bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3649: Allelic imbalance in KMT2A-rearranged infant acute lymphoblastic leukemia
Authors: Yoo Byunggil , Farooqi Midhat S. , Kostadinov Rumen , Cheung Warren , Farrow Emily , Kelley Shannon , Miller Neil , Ge Bing , Gibson Margaret , Brown Patrick , Guest Erin M. , Pastinen Tomi
Published: August 2019

Background

Infant acute lymphoblastic leukemia (ALL) is a malignant disorder with poor clinical outcome. It is well-known that infant ALL cases with rearrangement of the KMT2A gene (KMT2A-r) have an even poorer prognosis than non-KMT2A-r cases. Interestingly, KMT2A-r infant ALL cases have remarkably few other genetic alterations. We hypothesized that non-coding events in cancer genomes (e.g. loss of expression or methylation) may play a role in this disease. Such events can be captured using genomic analyses in haploid genomes using analytical approaches for allelic imbalance quantification. Here, we examine whether allelic imbalance is a feature of infant ALL.

Methods

We performed whole genome sequencing (WGS) and RNA sequencing on peripheral blood or bone marrow specimens from 29 KMT2A-r cases and 14 non-KMT2A-r cases at diagnosis (DX), remission (MD), and relapse (RL) as applicable. WGS data from MD samples was phased using a 1000 Genomes reference panel. Biallelic expression was measured on the phased genome for transcripts with at least 2 SNPs and at least 15 aligned reads. Lesser allele fraction (LAF; allele fraction for the less prevalent allele) for DX/RL versus MD samples was compared for transcripts that had LAFs available in at least 3 cases using t-test. Transcript-level p-values were aggregated at the gene level using the Sidak method.

Results

Allelic imbalance in expression (LAF <= 0.2) was observed in an average of about 600 genes per sample in infant ALL regardless of timepoint. Disease-specific allelic imbalance (skewed in DX samples but not in paired MD samples) was detected in 431 genes for the KMT2A-r cohort and 77 genes for the non-KMT2A-r cohort. A total of 38 genes with allelic imbalance were shared between the two cohorts. Notably, KMT2A was observed to be imbalanced in KMT2A-r samples. Genes of known significance that were found to be skewed included HOXA9 and PARP8.

Discussion

Our study suggests that allelic imbalance quantification may help uncover novel molecular mechanisms in infant ALL, especially KMT2A-r cases. However, similar to gene expression, the patterns of allelic imbalance in KMT2A-r cases at DX do not allow efficient prediction of which patients go on to relapse.

Citation Format: Byunggil Yoo, Midhat S. Farooqi, Rumen Kostadinov, Warren Cheung, Emily Farrow, Shannon Kelley, Neil Miller, Bing Ge, Margaret Gibson, Patrick Brown, Erin M. Guest, Tomi Pastinen. Allelic imbalance in KMT2A-rearranged infant acute lymphoblastic leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3649.
Abstract 3676: Novel tumor suppressive role of Peptidylarginine deiminase IV involving cancer stem cell regulation in breast cancer models
Authors: Ochoa Humberto J. , Moshkovich Nellie , Tang Binwu , Yang Howard H. , Lee Maxwell P. , Wakefield Lalage M.
Published: August 2019

Breast cancer accounts for >500,000 deaths annually worldwide. Despite therapeutic advances, treated patients often relapse with metastases. Emerging research indicates that relapse may be driven by a subpopulation of tumor cells termed cancer stem cells (CSCs), that are uniquely capable of self-renewal and have enhanced drug resistance. Understanding the properties of these CSCs is critical for development of more effective therapies. Peptidyl arginine deiminase 4 (PADI4) is an enzyme that catalyzes deimination of arginine to citrulline. Citrullination of histones modulates expression of key stem cell transcription factors in embryonic stem cells (Christophorou, et al., 2014, Nature 507:104), so we hypothesized that PADI4 could be involved in CSC regulation. We showed that PADI4 mRNA and protein are expressed at varying levels across a panel of breast cancer cell lines, and we knocked down PADI4 in two high expressing lines, MCF10Ca1h and MDAMB231-LM2. PADI4 knockdown did not alter tumor cell proliferation in vitro, but it enhanced migration and invasion and increased cell survival. Importantly, PADI4 knockdown increased clonogenicity and enhanced tumorsphere formation, suggesting that it specifically increases the CSC population. In vivo studies showed higher tumor initiation efficiency following PADI4 knockdown, and increased lung metastasis. PADI4 is predicted to have multiple isoforms with unknown biological activity that complicate PADI4 expression-based analysis in patient data sets. We circumvented this issue by pharmacologically inhibiting PADI4 in MCF10Ca1h to generate a transcriptomic signature reflecting PADI4 gene activity. This PADI4 gene activity signature was shown to correlate with lower tumor grade and better disease outcome in transcriptomic datasets from human ER+ breast cancers. In conclusion, our findings suggest that PADI4 functions as a tumor suppressor in breast cancer, in part through effects on the CSC.

