This study demonstrated that cypripedin, a phenanthrenequinone extracted from Dendrobium densiflorum, possess an ability to inhibit in vitro cell migration in non-small cell lung cancer H460 and H23 cells. The number of motile cells, examined by transwell migration and wound healing assay, were gradually reduced in a dose-dependent concentration of cypripedin, along with the reduction of colony number and size formed under an anchorage-independent growth. Mechanistic investigation shows that the mesenchymal markers including N-Cadherin, Vimentin and Slug were significantly down-regulated following cypripedin treatment. Our finding suggests that cypripedin potentiates anti-migrative activity in non-small cell lung cancer through the attenuation of EMT process, which support further extensive studies against lung cancer.

Citation Format: Surassawadee Treesuwan, Varisa Pongrakhananon. Cypripedin inhibits non-small cell lung cancer migration through suppression of epithelial-to-mesenchymal transition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2687.
Abstract 2701: Proteogenomic analysis of endoscopy biopsy identifies alterations in apoptotic pathways in trastuzumab-resistant gastric cancers
Authors: Kang Jeong Won , Choi Il Ju , Kim Hark Kyun
Published: August 2018

Although the combination of trastuzumab with cisplatin and fluoropyrimidine is standard therapy for metastatic HER2-positive gastric cancers, predictive biomarkers for this therapy need to be further refined. Given the multifactorial nature of clinical trastuzumab resistance, we performed nano-liquid chromatography (LC)-tandem mass spectrometry (MS/MS) proteomic analyses on 13 pairs of frozen HER2-positive tumors and adjacent normal tissue counterparts collected from patients who were treated with trastuzumab in combination with cisplatin and fluoropyrimidine (TCF). We quantified >6,000 protein groups per tumor on the average, and this number was almost comparable to those obtained by NCI's Clinical Proteomic Tumor Analysis Consortium projects, which characterized resected tumors using similar nano-LC-MS/MS technologies. Nano-LC-MS/MS data for differentially expressed proteins were successfully validated by our immunostaining analyses. When we correlated proteomic data with clinical outcome, underexpression of apoptosis pathway proteins was the prominent proteomic signature in TCF-resistant tumors, as compared with TCF-sensitive tumors. Proapoptotic proteins in FAS signaling and mitochondrial apoptosis pathways were less abundant in TCF-resistant tumors than in sensitive tumors. When iTRAQ reporter ion ratios were compared between pretreatment and acquired-resistant tumors from clinical responders, apoptosis pathways were significantly enriched in proteins that were overexpressed in the acquired-resistant tumors. Assuming that the pretreatment tumors of clinical responders were mostly composed of chemosensitive clones, these antiapoptotic proteins may confer survival advantage on resistant clones, resulting in the repopulation of resistant clones overexpressing these antiapoptotic proteins. For the first time to our knowledge, therefore, our proteogenomic study of frozen endoscopic biopsy samples revealed that alterations in apoptotic pathway proteins were most significantly associated with clinical trastuzumab resistance. Our study is also the first to demonstrate that nano-LC-MS/MS technology can be applied to small biopsy tissue samples for precision medicine research.

Citation Format: Jeong Won Kang, Il Ju Choi, Hark Kyun Kim. Proteogenomic analysis of endoscopy biopsy identifies alterations in apoptotic pathways in trastuzumab-resistant gastric cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2701.
Abstract 2722: Investigation of the expression profile and functional role of PD-1, TIM-3 and LAG-3 in human tumors
Authors: Ghosh Srimoyee , Travers Jon , McEachern Kristen , Kumar Sujatha , Ramaswamy Sridhar , Jenkins David
Published: August 2018

While immunotherapies directed against PD-1 and PD-L1 have proven effective across multiple indications, there is still a large unmet medical need for therapies for patients who do not respond or who develop acquired resistance during the course of treatment. There are several emerging hypotheses to explain the lack of response, one of which is the upregulated expression of additional checkpoint receptors, such as TIM-3 and LAG-3, that limit the ability of PD-1 pathway blockade to re-establish effective anti-tumor immunity.

To investigate the potential role of TIM-3 and LAG-3 relative to PD-1, we set out to evaluate the expression and function of these receptors in several systems. Firstly, we used a flow cytometry approach to enumerate immune cell populations in a panel of human tumor samples, including non-small cell lung cancer (NSCLC). In these samples, PD-1, TIM-3 and LAG-3 expression was observed in both T-cell and non-T-cell populations. PD-1 was found to be co-expressed with TIM-3 and LAG-3 in CD8+ T-cells and we also found expression of TIM-3 and LAG-3 in other cell types, including regulatory T-cells and myeloid cells.

Building on the expression data, we explored the functional effects of targeting these receptors, evaluating the effect of single agent and combination effects of anti-PD-1, anti-TIM-3 and anti-LAG-3 antibodies in mouse models. In these studies, we found not only that dual-blockade of PD-1 with either anti-TIM-3 or anti-LAG-3 resulted in improvements in efficacy over monotherapy, but also that the triple combination of all 3 inhibitors was associated with further anti-tumor activity. In a humanized mouse model of NSCLC, the triple combination was associated with pharmacodynamic effects not only in T-cells, but also reductions in Tregs and macrophages.

Taken together, these studies provide further evidence to suggest that in addition to PD-1, LAG-3 and TIM-3 are important emerging immunotherapy targets and provide rationale for not only doublet but also triplet combinations as an approach to cancer therapy.

Citation Format: Srimoyee Ghosh, Jon Travers, Kristen McEachern, Sujatha Kumar, Sridhar Ramaswamy, David Jenkins. Investigation of the expression profile and functional role of PD-1, TIM-3 and LAG-3 in human tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2722.
Abstract 2729: Quantitative cell-based bioassays to advance individual or combination immune checkpoint immunotherapy
Authors: Grailer Jamison , Gilden Julia , Stecha Pete , Garvin Denise , Hartnett Jim , Fan Frank , Cong Mei , Cheng Zhi-Jie Jey
Published: August 2018

Immune checkpoint receptors play a critical role in maintaining immune homeostasis and are genetically and functionally associated with autoimmune disease, cancer and persistent viral infections. Blockade of immune checkpoints has emerged as a promising new approach to enhance anti-tumor immune responses. After initial successes with PD-1 and CTLA-4-targeted therapies, immunotherapy research has broadened to include additional immune checkpoint receptors targeted individually or in combination. A significant challenge in the development of biologics that target immune checkpoints is access to quantitative and reproducible functional bioassays. Existing methods rely on primary cells and measurement of complex functional endpoints. As an alternative to primary cell assays, we have developed a panel of cell-based reporter bioassays that can quantitatively determine the potencies of biologics targeting the immune checkpoints PD-1, CTLA-4, LAG-3, TIM-3, TIGIT, and BTLA, or bispecific antibody or antibody cocktails targeting two immune checkpoint receptors such as PD-1+TIGIT, PD-1+LAG-3, and PD-1+CTLA-4. These assays consist of engineered T effector cells that express luciferase reporters driven by specific response elements integrating signaling from the T cell receptor and immune checkpoint target(s), and engineered artificial antigen presenting cells. These mechanism of action-based bioassays demonstrate high specificity, sensitivity and reproducibility. Therefore they can serve as valuable tools for immunotherapy drug development.

Citation Format: Jamison Grailer, Julia Gilden, Pete Stecha, Denise Garvin, Jim Hartnett, Frank Fan, Mei Cong, Zhi-Jie Jey Cheng. Quantitative cell-based bioassays to advance individual or combination immune checkpoint immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2729.