bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5892: Whole-exome and RNA sequencing reveals chromosomal rearrangements associated with the recurrence of pulmonary carcinoid
Authors: Miyanaga Akihiko , Masuda Mari , Motoi Noriko , Tsuta Koji , Nakamura Yuka , Nishijima Nobuhiko , Watanabe Shun-ichi , Asamura Hisao , Tsuchida Akihiko , Seike Masahiro , Gemma Akihiko , Yamada Tesshi
Published: September 2020

Background: The majority of pulmonary carcinoid (PC) tumors can be cured by surgical resection alone, but a significant proportion of patients experience recurrence. As PC is insensitive to conventional chemotherapy and a significant proportion of patients develop metastases, further clarification of the molecular mechanisms of metastasis is needed in order to develop targeted therapeutics.

Material and Method: We performed comprehensive whole-exome sequencing (WES) of primary tumors and corresponding normal lung tissues from 14 PC patients (including 4 patients who developed postsurgical distant metastasis) and RNA sequencing of primary tumors from 6 PC patients (including 4 patients who developed postsurgical distant metastasis). Exon array-based gene expression analysis was performed in 25 cases of PC.

Results: We identified a total of 139 alterations in 136 genes. MUC6 and SPTA1 were recurrently mutated at a frequency of 21% (3/14) and 14% (2/14), respectively. Mucin protein family genes including MUC2, MUC4 and MUC6 were mutated in a mutually exclusive manner in 36% (5/14). Pathway analysis of the mutated genes revealed enrichment of genes involved in mitogen-activated protein kinase (MAPK) signaling, regulation of the actin cytoskeleton and focal adhesion, and transforming growth factor (TGF)-β signaling. RNA sequencing revealed a total of 8 novel fusion transcripts including one derived from a chromosomal translocation between the TRIB2 and PRKCE genes. All of the 8 fusion genes were detected in primary PCs that had developed metastasis after surgical resection. We identified 14 genes up-regulated in 5 PCs that had relapsed after surgical resection.

Conclusions: In this study we identified novel somatic mutations and chromosomal rearrangements in PC by examining clinically aggressive cases that had developed postsurgical metastasis. It will be essential to validate the clinical significance of these genetic changes in a larger independent patient cohort.

Citation Format: Akihiko Miyanaga, Mari Masuda, Noriko Motoi, Koji Tsuta, Yuka Nakamura, Nobuhiko Nishijima, Shun-ichi Watanabe, Hisao Asamura, Akihiko Tsuchida, Masahiro Seike, Akihiko Gemma, Tesshi Yamada. Whole-exome and RNA sequencing reveals chromosomal rearrangements associated with the recurrence of pulmonary carcinoid [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 5892.
Abstract 590: Efficacy evaluation of bispecific antibodies in humanized mice after B cell depletion
Authors: Gan Tian , Li Dirui , Yang Hao , Ng Gary , Li Yanan , Lin Qingcong
Published: September 2020

The development of bifunctional/multifunctional therapeutics have gained tremendous traction in recent years. Preclinical efficacy evaluation of bi-functional antibody drugs faces two main problems: first is the often limited cross-reaction between the anti-human antibody and murine target in animals; second is the reduced efficacy as a result of neutralization of antibodies by anti-drug antibodies (ADA) in animals due to the high immunogenicity of some bi-specific antibodies.

To eliminate the ADA production against bi-specific therapeutic agents, Biocytogen developed a B cell depletion method based on humanized mouse syngeneic xenograft tumor model for preclinical efficacy evaluation of bispecific antibodies. To a syngeneic xenograft tumor model (PD-L1 humanized mice inoculated subcutaneously with MC38-hPD-L1 cells), CD20 antibody was injected to deplete B cells in the mice for the inhibition anti-antibody production. A model bi-specific agent against PD-L1 and TGF-β was sequentially injected to the mice. The tumor inhibitory effect of the bi-specific was assessed by tumor growth inhibition rate (TGI) calculated from measured tumor volume.

Blood samples were drawn from mice for B cell analysis. The clearance of B cells in the blood flow and the serum concentration (including ADA) in mice were analyzed by flow cytometry and ELISA, respectively. The results showed that B cells were effectively eliminated from the murine blood after injection of CD20 antibody. Despite of the absence of B cells, the tumor growth was inhibited upon the treatment with M7824 analog, compared with the control group. Corresponding to the efficacy results, the mice in B cells-depleted group maintained a normal drug concentration in the blood while reducing the production of anti-drug antibodies. In summary, the B cell depletion mice have been validated as a promising model for the efficacy evaluation of complex structure antibody (bi or multi-functional antibody).