Citation Format: Humberto J. Ochoa, Nellie Moshkovich, Binwu Tang, Howard H. Yang, Maxwell P. Lee, Lalage M. Wakefield. Novel tumor suppressive role of Peptidylarginine deiminase IV involving cancer stem cell regulation in breast cancer models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3676.
Abstract 3684: The TGFβ2-Snail1-miRNATGFβ2 circuitry is critical for the development of aggressive functions in breast cancer
Authors: He Zhimin , Zheng Guopei , Zhang Zhijie , Liu Hao , Zhou Binhua Peter
Published: August 2019

There have been contradictory reports on the biological role of transforming growth factor-βs (TGFβs) in breast cancer, especially with regard to their ability to promote epithelial-mesenchymal transition. Here, we show that TGFβ2 is preferentially expressed in mesenchymal-like breast cancers and maintains the epithelial-mesenchymal transition phenotype, correlating with cancer stem cell -like characteristic, growth, metastasis and chemo-resistance and predicting worse clinical outcome. However, this is only true in ERα- breast cancer. In ERα+ luminal-type breast cancer, estrogen receptor interacts with p-Smads to block TGFβ signaling. Furthermore, we also identify a microRNAs (miRNAs) signature (miRNAsTGFβ2) that is weakened in TGFβ2-overexpressing breast cancer cells. We discover that TGFβ2-Snail1 recruits EZH2 and BMI1 to convert miRNAsTGFβ2 promoters from an active to repressive chromatin configuration and then repress miRNAsTGFβ2 transcription, forming a negative feedback loop. On the other hand, miRNAsTGFβ2 overexpression reverses the mesenchymal-like traits in agreement with inhibition of TGFβ2-Snail1 signaling in breast cancer cells. These findings clarify the roles of TGFβ2 in breast cancer and suggest novel therapeutic strategies based on the TGFβ2-Snail1-miRNATGFβ2loop for a subset type of human breast cancers.

Keywords: TGFβ, EMT, miRNA, Snail1, feedback loop This study was supported by grants from the National Natural Science Foundation of China (No.81672616, 81872197, 81402196, 81272450 and 81772825); Guangdong Natural Science Funds for Distinguished Young Scholars (2016A030306003); Guangdong Special Support Program (2017TQ04R809).

Note: This abstract was not presented at the meeting.

Citation Format: Zhimin He, Guopei Zheng, Zhijie Zhang, Hao Liu, Binhua Peter Zhou. The TGFβ2-Snail1-miRNATGFβ2 circuitry is critical for the development of aggressive functions in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3684.
Abstract 3698: Conditionally reprogrammed cells from patient-derived xenograft to model neuroendocrine prostate cancer development
Authors: Ci Xinpei , Hao Jun , Dong Xin , Xue Hui , Wu Rebecca , Haegert Anne M. , Collins Colin C. , Lin Dong , Wang Yuzhuo
Published: August 2019

Treatment-emergent neuroendocrine prostate cancer (t-NEPC) is a lethal subtype of advanced prostate cancer that occurs via NE transdifferentiation of prostate adenocarcinomas in response to androgen receptor (AR)-inhibition therapy. Study of t-NEPC has been hampered by a lack of clinically relevant models. We previously established a unique and first-in-field patient-derived xenograft (PDX) model of adenocarcinoma (LTL331)-to-NEPC (LTL331R) transdifferentiation. In this study, we established conditionally reprogrammed (CR) cells from the adenocarcinoma PDX tumor line LTL331. These LTL331-derived CR (LTL331-CR) cells retained the same genomic mutations of the parental tumor and, can be genetically manipulated and continuously propagated in vitro. Further androgen deprivation treatment on LTL331-CR cells showed no effect on cell proliferation. Transcriptomic analyses of the LTL331-CR cells revealed profound downregulation of androgen response pathway, and enrichment of stem/progenitor-like marker genes, compared with the parental tumor LTL331. Notably, when grafted back into the subrenal capsule of male NOD/SCID mice, these LTL331-CR cells gave rise to NEPC tumors directly as manifested by histological expression of NE markers. Transcriptomic analyses of the newly developed NEPC tumors also demonstrated marked enrichment of NEPC signature genes and loss of AR signaling genes. This study provides a novel strategy to investigate the mechanisms underlying t-NEPC development with a unique PDX by enabling gene manipulation ex vivo and subsequent functional evaluation in vivo.

Citation Format: Xinpei Ci, Jun Hao, Xin Dong, Hui Xue, Rebecca Wu, Anne M. Haegert, Colin C. Collins, Dong Lin, Yuzhuo Wang. Conditionally reprogrammed cells from patient-derived xenograft to model neuroendocrine prostate cancer development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3698.
Abstract 3700: Investigating the therapeutic potential of parthenolide for hematopoietic neoplasms in dogs
Authors: Schlein Lisa J. , Rose Barbara , Peterson Aubree , Thamm Douglas
Published: August 2019

The purpose of this research is to explore the therapeutic potential of parthenolide (PTL) to treat various hematopoietic neoplasms in dogs; additionally, some dog breeds are enriched for development of mast cell neoplasia and histiocytic sarcoma, providing translational study populations for rare and deadly human diseases. Growth inhibition assays were performed using a panel of canine mast cell, histiocytic sarcoma, lymphoma, and leukemia cell lines, with PTL alone or in combination with redox-perturbing standard-of-care therapeutics. Cell death was assessed using flow cytometry. Immunofluorescence and immunoblotting were used to assess NFκB localization and phosphorylation, respectively. All immortalized canine cell lines evaluated are sensitive to PTL therapy and undergo dose-dependent apoptosis following exposure to drug. PTL exposure leads to inhibition of NFκB, as evidenced by immunofluorescent nuclear exclusion and decreased p65 phosphorylation. Preliminary studies indicate that some standard-of-care therapeutics synergize with PTL. These initial studies show that PTL is a promising therapeutic for hematopoietic neoplasms in dogs. Murine modeling and investigation of redox perturbance are underway and will further investigate PTL’s potential in the clinical setting.

Citation Format: Lisa J. Schlein, Barbara Rose, Aubree Peterson, Douglas Thamm. Investigating the therapeutic potential of parthenolide for hematopoietic neoplasms in dogs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3700.
Abstract 3712: Proliferative and invasive colorectal tumors in pet dogs provide unique insights into human colorectal cancer
Authors: Wang Jin , Wang Tianfang , Sun Yanfang , Feng Yuan , Kisseberth William C. , Henry Carolyn J. , Mok Irene , Lana Susan E. , Dobbin Kevin , Northrup Nicole , Howerth Elizabeth W. , Zhao Shaying , Cai Houjian
Published: August 2019

Spontaneous tumors in pet dogs represent a valuable but undercharacterized cancer model. To better use this resource, we performed an initial global comparison between proliferative and invasive colorectal tumors from 20 canine cases and evaluated their molecular homology to human colorectal cancer (CRC). We sequenced 15 canine intestinal samples for WGS and 26 for RNA-seq. We investigated alterations in the genome and transcriptome, using state-of-the-art analysis tools. We aslso investigated their microbiome, by mapping WGS and RNA-seq read pairs that could not be placed onto the canine genome to two microbial genome databases: The Human Microbiome Project database and all bacterial genomic sequences. We performed the same analysis with TCGA (The Cancer Genome Atlas) colon cancer data. Based on our analysis, proliferative canine tumors harbor overactivated WNT/β-catenin pathways and recurrent CTNNB1 (β-catenin) mutations S45F/P, D32Y and G34E. Invasive canine tumors harbor prominent fibroblast proliferation and overactivated stroma. Both groups have recurrent TP53 mutations. We observed three invasion patterns in canine tumors: collective, crypt-like and epithelial-mesenchymal transition (EMT). We detected enriched Helicobacter bilis and Alistipes finegoldii in proliferative and crypt-like tumors, but depleted mucosa-microbes in the EMT tumor. Additionally, guided by our canine findings, we classified 79% of 478 human colon cancers from TCGA into four subtypes: primarily proliferative, or with collective, crypt-like or EMT invasion features. Their molecular characteristics match those of canine tumors. We showed that consensus molecular subtype 4 (mesenchymal) of human CRC should be further divided into EMT and crypt-like subtypes, which differ in TGF-β activation and mucosa-microbe content. Our canine tumors share the same pathogenic pathway as human CRCs. Dog-human integration identifies three CRC invasion patterns and improves CRC subtyping.

Citation Format: Jin Wang, Tianfang Wang, Yanfang Sun, Yuan Feng, William C. Kisseberth, Carolyn J. Henry, Irene Mok, Susan E. Lana, Kevin Dobbin, Nicole Northrup, Elizabeth W. Howerth, Shaying Zhao, Houjian Cai. Proliferative and invasive colorectal tumors in pet dogs provide unique insights into human colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3712.
Abstract 3715: Cancer as a consequence of breaking through evolutionary constraints on longevity
Authors: Modiano Jaime F. , Sarver Aaron L.
Published: August 2019

It is well known that the risk of cancer increases with age, but when viewed through a comparative lens, this risk is species specific and exists at a relatively low background in the animal kingdom. The exceptions, where cancer is a major cause of mortality, are humans and companion dogs and cats. Indeed, it is estimated that more than 50% of pet dogs that reach the age of 10-years will die from cancer. Colloquially, there is a perception that there is a “cancer epidemic” in dogs caused by exposures to real or perceived mutagens, but this hypothesis is based on presumptions for which supporting evidence is weak or absent. Here, we propose an alternative hypothesis, that replacing natural selection with artificial selection for form and function, and superimposing longevity that breaks the evolutionary constraints for that species inevitably creates cancer-prone phenotypes. Modern pet dogs have achieved a social status equivalent to human family members. With this have come better health care, reduced early mortality, and greater longevity. Although dogs were first domesticated15,000 to 20,000 years ago and breeds were derived over the last 200 to 400 years, significant gains in longevity and the recognition of cancer as a major cause of mortality in pet dogs have taken place over the past 40 to 60 years, coinciding with the transition of dogs from working animals to pets. Breaking through the evolutionary constraint on longevity comes at a price: long-lived species such as elephants, whales, mole rats, and others have acquired a diversity of cancer-protective mechanisms, achieving a balance between longevity and their reproductive and energetic investments over tens of millions of years. But no such cancer-protective mechanisms have been identified in dogs, or for that matter in humans. In both species, risk increases with age, but in dogs, risk is magnified by artificial selection. Osteosarcoma provides an example that can be generalized. The underlying causal factor is selective breeding for large size, but this is only able to manifest itself because modern pet dogs exceed their evolutionarily expected lifespan by at least 2 or 3 times. We conclude that appreciating the role of longevity as a major cause of cancer in pet dogs, as well as in other model organisms will improve our ability to design and interpret experiments that will show greater fidelity when translated to humans, where the same risk factors are operative, although they are partly masked by the influence of behavioral or occupational exposures to strong mutagens.