Citation Format: Tian Gan, Dirui Li, Hao Yang, Gary Ng, Yanan Li, Qingcong Lin. Efficacy evaluation of bispecific antibodies in humanized mice after B cell depletion [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 590.
Abstract 5912: YTHDF1 amplification associates with clinicopathological featuresand predicts worse survival in breast cancer
Authors: Li Cheukfai , Zhang Guochun , Wang Yulei , Chen Bo , Li Kai , Cao Li , Ren Chongyang , Wen Lingzhu , Jia Minghan , Mok Hsiaopei , Lai Jianguo , Xiao Weikai , Li Xuerui , Liao Ning
Published: September 2020

N6-methyladenosine (m6A) is a common RNA modification on eukaryotic mRNA and several m6a regulatory proteins, which plays a crucial part in breast cancer. But the copy number variations (CNVs) for m6a regulatory proteins and their role in pathogenesis and progression in breast cancer is still unclear. By analyzing the Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) and the cancer genome atlas (TCGA database), we screened CNVs of ten m6A regulatory genes, including YTHDF3, YTHDF1, FTO, YTHDC1, ALKBH5, METTL3, WTAP, METTL14, YTHDF2, YTHDC2. Among them, YTHDF3 and YTHDF1 amplification had higher alteration rates among ten m6A regulatory genes. YTHDF1 and YTHDF3 amplification resulted in higher mRNA expression (P<0.001). We found that YTHDF1 and YTHDF3 amplification presented a high correlation with worse cinlicopathological characteristics and overall survival (OS) in breast cancer patients. COX regression analysis showed YTHDF1 amplification was an independent risk factor for 10-year OS in breast cancer (HR=1.549, 95% CI: 1.408-1.705, P<0.001, Table). Moreover, GSEA analysis revealed the downstream target of YTHDF1 may be related to MYC signaling regulation and T cell differentiation. In conclusion, we illustrated genetic alteration of m6A regulatory genes in breast cancer patients and found significant relationship between YTHDF1 amplification and worse clinical characteristics, indicating it a potential target for breast cancer treatment in epigenetic modification.

Univariate and multivariate Cox proportional hazard analysis for 10-year overall survivalUnivariateUnivariateHR95% CIP valueHR95% CIP valueNode status1.7181.566-1.885<0.0011.5491.408-1.705<0.001Tumor Size1.0161.013-1.019<0.001ER status0.6200.530-0.725<0.0010.5260.440-0.629<0.001PR status0.6120.531-0.707<0.001HER2 status1.7471.445-2.112<0.0011.5171.234-1.866<0.001Age1.0241.018-1.030<0.0011.0331.026-1.039<0.001YTHDF11.8111.419-2.313<0.0011.5491.408-1.705<0.001

Citation Format: Cheukfai Li, Guochun Zhang, Yulei Wang, Bo Chen, Kai Li, Li Cao, Chongyang Ren, Lingzhu Wen, Minghan Jia, Hsiaopei Mok, Jianguo Lai, Weikai Xiao, Xuerui Li, Ning Liao. YTHDF1 amplification associates with clinicopathological featuresand predicts worse survival in 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 5912.
Abstract 5914: URI participates in cisplatin-induced DNA damage repair of gastric cancer cells via the ATM/CHK2 pathway
Authors: Bian Huiqin , Lu Yaojuan , Gu Yu , Zhang Xiaojing , Sun Lichun , Gu Junxia , Zheng Qiping
Published: September 2020

Gastric cancer is the most common gastrointestinal malignant tumor and also the second leading cause of cancer-related mortality worldwide. Given its high rate of non- symptoms at early stage, gastric cancer is usually diagnosed at middle or advanced stage with metastasis. Therefore, chemotherapy, including Cisplatin, becomes a main treatment option, in addition to surgery. However, it is not uncommon for patients to develop cisplatin resistance which limits its clinical applications. URI, a prefoldin family member and also an oncoprotein, has been shown to promote multiple cancer development, including gastric cancer. The aim of this study was to investigate how URI may participate in the DNA damage repair of gastric cancer cells induced by Cisplatin and its possible mechanism via the ATM/CHK2 pathway. URI knockdown cell lines from gastric cancer cells MGC-803 and SGC-7901 were established and treated with different concentrations of Cisplatin. Cell viability, proliferation and apoptosis were analyzed by CCK-8, EDU assay, and flow cytometry respectively. DNA injury marker H2AX were examined by qRT-PCR and Western blot analysis. Comet assay was used to detect the DNA damage. The results showed that URI knockdown reduces cell viability and proliferation, as well as the Cisplatin-induced apoptosis of gastric cancer cells. Cisplatin dose responsively induced DNA damage in gastric cancer cells and URI knockdown enhanced this damage as indicated by comet assay and increased γH2AX expression. We have also detected increased levels of P-ATM and P-CHK2 in gastric cancer cells treated with Cisplatin, while URI knockdown decreased the P-ATM and P-CHK2 expression after the cells were treated with cisplatin for 12 hours. Taken together, our results support that URI reduces DNA damage and promotes DNA repair of gastric cancer cells induced by cisplatin and therefore, enhances its resistance possibly via the ATM/CHK2 pathway

Citation Format: Huiqin Bian, Yaojuan Lu, Yu Gu, Xiaojing Zhang, Lichun Sun, Junxia Gu, Qiping Zheng. URI participates in cisplatin-induced DNA damage repair of gastric cancer cells via the ATM/CHK2 pathway [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 5914.
Abstract 5924: A clock gene of TIM promotes colorectal cancer tumorigenesis through binding with myosin-9
Authors: Yuan Xiaoqin , Wang Yi , Wu Lei
Published: September 2020