Citation Format: Jaime F. Modiano, Aaron L. Sarver. Cancer as a consequence of breaking through evolutionary constraints on longevity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3715.
Abstract 3901: Nonclinical safety evaluation of CA102N, a hyaluronic acid (HA) and nimesulide (Nim) conjugate, for colorectal cancer treatment
Authors: Tchaparian Eskouhie , Lin Louis , Hsu Elin , Chen Ivy , Wu Albert
Published: August 2019

CA102N is a conjugate of the biological polymer hyaluronic acid (HA) and Nimesulide (Nim), a cyclooxygenase-2 (COX-2) inhibitor that represents a promising new approach to addressing cancer treatment. Similar to an antibody-drug conjugate or a pro-drug, CA102N enhances the efficacy of Nim through targeting and binding of HA to CD44 receptors highly expressed in solid tumors. Pharmacological (in vitro and in vivo) studies indicated that CA102N exhibits apoptotic and tumor growth inhibitory activities against colorectal cancer. The MOA of CA102N was demonstrated to be related to apoptosis and disruption of the PI3K/Akt/mTOR pathway in addition to inhibition of the COX-2/PGE2 related pathways.

Safety of CA102N was assessed in acute dose range finding and repeat-dose intravenous toxicity studies in rats and beagle dogs. Three dose levels including the Maximum Tolerated Dose (MTD) from DRF study, a middle dose and a low dose equivalent to the proposed clinical dose were administered twice weekly (BID). A 28-day recovery group was also included in the rat study.

Overall CA102N was well tolerated in both species. CA102N did not induce any significant effects on the CNS, the cardiovascular and respiratory systems in animal models. Some changes in hematological parameters were observed only in rats at the highest dose (Std 10: ≥ 600 mg/kg/day ), however, most of these observations were reversible. No hERG inhibition (in vitro) was observed suggesting the risk of QT prolongation with CA102N is low. The Highest Non-Severely Toxic Dose (HNSTD) of CA102N in the dog was determined to be 280 mg/kg/day (7.24 mg/kg in Nim equivalents ) and the rat NOAEL was 400 mg/kg/day (10.16 mg/kg in Nim equivalents ). These doses are 2.3 times (rat) and 5.6 times (dog) of nimesulide dose to be evaluated (up to 0.72 mg/kg of nimesulide) in the proposed phase 1 study.

On the basis of the mechanism of action and the nonclinical safety profiles, toxicity associated risks to patients appear to be low at the proposed clinical dose levels. A Phase I study of CA102N in patients with locally advanced or metastatic colorectal cancer is ongoing.

Citation Format: Eskouhie Tchaparian, Louis Lin, Elin Hsu, Ivy Chen, Albert Wu. Nonclinical safety evaluation of CA102N, a hyaluronic acid (HA) and nimesulide (Nim) conjugate, for colorectal cancer treatment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3901.
Abstract 3908: The combination of NVP-BEZ235 & endostatin exerts synergistic anticancer activity against triple-negative breast cancer in vitro and in vivo
Authors: Wang Xiaoshan , Zhang Xiao , Li Fanghua
Published: August 2019

Background: Triple-negative breast cancer(TNBC) take up 15% of invasive breast cancer. TNBC has no effective molecular targets & unfavourable prognosis. We investigated the effects of NVP-BEZ235, a novel dual PI3K/mTOR inhibitor, a combine with or not an angiogenesis inhibitor Endostatin in the orthotopic TNBC model & explored its anti-tumor & anti-angiogenesis mechanisms.

Material & Methods: Assess sensitivity of the cells to the drug by MTT assays.Orthotopic xenografts were performed for tumor growth and survival studies. Additionally, Immunohistochemistry for VEGF and MVD(CD34) was performed using the EnVision/HRP technique,Serum VEGF was detected by ELISA method.

Results: BEZ235 effectively inhibited cell proliferation in vitro & provided additive effects when combine with Endostatin.Treatment with BEZ235 & Endostatin resulted in inhibition of tumor growth & prolongation of survival time of tumor-bearing mice in vivo.Immunohistochemical analysis revealed that intratumoral proliferation & angiogenesis was significantly suppressed. Furthermore, Changing of the number of Serum VEGF support the finding of angiogenesis inhibition too.

Conclusion: Our findings suggest that BEZ235 exerts antitumor effects against TNBC & enhances effects of Endostatin through inhibition of cell proliferation & tumor angiogenesis. This approach may represent a promising combination targeted therapy for TNBC treatment.

Note: This abstract was not presented at the meeting.

Citation Format: Xiaoshan Wang, Xiao Zhang, Fanghua Li. The combination of NVP-BEZ235 & endostatin exerts synergistic anticancer activity against triple-negative breast cancer in vitro and in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3908.
Abstract 393: Deoxymab: A targeted biologic that is synthetically lethal to TNBC brain metastases
Authors: Zhang Shenqi , May Christopher , Shirali Anupama , Dubljevic Valentina , Campbell James A. , Zhou Jiangbing , Hansen James E.
Published: August 2019

Brain metastases in triple-negative breast cancer (TNBC) harbor defects in homology-directed DNA repair that may be exploited to develop new therapeutic strategies. However, these tumors are protected by the blood-brain barrier (BBB), and mechanisms of DNA repair are sequestered in the nucleus. Methods to cross the BBB and target intranuclear processes are needed.

3E10 is a nuclear-penetrating lupus anti-DNA autoantibody that inhibits the DNA damage response to selectively kill cancer cells with defects in DNA repair. 3E10 localizes to DNA in tumor environments, and penetrates cells via the ENT2 equilibrative nucleoside transporter. ENT2 is expressed in the BBB, and 3E10 has previously delivered cargo proteins to the brain. We believe 3E10 can be turned against TNBC brain metastases, and used MDA-MB-231-BR brain-seeking human TNBC cells and tumors as a model system to test this hypothesis.

We re-engineered 3E10 into an optimized fragment, called Deoxymab-1 (DX1), to maximize effect on cancer cells and minimize toxicity. DX1 expressed in CHO cells was purified over a HiTrap Capto S column by FPLC, and purity and quality was confirmed by SDS-PAGE and SEC-HPLC. Immunostaining and colony formation assays demonstrated that DX1 penetrated the nuclei of ~100% of MDA-MB-231-BR cells in culture and was synthetically lethal to the cells, with doses of 5 and 10 μM reducing surviving fractions to 0.47±0.15 (P<0.03) and 0.29±0.07 (P<0.01), respectively.

TNBC brain metastases were established in nude mice by intracardiac injection of 1.75x105 MDA-MB-231-BR cells transfected with luciferase. One week later the presence of brain metastases was confirmed by IVIS. Mice were treated with tail vein injection of vehicle control (n=7) or DX1 (20 mg/kg) (n=7) 3x/week for 3 weeks. Tumor burden was monitored by weekly IVIS. DX1 significantly suppressed growth of the brain metastases (Table 1). These results establish proof of the novel concept of use of a modified lupus autoantibody against TNBC brain metastases.

Table 1:Brain Radiance Efficiencies (x105)TreatmentWeek 0Week 1Week 2Week 3Control1.2±0.22.5±0.45.1±0.720±5DX11.3±0.21.4±0.32.0±0.23.8±0.8Pns<0.01<0.01<0.02

Citation Format: Shenqi Zhang, Christopher May, Anupama Shirali, Valentina Dubljevic, James A. Campbell, Jiangbing Zhou, James E. Hansen. Deoxymab: A targeted biologic that is synthetically lethal to TNBC brain metastases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 393.
Abstract 3944: MKI-1 is a small molecule inhibitor of MASTL with antitumor and radiosensitizer activity in breast cancer cells
Authors: Kim Ah-Young , Kim Jae-Sung
Published: August 2019

Although microtubule-associated serine threonine kinase like (MASTL) is an emerging target for selective anticancer treatment, a small-molecule inhibitor of MASTL with antitumor activity had not been reported yet. This study showed the new MASTL inhibitor, MKI 1 (MASTL Kinase Inhibitor 1), with antitumor activity and radiosensitizing effect in breast cancer in vitro and in vivo. MKI 1 was identified and validated its inhibitory activity to MASTL by in silico and in vitro analysis, respectively. MKI 1 inhibited phosphorylated ENSA, a direct substrate of MASTL, but not the other AGC kinases such as Akt signaling in breast cancer cells. In addition, MKI 1 reduced various oncogenic properties of breast cancer cells rather than normal breast cells. Furthermore, MKI 1 increased the radiosensitivity of breast cancer cells and breast cancer stem cells in response to irradiation. We also confirm the in vivo antitumor activity and radiosensitizing effects of MKI 1 in breast tumor xenograft model. Therefore, our results provide the new line of a small-molecule inhibitor of MASTL, MKI 1, with the antitumor and radiosensitizer activity in breast cancer in vitro and in vivo.

Citation Format: Ah-Young Kim, Jae-Sung Kim. MKI-1 is a small molecule inhibitor of MASTL with antitumor and radiosensitizer activity in breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3944.
Abstract 3963: Deep serial analysis of the TCR/BCR CDR3 adaptome repertoire in peripheral blood of pancreatic cancer patients in a randomized study of neoadjuvant chemotherapy with or without autophagy inhibition
Authors: Wang Yue , Lotze Michael T.
Published: August 2019

Pancreatic cancer has a paucity of dendritic cells and almost no T lymphocytes. Here, we used Next Generation Sequencing following dimer avoidance multiplexed PCR (DAM-PCR) immune repertoire to study the characteristics and diversity of the TCR repertoire among PDAC (Pancreatic Ductal Adenocarcinoma) patients who underwent chemotherapy with or without Hydroxycholoroquine followed by surgical extirpation. We sequenced the TCR α,β,γ, and δ chains and the B-cell receptor (BCR) heavy chain (IgH) complementary-determining region 3 (CDR3) repertoire in peripheral blood mononuclear cells (PBMCs) from 10 PDAC patients before (Pre-chemo) and after chemotherapy (Post-chemo) and following operation (Postop). The distribution of CDR3, VD indel, and DJ indel lengths were similar among the Prechemo, Postchemo and Postop groups. The TCR repertoire diversity of PDAC patients was much lower compared to healthy donors (HD). The diversity index (D50) of TCR α and β’s in PDAC patients who received HCQ during chemotherapy was less than in patients without HCQ (p<.05). TRBV expression increased and some public sequences diminished at individual time points following chemotherapy and postop. Full analysis of an ongoing study will be presented.