Colorectal cancer (CRC) is a common tumor that is characterized by its high mortality rate. However, the underlying molecular mechanisms that drive CRC tumorigenesis are still not clear. Clock genes have been recognized as playing key roles in tumor development. In our present study, we found the expression of TIM (encoding protein of Timeless), one of the clock genes was up-regulated in CRC tissues comparing to the corresponding normal tissues by the results of IHC and its expression was closely associated with tumor clinical stage and poor overall survival of CRC patients through analysis of clinical CRC patients. Functional studies demonstrated that TIM promoted CRC cell proliferation, migration and invasion in vitro and in vivo. Mechanistic investigations showed that TIM plays a key role in EMT. Immunoprecipitation followed by mass spectrometry revealed that Timeless bound with Myosin-9. Furthermore, we also found TIM regulated the expression level of Myosin-9. And Myosin-9 knockdown retarded the migration and invasion of TIM-overexpressed CRC cells. Furthermore, we found the expression of TIM was regulated by histone acetylation at its promoter area. Our results show that TIM could regulate tumorigenesis of CRC by binding and regulating Myosin-9. Our results also implicate TIM as a potential prognostic biomarker and therapeutic target for CRC.

Citation Format: Xiaoqin Yuan, Yi Wang, Lei Wu. A clock gene of TIM promotes colorectal cancer tumorigenesis through binding with myosin-9 [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 5924.
Abstract 5943: The role of IL-6 family cytokines in gastrointestinal cancer
Authors: Taniguchi Koji , Kawazoe Tetsuro
Published: September 2020

The important effector of the Hippo pathway, YAP, is strongly activated in the majority of gastrointestinal cancer. However, the mechanism by which YAP is activated in gastrointestinal cancer remains unknown. Previously, we found that after intestinal injury, IL-6 family cytokines activate the Src family kinases (SFK)-YAP pathway, which enhances intestinal regeneration. Here, we found that in addition to β-catenin, loss of tumor suppressor adenomatous polyposis coli (APC) activates SFKs, YAP and STAT3, which are simultaneously activated in 60-70% of human colorectal cancer specimens. We also found that up-regulation of IL-6 signal transducer (IL-6ST/gp130), which is induced by YAP activation, causes the activation of SFKs, YAP and STAT3. We also found that leukemia inhibitory factor (LIF), one of the IL-6 family cytokines, plays an important role in human esophageal squamous cell carcinoma by activating the SFK-YAP pathway. The combination treatment of SFK and JAK inhibitors results in suppression of gastrointestinal tumors. These results suggest that these signaling pathways might be new attractive therapeutic targets in human gastrointestinal cancer.

Citation Format: Koji Taniguchi, Tetsuro Kawazoe. The role of IL-6 family cytokines in gastrointestinal 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 5943.
Abstract 5957: Roles for the cancer stem cell marker Aldh1A1 in Merlin null nervous system tumors
Authors: Hillson Lily , Laraba Liyam , Shivane Aditya G. , Edwards Philip , Bantukallu Ganesha , Simeonov Anton , Martinez Natalia J. , Yang Shyh-Ming , Parkinson David B.
Published: September 2020

Our lab is interested in the development and growth of schwannoma and meningioma tumors of the nervous system. Approximately 70% of these tumors arise due to loss of the tumor suppressor Merlin. In a mouse model of schwannoma (P0-CRE NF2fl/fl), we observed increased mRNA levels of the cancer stem cell marker Aldehyde dehydrogenase 1A1 (Aldh1A1) in Merlin-null Schwann cells. Immunostaining showed elevated levels of Aldh1A1 protein in this mouse model and in another mouse model (Postn-CRE NF2fl/fl), in which schwannoma tumors arise spontaneously within the vestibulocochlear nerve and dorsal root ganglia of the nervous system. Similarly, staining of human schwannoma samples also showed a strong elevation of Aldh1A1 protein in all tumor cells. Parallel analysis of meningioma tumor samples, in which Merlin loss also occurs, also showed a strong increase of Aldh1A1 protein levels. Western blotting enabled identification of the mechanism by which Merlin loss regulates Aldh1A1. Merlin/TAZ double knockout, but not Merlin/YAP double knockout, sciatic nerves show reduced Aldh1A1 expression. This implicates Hippo signalling through the TAZ effector as the mechanism of Aldh1A1 upregulation both in vitro and in vivo in both tumor types. Chemical inhibition with Aldh1a1-specific inhibitor (NCT-505) or shRNA knockdown of Aldh1A1 in Merlin-null schwannoma and meningioma cells in culture attenuated tumor cell growth, proving Aldh1A1 has a role in tumor cell proliferation; studies using Aldh1A1 null mice to test this in vivo are ongoing. A major target of Aldh1A1 signalling is the regulation of retinoic acid (RA) signalling. We are examining expression and activation of components of RA signalling in both schwannoma and meningioma tumor cells to identify whether this may be of therapeutic use in the future for these two tumor types. In conclusion, the TAZ-dependent upregulation of Aldh1A1 in Merlin-null schwannoma and meningioma tumor cells has a functional role in driving cell proliferation in these two clinically important tumor types. Further work will seek to identify the mechanism by which Aldh1A1 activity contributes to tumor growth.

Citation Format: Lily Hillson, Liyam Laraba, Aditya G. Shivane, Philip Edwards, Ganesha Bantukallu, Anton Simeonov, Natalia J. Martinez, Shyh-Ming Yang, David B. Parkinson. Roles for the cancer stem cell marker Aldh1A1 in Merlin null nervous system tumors [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 5957.
Abstract 598: Integrated computational and experimental design of a monobody targeting PDL1
Authors: Harrison Reed E. , Man Chi-Wei , Wang Yingxiao
Published: September 2020

Monobodies (Mb) are small, synthetic antibodies based on the human fibronectin type III domain that exhibit high thermodynamic stability without disulfide bonds. To explore suitability of these domains as cancer-targeting molecules, we apply computational and experimental methods to develop a Mb that targets programmed-death-ligand 1 (PDL1). As some tumors overexpress PDL1 to inhibit T-cell responses, a PDL1-binding Mb could be used to promote successful immunotherapy. More broadly, computational methods applied to developing a PDL1-binding Mb can also be applied to developing new Mbs that target cancer antigens. Starting from the G9 Mb (PDB: 1ttg), we grafted the CDR3 loop of a known PDL1-binder (PDB: 5jds) into the FG loop of the Mb. This design was displayed on the surface of yeast and shown to bind PDL1 better than the G9 Mb, though not as well as a high affinity variant of PD1, the cognate receptor of PDL1. Using molecular dynamics simulations, we evaluated structural features that are unlikely to contribute to PDL1-binding. Using this approach, we selected amino acid positions for site-saturated mutagenesis and constructed a yeast library displaying Mb isoforms. Through iterative rounds of selection, we identified a Mb isoform with further improved PDL1-binding.

Citation Format: Reed E. Harrison, Chi-Wei Man, Yingxiao Wang. Integrated computational and experimental design of a monobody targeting PDL1 [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 598.
Abstract 5986: Inverse relationship between Giant Axonal Neuropathy gene expression and EMT in human tumors
Authors: Veena Mysore S. , Shin Daniel Sanghoon , Jeong Chan , Chatoff Jenna R. , Venkatesan Natarajan , Basak Saroj K. , Gray David , Wilczynski Sharon P. , Wilczynski Sharon P. , Gray Steven J. , Kohn Donald B. , Wang Marilene B. , Srivatsan Eri S.
Published: September 2020

Gigaxonin, the protein product of the Giant axonal neuropathy (GAN) gene, is a E3 ubiquitin ligase involved in neural cell and fibroblast intermediate filament processing. We have previously shown that Gigaxonin interacts with p16 in cisplatin-mediated ubiquitination of NF-κB. We analyzed cervical and head and neck cancer cell lines and primary tumors and have found exon 8 c.1293C>T single nucleotide polymorphism (SNP) to occur in both HPV positive and negative tumor cells lines and primary tumors. The prevalence of this SNP in the tumor samples was double that of the normal population (47.25% vs. 22%) and there was no relationship to HPV status. Cell lines containing the SNP showed higher expression of gigaxonin, and an inverse relationship to in vitro tumor cell line growth and cisplatin sensitivity. There was a direct relationship between the expression of gigaxonin and e-cadherin and inverse relationship to the expression of twist, snail and n-cadherin. CRISPR-Cas9 mediated conversion of T to C allele in gigaxonin overexpressing cervical cancer cell line ME 180 resulted in decreased gigaxonin expression accompanied by increased expression of twist, and snail and increased in vitro cell growth. Re-expression of gigaxonin in the CRISPR-Cas9 clone with a plasmid gigaxonin cDNA construct resulted in decreased expression of twist pointing to gigaxonin mediated downregulation of the EMT marker. We therefore hypothesize that GAN, associated with the neurodegenerative disease, is also a human tumor cell growth suppressor gene.

Citation Format: Mysore S. Veena, Daniel Sanghoon Shin, Chan Jeong, Jenna R. Chatoff, Natarajan Venkatesan, Saroj K. Basak, David Gray, Sharon P. Wilczynski, Sharon P. Wilczynski, Steven J. Gray, Donald B. Kohn, Marilene B. Wang, Eri S. Srivatsan. Inverse relationship between Giant Axonal Neuropathy gene expression and EMT in human tumors [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 5986.
Abstract 5999: The Warburg Effect suggests a palliative care approach to cancer therapy
Authors: Meade-Tollin Linda C. , Tsang Tom C.
Published: September 2020

Introduction: Otto Warburg discovered in 1924 that cancer cells needed a lot more glucose than normal cells. Today, the “Warburg Effect” is not only recognized as a “hallmark of cancer” but also served as the scientific basis for cancer detection using fluorodeoxyglucose (FDG) Positron emission tomography (PET-scans). Warburg suggested in 1927 that treating cancer patients with insulin can deprive glucose to the cancer cells by lowering the blood glucose levels and may inhibit tumor growth. Although at least six peer-reviewed publications so far have provided support for the feasibility of this approach, no clinical trial has ever been done. We have previously presented an analysis of the data from hospitals in China suggesting that insulin can be safely administered for patients receiving a small dosage of standard chemotherapy drugs as palliative treatments for late-stage cancer patients. Here, we report additional data from more patients and medical facilities.

Experimental procedures: Patients diagnosed with multiple types of late-stage solid tumors were treated with regular insulin (Humulin R) administered intravenously, and small doses of standard chemotherapy drugs (approximately 10% of the standard dosages). Blood glucose levels were monitored throughout the course of treatment. Hypoglycemia was reversed by ingestion of food and drink or injection of glucose solution. Depending on how well the patients tolerated the treatments, 5-30 units of insulin were used. No permanent side effects were observed or reported.