Citation Format: Yue Wang, Michael T. Lotze. Deep serial analysis of the TCR/BCR CDR3 adaptome repertoire in peripheral blood of pancreatic cancer patients in a randomized study of neoadjuvant chemotherapy with or without autophagy inhibition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3963.
Abstract 3966: Combined Rho-kinase inhibition and immunogenic cell death triggers and propagates immunity against cancer
Authors: Kim In-San , Kim Gi Beom , Nam Gi-Hoon , Hong Yeonsun
Published: August 2019

Activation of T-cell immune response is critical for the therapeutic efficacy of cancer immunotherapy. Current immunotherapies have shown remarkable clinical success against several cancers; however, significant responses remain restricted to a minority of patients. Here, we show a therapeutic strategy that combines enhancing the phagocytic activity of antigen-presenting cells with immunogenic cell death to trigger efficient antitumour immunity. Rho-kinase (ROCK) blockade increases cancer cell phagocytosis and induces antitumour immunity through enhancement of T-cell priming by dendritic cells (DCs), leading to suppression of tumour growth in syngeneic tumour models. Combining ROCK blockade with immunogenic chemotherapy leads to increased DC maturation and synergistic CD8+ cytotoxic T-cell priming and infiltration into tumours. This therapeutic strategy effectively suppresses tumour growth and improves overall survival in a genetic MMTV/Neu tumour model. Collectively, these results suggest that boosting intrinsic cancer immunity using immunogenic killing and enhanced phagocytosis is a promising therapeutic strategy for cancer immunotherapy.

Citation Format: In-San Kim, Gi Beom Kim, Gi-Hoon Nam, Yeonsun Hong. Combined Rho-kinase inhibition and immunogenic cell death triggers and propagates immunity against cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3966.
Abstract 3991: ONCOTARGET (ONCOALVO): A custom NGS panel for therapeutic decision in solid tumors refractory to conventional therapy
Authors: Murad Andre M. , Casali-da-Rocha Jose Claudio , Carneiro Juliana G. , Godard Ana L.
Published: August 2019

Next-generation sequencing (NGS) of tumor biopsies of both solid tumors, as well as hematological malignancies, using commercially available platforms has broadly entered routine clinical practice in medical oncology. This molecular diagnostic approach is now used mainly to test for predictive biomarkers for patients with no available further standard therapies, as diagnostics rely on genomic testing of molecular alterations to enable effective cancer treatment. However, the high cost of large multigenic panels and especially the sequencing of the entire tumor exome is a concern, especially in a developing country such as Brazil. Here we report the clinical application of a panel of 57 genes that we call ONCOTARGET (ONCOALVO) which is based on the Thermo Fisher Oncomine Focus Assay, an integrated, commercially available NGS assay for the rapid and simultaneous detection of single nucleotide variants, short insertions and deletions, copy number variations, and gene rearrangements in 52 cancer genes with therapeutic relevance, but with 5 additional genes: the homologous recombination repair (HRR) genes BRCA1, BRCA2, ATM and CHEK2 that determine tumor sensitivity to inhibitors of the PARP enzyme and also KDR, which determines sensitivity to regorafenib. Twenty-five diagnostic samples of formalin-fixed, paraffin-embedded material submitted for molecular testing over a 8-month time period were analyzed so far. All patients had advanced solid tumors already refractory to conventional systemic therapy. Libraries were prepared from isolated nucleic acids and sequenced on the Ion Torrent S5 sequencer. Sequencing datasets were analyzed using the Ion Reporter software. From the samples of the 39 patients tested, we found 30 potential therapy target pathogenic gene aberrations in 22 (56.4%) patients: KRAS - 7 (23.3%), BRCA1 - 6 (20%), BRCA2 - 5 (16.6%), PIK3CA - 3 (10%), MAP2K1 - 2 (6.6%), BRAF-V600E - 2 (6.6%) mutations; and also one of each: ERBB2 (3.3%), mTOR (3.3%), KIT (3.3%), ALK (3.3%) mutations and one (3,3%) CCND1 amplification. In 3 (7,69%) patients there were double mutations: BRCA1 and BRCA2 in 2 cases and BRCA2 and KRAS and one case. The Oncotarget workflow enabled a turnaround of 18 frac12 days. Taken together, ONCOTARGET was found to be a convenient tool for fast, reliable, broadly applicable and cost-effective targeted NGS of tumor samples in routine diagnostics and therapeutic decisions with a potential to become an important assay for precision oncology in Brazil.