Summary of data: Over 400 treatments have been performed on over 30 patients ranging from 30 to 76 years old, and diagnosed with 14 types of cancer. The average age of the patients was 56.2 years old. Symptoms of hypoglycemia were observed in all 30+ patients, and readily reversed by intravenous infusions of glucose or ingestion of food and drink. The insulin treatments were well tolerated and no longer -term side effects were observed.

Conclusions: Our analysis supports the feasibility of using regular insulin as a reproducibly safe method to achieve glucose deprivation in patients with advanced stages of multiple types of cancer receiving palliative and hospice care. These and additional future studies could form the basis for eventually conducting a clinical trial testing whether regular insulin, which is inexpensive and available over the counter, can be an alternative therapeutic option for palliative and hospice care in some patients.

Citation Format: Linda C. Meade-Tollin, Tom C. Tsang. The Warburg Effect suggests a palliative care approach to cancer 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 5999.
Abstract 6000: eIF3A regulate the Warburg effect via ENO1 in cervical cancer cells
Authors: Xu Pengfei , Li Dake
Published: September 2020

Eukaryotic translation initiation factor 3A (eIF3A) is one of the core subunits of the translation initiation complex eIF3. eIF3A is critical for ribosomal subunit joining and recruiting mRNA to the ribosome, as well as for the regulation of various gene products. However, the function of eif3A have not been reported in cervical cancer (CC). Here we reported that Eif3A protein was overexpressed in CC tissues and the higher expression of Eif3A predicted a worse prognosis. Further experiments showed that eIF3A knockdown inhibited CC cells' proliferation and migration and promoted apoptosis in vitro and in vivo. eIF3A silencing accelerated acidification of the culture medium of CC cells. Seahorse results showed that the decreased expression of eIF3A significantly reduced glycolysis flux in CC cells. Mass spectrometry suggested that eIF3A knockdown could significantly repressed the expression of ENO1, which is a famous glycolytic protein. Western blot assay indicated that eIF3A silencing significant decreased the expression of ENO1. Therefore, our results indicated that eIF3A could enhance the warburg effect in cervical cancer by regulating ENO1.

Citation Format: Pengfei Xu, Dake Li. eIF3A regulate the Warburg effect via ENO1 in cervical 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 6000.
Abstract 6006: Total body irradiation and graft-versus-host disease: A meta-analysis
Authors: Poonsombudlert Kittika , Prueksapraopong Chattip , Kewcharoen Jakrin , Limpruttidham Nath
Published: September 2020

Objective: Total Body Irradiation (TBI) has been one of the standard conditioning regimens since the early development of allogeneic hematopoietic stem cell transplant (HSCT). Some studies have suggested a correlation between the use of TBI and the development of acute and chronic graft-versus-host disease (aGVHD and cGVHD) but there is currently no meta-analysis to confirm the association. Therefore, we conducted a meta-analysis to evaluate the association between TBI and GVHD.

Method: We performed a comprehensive literature search in MEDLINE, Cochrane Library and EMBASE database from inception to February 2019 for studies that evaluated the association between TBI and aGVHD or cGVHD. We calculated pooled odd ratios (OR) with 95% confidence interval (CI) and I2 statistic using the random-effects model (Odd Ratio) of aGVHD and cGVHD in the TBI group versus the non-TBI group.

Result: Thirteen studies were included involving a total of 19,303 participants (TBI=10,222, non-TBI 9,081).There was a significant association between TBI use and development of aGVHD (pooled OR 1.32, 95% CI 1.09-1.60, I2=76.7 %) and cGVHD (pooled OR 1.32, 95% CI 1.12-1.54, I2=46.2%)

Conclusion: The results of our meta-analysis suggested that the use of TBI is significantly associated with aGVHD and cGVHD in the setting of allogeneic hematopoietic stem cell transplant.

Citation Format: Kittika Poonsombudlert, Chattip Prueksapraopong, Jakrin Kewcharoen, Nath Limpruttidham. Total body irradiation and graft-versus-host disease: A meta-analysis [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 6006.
Abstract 6010: Tumor mhicroenvironment effects on hematopoietic stem cell induced tumor growth following radiation therapy
Authors: Parsons Tyler M. , Buelow Katie , Hanna Alaa , Galoforo Sandra , Wilson George , Madlambayan Gerard
Published: September 2020

The tumor microenvironment (TME), comprised of tumor tissue, stromal cells, extracellular matrix (ECM), bone marrow (BM)-derived immune cells, and various signaling molecules, rarely remains static. These components continuously respond to internal and external stimuli to promote tumor progression as well as regrowth after therapy. Recently, we reported that functional BM-derived hematopoietic stem and progenitor cells (HSPCs) take up residence within growing lung tumors. Interestingly, when tumors were