Citation Format: Andre M. Murad, Jose Claudio Casali-da-Rocha, Juliana G. Carneiro, Ana L. Godard. ONCOTARGET (ONCOALVO): A custom NGS panel for therapeutic decision in solid tumors refractory to conventional therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3991.
Abstract 4: Discovery of novel and highly potent small molecule inhibitors of chemokine receptor CXCR4
Authors: Fang Xiong , Meng Qian , Fang Xiao , Zhu Siyu , Huang Yazi , An Jing , Xu Yan , Huang Ziwei
Published: August 2019

Introduction: CXCR4 is a member of chemokine receptor and G protein coupled receptor (GPCR) families. Its interaction with the chemotactic ligand stromal cell-derived factor 1 (SDF-1 or CXCL12) plays important roles in physiological and pathological processes. Recent studies have showed that the blockade of CXCL12-CXCR4 interaction by small molecules can be used in several clinical applications, including sensitizing cancer cells to chemotherapy, mobilizing hematopoietic stem cells (HSCs) to the blood for HSC collection, and inhibiting the tumor metastasis. To date, AMD3100 (Plerixafor) is the only clinically approved drug used in combination with G-CSF for mobilizing HSCs. Here, we report the discovery and development of a new class of highly potent small molecule CXCR4 inhibitors which can strongly antagonize CXCL12-CXCR4 interaction and mobilizing HSCs. Methods: Based on the crystal structures of CXCR4 showing a major and minor subpockets, we applied a fragment-based combinatorial design (FBCD) strategy to design a new class of hybrid molecules to recognize these two subpockets. A series of designed compounds were chemically synthesized and tested in a panel of biological assays including competitive CXCR4 binding, chemotaxis, calcium mobilization and CXCR4 internalization to characterize their biological activities. Furthermore, site-directed mutagenesis coupled with molecular docking simulation was carried out to determine the binding modes of the most potent compounds. Results: More than 30 compounds were synthesized and characterized. Among them, compound HFX51116 showed the most potent CXCR4 binding activity (IC50 = 12.2 nM), which was more potent than AMD3100 tested in parallel (IC50 = 325.3 nM). Furthermore, HFX51116 displayed remarkable inhibition of calcium mobilization (IC50 = 1.478 nM) and chemotaxis activity (IC50 = 252.7 nM), both of which are better than those of AMD3100. In addition, mutational studies and molecular docking simulation suggested that HFX51116 recognizes the minor subpocket mainly and major subpocket partially. Conclusions: Our findings have demonstrated that HFX51116 is a promising novel lead compound for developing a new class of small molecule therapeutics targeting CXCR4.

Citation Format: Xiong Fang, Qian Meng, Xiao Fang, Siyu Zhu, Yazi Huang, Jing An, Yan Xu, Ziwei Huang. Discovery of novel and highly potent small molecule inhibitors of chemokine receptor CXCR4 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4.
Abstract 4005: Glycemic microenvironment and ovarian cancer risk in underserved minority populations
Authors: Henry Desmond , Washington Crystal , Rashad Marley , Fan Guihong , Rao Veena , Frazier Monica
Published: August 2019

Studies have shown that ovarian cancer risk is associated with diabetes mellitus (DM). Moreover, the notion that cancer growth and progression is linked to obesity is also widely accepted. However, the impact of the glycemic and adipose microenvironment is often overlooked. Epidemiologic studies of these microenvironments in medically underserved populations are also overlooked. The aim of this study was to determine the relationship between DM and ovarian cancer incidence and progression in underserved minority populations.

A retrospective study of ovarian cancer patients diagnosed over at 10 year period (2008 – 2018) at one location was performed. Data abstracted for this study included demographic, pathologic, glycemic, DM and BMI data of 142 women with ovarian cancer. Pearson chi-square (p<0.05) was used to compare variables.

26 (18%) of 141 patients had a DM diagnosis and 128 (91%) of the 141 were from an underserved minority group. There was an inverse relationship between tumor size and stage. Patients in later stages had significantly smaller tumors than those in earlier stages of ovarian cancer. There was also a direct correlation between hemoglobin a1c (HbA1c) levels and tumor staging in patients with more advanced disease having slightly higher HbA1c levels (5.2, 5.5, 5.3 and 5.8 HbA1c levels Stages 1 - 4, respectively) And although there continues to be discussion on the best glycemic indicator for determining risk, we did find a similar correlation between staging and blood glucose levels. Lastly, when investigating the relationship between BMI and ovarian cancer staging, it appears that stage 1 occurrence was irrespective of BMI measurements (range 16.3 – 63.2). However, in stages 2 (range 17 – 22.8), 3 (range 21.5 – 33.9) and 4 (range 20.9 – 41.2), there was a positive correlation between BMI and staging. This result suggests that ovarian cancer progression is associated with BMI.

Overall, when using cancer stage as an indicator of ovarian cancer progression, this data suggests that while tumor size is inversely related to progression, the glycemic microenvironment along with the increased presence of adipose tissue is positively correlated with ovarian cancer progression.