exposed to radiation therapy (RT) there was a significant increase in HSPC numbers within tumors (p<0.003). These HSPCs seemed to positively affect tumor regrowth as their numbers directly correlated with regrowth rates (R2=.4875). To determine the role of HSPCs in tumor growth we proposed a mechanism wherein tumor-associated HSPCs are maintained within tumors but following exposure to RT can differentiate into tumor-supportive macrophage populations, thus aiding in tumor regrowth. To test this hypothesis, Lewis Lung Carcinoma tumors were subcutaneously generated in mice and exposed to either RT or no radiation treatment (NRT). Analysis using RT-PCR and immunohistochemistry demonstrated that RT produces TME conditions that favor the differentiation of HSPCs into tumor-supportive M2-macrophages through the increased production of colony stimulating factor 1 (CSF-1) (p<0.005). In addition, we show that inhibition of CSF-1R phosphorylation using a pharmacologic inhibitor (GW2580) had no effect on HSPCs numbers but resulted in significantly decreased M2 macrophage numbers resulting in decreased tumor growth and regrowth (p<0.005). Furthermore, we found that the interaction of HSPC-bound integrin α6 with Laminin 5-1-1 is associated with HSPC maintenance within tumors. Interestingly, Laminin 5-1-1 levels were decreased in RT treated tumors compared to NRT controls (p<0.005) further indicating that the RT-derived microenvironments supported HSPC differentiation into M2 macrophages. Overall, these studies define a new understanding of tumor biology in which tumor-associated HSPCs are an essential component of tumor progression and survival and identify new treatment strategies to overcome these effects and achieve more optimal clinical outcomes.

Citation Format: Tyler M. Parsons, Katie Buelow, Alaa Hanna, Sandra Galoforo, George Wilson, Gerard Madlambayan. Tumor mhicroenvironment effects on hematopoietic stem cell induced tumor growth following radiation 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 6010.
Abstract 6012: Cancer stem cells could be responsible for the chimeras of hematopoietic cells in the cancer microenvironment
Authors: Hassan Ghmkin , Afify Said M , Kumon Kazuki , Osman Amira , Zahra Maram H , Seno Akimasa , Seno Masaharu
Published: September 2020

Development of cancer stem cells (CSCs) model from iPSCs for different types of cancers will help to investigate the tumorigenesis and designing new therapies applicable to CSCs. In this context, our lab has developed a novel method to generate CSCs from iPSCs under cancer microenvironment using conditioned medium from different cancer cell lines. In our previous study, BT549 cell line conditioned media was used to convert iPSCs to CSCs (miPSCs-BTcscs). Here, we investigated the potential of adherent CSCs converted from iPSCs to give hematopoietic like cells. Cancer stem cells, miPSCs-BTcscs, were obtained by conversion of miPSCs (iPS MEF-Ng-20D-17) cells in the presence of conditioned medium from breast cancer cell line BT549 cells followed by the primary culture of the tumor formed in Balb/c nude mice. The stemness and tumorigenicity of miPSCs-BTcscs cells had previously been confirmed. miPSCs-BTcscs cells were cultured on gelatin coated dish in absence of Leukemia Inhibitory Factor (LIF) and select undifferentiated cells was done using puromycin. Non-adherent cells (NACs) arising from adherent CSCs collected and their viability, the morphology and the expression of hematopoietic cells markers were investigated using Giemsa staining, flow cytometry, immunofluorescence staining, and RT-PCR. Moreover, to assess the potential of clonogenicity in vivo and the engraftment of NACs, NACs were injected into the tail vein of busulfan conditioned Balb/c nude mice. The round nucleated NACs were found to be viable, positive for Lin markers and CD34 and expressed hematopoietic markers just like homing to the bone marrow. When NACs were injected into a mouse, Wright-Giemsa staining showed that the number of white blood cells got higher than those in the control mice after 4 weeks. As conclusion, CSCs were demonstrated to have the potential to provide progenies with hematopoietic markers, morphology and homing ability to bone marrow, which could give a new insight into tumor microenvironment according to the plasticity of CSCs.

Citation Format: Ghmkin Hassan, Said M Afify, Kazuki Kumon, Amira Osman, Maram H Zahra, Akimasa Seno, Masaharu Seno. Cancer stem cells could be responsible for the chimeras of hematopoietic cells in the cancer microenvironment [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 6012.
Abstract 6016: The impact of cancer stem cell markers in the tissues of pancreatic ductal adenocarcinoma for prognosis after surgical resections
Authors: Inokawa Yoshikuni , Yamada Suguru , Hayashi Masamichi , Sunagawa Yuki , Sonohara Fuminori , Takami Hideki , Hattori Norifumi , Kanda Mitsuro , Kobayashi Daisuke , Tanaka Chie , Nakayama Goro , Koike Masahiko , Kodera Yasuhiro
Published: September 2020

Introductions: The prognosis after resection of pancreatic ductal adenocarcinoma (PDAC) is still limited though the survival benefit is recently reported by neo-adjuvant chemotherapy (NAC). The existence of cancer stem cells (CSCs) is known as one of the important mechanisms for cancer cells to survive against chemotherapy. We hypothesized that the population of CSCs among cancer cells after chemotherapy could contribute recurrence after surgical resections, and could be detected by some CSC markers.

Methods: We checked elevation of CSC markers in resected cases after pre-operative therapy (NAC with gemcitabine plus nab-paclitaxel, n=3; NAC with FOLFIRINOX, n=3; and neo-adjuvant chemo-radiotherapy (NACRT) with S1+Radiation, n=3) compared to surgery alone (n=3) with RT2 Profiler PCR Array. As CSC models, we performed sphere formation assay with PDAC cell lines (Panc1 and BXPC3) and evaluated the expression of CSC markers with qRT-PCR. Then, we performed qRT-PCR with 48 PDAC resected cases including 26 cases with pre-operative therapy (54%).

Results: RT2 Profiler PCR Array showed higher expression of some CSC markers with pre-operative treatment (n=9) than without them (n=3). Spheroid assay showed elevation of CSC markers in spheroid compared to parental cells (Panc1: 10 among 16 markers, BXPC3 14 among 16 markers). In the cohort of 48 resected PDAC patients, ALDH1, Nanog, SOX2, MYC CXCR4 and DCLK1 showed significantly higher expression in pre-operative therapy than in surgery alone (P<0.05). About prognosis, higher expression of YAP1 in tumor tissues showed worse outcomes of recurrence free survival (RFS) (P=0.06), and higher CXCR4 in adjacent parenchyma of pancreas showed worse RFS among patients with NAC and NACRT (P=0.08).

Conclusions: Our findings indicated that CSCs are more in resected tissues after NAC and NACRT than in without pre-operative therapy. YAP1 expression in tumors or CXCR4 in adjacent parenchyma of pancreas can be prognosis markers after surgical resection for PDAC.

Citation Format: Yoshikuni Inokawa, Suguru Yamada, Masamichi Hayashi, Yuki Sunagawa, Fuminori Sonohara, Hideki Takami, Norifumi Hattori, Mitsuro Kanda, Daisuke Kobayashi, Chie Tanaka, Goro Nakayama, Masahiko Koike, Yasuhiro Kodera. The impact of cancer stem cell markers in the tissues of pancreatic ductal adenocarcinoma for prognosis after surgical resections [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 6016.
Abstract 6018: The perfused 3DKUBE™ rare tumor assay models in vivo drug response
Authors: Millard Melissa , Appleton Kathryn M. , Elrod Ashley , Bateman Nicholas W. , Abulez Tamara , Conrads Kelly , Hood Brian , Conrads Thomas P. , Holmes Lillia M. , DesRochers Teresa M.
Published: September 2020

Models that accurately reflect patient drug response are essential for the clinical design of personalized treatment plans and necessary for preclinical drug development. The advancement of predictive models for rare tumor types is impeded in part, by the relative scarcity of fresh tumor tissue available for study. To address the problem of tissue availability we have developed a label-free, combined functional and chemical selection method for the isolation of rare tumor cancer stem cells (CSC) and circulating tumor cells (CTC) from primary patient tissue and blood. Enriched cells were expanded as 3D microtumors under optimized conditions, validated as CSC through in vivo tumorigenesis studies, and characterized by correlative genomic, proteomic/phosphoproteomic, and phenomic analysis. We found isolation and expansion by this method yielded a source of primary cells suitable for live, cell-based predictive drug screening in multiple rare tumor derived models of neuroendocrine and mesenchymal origin, including locally or regionally advanced and metastatic SCLC, recurrent Merkel Cell Carcinoma, recurrent osteosarcoma and dermatofibrosarcoma protuberans. The 3DKUBE™ rare tumor assay was performed using validated CSCs cultured as perfused 3D microtumors (pMTs). Drug studies using the perfused, 3DKUBE™ rare tumor assay modeled individual patient response better than CSC-based or 3D static microtumor-based drug screens and thus demonstrate the effectiveness of this platform for predictive modeling of individual patient drug response. Taken together, this system provides a means of performing ex vivo drug response experiments on very small tissue samples, including core biopsies, with relevant results for patients.

Citation Format: Melissa Millard, Kathryn M. Appleton, Ashley Elrod, Nicholas W. Bateman, Tamara Abulez, Kelly Conrads, Brian Hood, Thomas P. Conrads, Lillia M. Holmes, Teresa M. DesRochers. The perfused 3DKUBE™ rare tumor assay models in vivo drug response [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 6018.
Abstract 6024: An integrin αvβ3+ stem-like tumor cell subset promotes breast cancer progression via c-Jun/AXL signaling
Authors: Sun Qi , Wang Yufen , Officer Adam , Harismendy Olivier , Desgrosellier Jay S.
Published: September 2020

Breast cancer cells bearing similarities to normal mammary stem cells play a key role in tumor progression. Here, we characterize a unique stem-like tumor cell population that expresses the integrin αvβ3 and is associated with aggressive patient disease. We now show that αvβ3+ cells that also express EpCAMLow represent a functionally and transcriptomically distinct breast cancer cell type. Rare αvβ3+ cells enrich for stemness properties such as anchorage-independent colony formation, self-renewal, differentiation and tumor initiation at limiting dilution. Unbiased whole transcriptome analysis by RNA-Seq identified several genes specifically upregulated in αvβ3+ cells that are also enriched in normal adult mammary stem cells. Phospho-array profiling of major signaling pathways revealed increased activation of the proto-oncogene c-Jun in αvβ3+ stem-like cells. Inhibition of JNK/c-Jun signaling further identified the receptor tyrosine kinase AXL as a critical downstream target gene necessary for tumorsphere formation. Our findings reveal a c-Jun/AXL signaling pathway required for stemness properties in αvβ3+ tumor cells important for breast cancer progression.

Citation Format: Qi Sun, Yufen Wang, Adam Officer, Olivier Harismendy, Jay S. Desgrosellier. An integrin αvβ3+ stem-like tumor cell subset promotes breast cancer progression via c-Jun/AXL signaling [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 6024.
Abstract 604: Combination of the Aurora A inhibitor TAS-119 and adavosertib results in synergistic c-Jun-mediated cell death in head and neck and lung cancers
Authors: Lee Jong Woo , Yang Cindy , Burtness Barbara
Published: September 2020

Head and neck squamous cell carcinomas (HNSCC) account for more than 800,000 cases annually worldwide. Human papillomavirus (HPV)-negative HNSCC mainly harbor disruptive mutations in TP53 and/or CDKN2A tumor suppressor genes, which are associated with refractoriness to treatment as well as poor prognosis and survival. Synthetic lethal approaches hold promise for such patients. Cancers with TP53/CDKN2A mutations lose the G1/S checkpoint, and foster a dependence on the G2/M checkpoint as the primary mechanism to repair the resulting accumulation of genomic instability, thus creating a vulnerability that can potentially be exploited with synthetic lethal targeted therapy. We have previously demonstrated in HPV-negative HNSCC patients that elevation in expression of Aurora kinase A (AURKA), a crucial mitotic regulator, is correlated with worse outcome. Combined inhibition of AURKA and WEE1, a G2/M checkpoint kinase, resulted in a synergistic anti-tumor effect in HNSCC in vitro and in vivo. Upon these findings, we confirmed synergy in testing the more specific AURKA inhibitor, TAS-119 in HNSCC and lung cancers harboring TP53 mutations, and explored mechanisms of cell death. TAS119 and adavosertib combination treatment synergistically suppressed cell viability, clonogenic cell survival, anchorage-independent cell growth in soft agar and oncosphere formation relative to vehicle and single-agent treatment in HNSCC FaDu and CAL27 cells, and lung cancer NCI-H520 and NCI-H358 cells bearing TP53/CDKN2A mutations. Of note, this synergistic anti-tumor effect was not seen in normal human tracheobronchial epithelial cells, predicting a favorable therapeutic index. Furthermore, combination treatment accelerated an accumulation of mitotic catastrophe with mitotic arrest, and led to apoptotic cell death. Mechanistically, combination TAS-119 and adavosertib in HNSCC cells drastically augmented DNA damage and phosphorylation of c-Jun (Ser 63) as demonstrated by kinase array and western blotting, indicating an activation of c-Jun-mediated pro-apoptotic genes. In vivo, this combination demonstrated a synergistic suppressive effect on tumor growth in FaDu xenografts compared with either vehicle or single-agent treatment. Taken together, these results support the promise of further clinical evaluation of AURKA inhibitor in combination with WEE1 inhibitor as a novel and effective treatment for HNSCC and lung cancer patients with TP53/CDKN2A mutations.

Citation Format: Jong Woo Lee, Cindy Yang, Barbara Burtness. Combination of the Aurora A inhibitor TAS-119 and adavosertib results in synergistic c-Jun-mediated cell death in head and neck and lung cancers [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 604.
Abstract 6055: Discovery of epitope-selective anti-CD25 targeting therapeutic antibodies for effective Treg cell depletion in cancer using a novel AI/ML based platform
Authors: Gip Phung , Li Bing , Estelles Angeles , Daino-Laizure Hanako , Gurinovich Oleg , Zalicki Piotr , Hauser Kevin , Taguchi Alex , Barrett Christian , Morin Andrew , Willis Jordan , Srinivasan Mohan , Bright Isaac , Greving Matt
Published: September 2020

Therapeutic antibodies have contributed significantly to better clinical outcomes across multiple tumor types. However, there is still unmet clinical need in cancers with an immunosuppressive and stroma rich tumor microenvironment (TME). One key limitation in early antibody discovery is immunodominance - an evolutionary bias toward specific epitopes that steer antibody selection away from potentially high-efficacy epitopes. Our integrated computational and laboratory approach to antibody discovery enabled us to build focused epitope embodiments of structural epitopes. These epitope embodiments, called Meso-scale Engineered Molecules (MEMs), are used in antibody selection to steer hits toward intended epitopes. CD25 (IL2Rα) is highly expressed on regulatory T-cells (Tregs) that drive immunosuppression in the TME. Depletion of Tregs can restore anti-tumor T effector (Teff) function in the TME, by increasing the Teff/Treg ratio, especially when combined with checkpoint-inhibitors. Using our epitope-selective antibody discovery platform, we built MEMs that embody eight CD25 structural epitopes outside the CD25:IL-2 interface. Through in vitro selection, several high-affinity (median KD = 25 nM) anti-CD25 antibodies from each of the eight intended CD25 epitopes were identified and confirmed to be on epitope by cross-blocking assays and in-epitope alanine mutations. Several conventional- and epitope-selective anti-CD25 clones were converted to human IgG1 and tested in in vitro assays. As a result, these antibodies bound specifically to CD25+ cells, preserve IL-2 signaling via pSTAT5 assays and elicited potent ADCC activity using human effector cells, superior to benchmark antibodies. These studies show that our integrated computational and laboratory platform improves the antibody discovery process, producing clones with biological function and improved therapeutic efficacy.

Citation Format: Phung Gip, Bing Li, Angeles Estelles, Hanako Daino-Laizure, Oleg Gurinovich, Piotr Zalicki, Kevin Hauser, Alex Taguchi, Christian Barrett, Andrew Morin, Jordan Willis, Mohan Srinivasan, Isaac Bright, Matt Greving. Discovery of epitope-selective anti-CD25 targeting therapeutic antibodies for effective Treg cell depletion in cancer using a novel AI/ML based platform [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 6055.