Citation Format: Desmond Henry, Crystal Washington, Marley Rashad, Guihong Fan, Veena Rao, Monica Frazier. Glycemic microenvironment and ovarian cancer risk in underserved minority populations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4005.
Abstract 4027: XNA-Based OptiSeqTMlung and colorectal cancer mini panel, a high sensitivity method for cancer early detection
Authors: Sha Michael Y. , Feng Wenjing , Sun Qing , Zhan Ke , Powell Michael J. , Zhang Aiguo
Published: August 2019

The identification of genetic variants with low allelic frequency using next-generation sequencing method is confounded by the complexity of human genome sequence and by bias that arise during library preparation, sequencing and analysis. Herein, we introduce a novel molecule, Xenonucleic Acid (XNA) for NGS application. XNA is able to selectively suppress primer directed amplification of DNA with wild type alleles and only amplify DNA target templates containing mutant alleles. Mutants with low allelic frequency will be easily detectable without deep sequencing after enrichment by adding XNA in multiplex PCR. The 17 actionable mutants related to lung or colorectal cancer diseases at different VAF% were investigated in this study. Upon XNA blocking of wild type alleles, enriched variant allelic frequency (VAF) can be increased by ~32 fold from 10 ng of gDNA samples containing mutants as low as 0.10% VAF. Analytical sensitivity of Limit of Detection (LOD) is about 0.10% VAF. These 17 actionable mutants were tested and verified by using tissue biopsy (FFPE) and liquid biopsy (cfDNA) of lung or colon cancer patient samples. Clinical sensitivity for FFPE sample is about 100% for lung cancer and colorectal cancer samples respectively, comparing to without XNA NGS about 85.7% for lung cancer and 70% for colon cancer. For cfDNA sample its clinical sensitivity is about 100% for lung and colon cancer, but without XNA mediated enrichment NGS is only about 70% for lung cancer and undetectable for early colon cancer. This method provides a simple, robust, accurate, faster turnaround, highly sensitive and low cost alternative compared conventional NGS employing deep sequencing. This technology will greatly enhance the adaptability of NGS as a clinically useful platform for every modern pathology laboratory.

Citation Format: Michael Y. Sha, Wenjing Feng, Qing Sun, Ke Zhan, Michael J. Powell, Aiguo Zhang. XNA-Based OptiSeqTMlung and colorectal cancer mini panel, a high sensitivity method for cancer early detection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4027.
Abstract 403: COX-2 expression on circulating tumor cells in nasopharyngeal carcinoma predicts a worse survival: A prospective analysis
Authors: Li Yongjiang , Xie Xiaoqi , Wang Feng
Published: August 2019

COX-2 expression on circulating tumor cells in nasopharyngeal carcinoma predicts a worse survival: A prospective analysis

Yong-Jiang Li1, Xiao-Qi Xie2, Feng Wang1*

1Department of Oncology, West China Hospital, Sichuan University, Chengdu, PR China

2Department of Critical Care Medicine, West China Hospital, Sichuan University, Chengdu, PR China

*Corresponding author

Short title: COX-2 Expression on CTCs in NPC patients

Abstract

Background and purpose: The prognostic significance of circulating tumor cells (CTCs) in head and neck cancer is still under debate, as only a few studies have been reported and limited conclusions are reached. Besides, CTCs count alone was utilized as an indicator in the previous researches. As a form of ‘liquid biopsy’, the further identification of genetic or phenotypic biomarkers on CTCs could possibly provide further clinical significance.

Materials and methods: The prospective study enrolled 131 patients with nasopharyngeal carcinoma (NPC). CTCs were isolated at baseline and at the end of concurrent chemoradiotherapy, and cyclooxygenase-2 (COX-2) expression status of CTCs was detected by RNA-in situ hybridization (ISH) method. Results were correlated with patient’s clinicopathological parameters and treatment outcomes. Univariate and multivariate survival analysis were performed to determine the prognostic significance.

Results: COX-2 expression was found in 87/131 (66.4%) patients at baseline and 53/115 (46.1%) patients at the end of concurrent chemoradiotherapy. Patients with post-therapeutic COX-2 expression had significantly poorer treatment response (P=0.011) and higher risk of tumor relapse (P=0.026) and metastasis (P=0.007). Besides, multivariate analysis revealed that post-therapeutic COX-2 expression on CTCs remained an independent prognostic indicator for poorer

progression-free survival (HR 2.17, P=0.019) and overall survival (HR 2.41, P=0.024).

Conclusion: The study demonstrated that post-therapeutic COX-2 expression on CTCs was a novel and promising prognostic indicator for NPC patients. Future studies are needed to validate our findings and further clarify the value of integrating the indicator with current clinical strategies in improving survival of NPC patients.

Note: This abstract was not presented at the meeting.

Citation Format: Yongjiang Li, Xiaoqi Xie, Feng Wang. COX-2 expression on circulating tumor cells in nasopharyngeal carcinoma predicts a worse survival: A prospective analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 403.
Abstract 4081: Efficacy of MERTK inhibitor in combination with pembrolizumab in lung cancer
Authors: Pyo Kyoung-Ho , Cho Ha-Ni , Xin Chun-Feng , Cho Jae Seok , Kang Han Na , Yun Jiyeon , Kim Hye Ryun , Cho Byoung Chul
Published: August 2019

Introduction: MERTK is thought to play a role in cancer immune evasion mechanism based on M2 type macrophage-driven immunosuppression in tumor microenvironment (TME). Thus, MERTK is a highly promising target for anti-cancer therapy under high tumor associated macrophages (TAMs) condition. We evaluated the anti-tumor effects and immune reaction of combination or single MERTK inhibitor in two lung squamous patients derived xenograft (PDX) tumor engrafted Hu-CD34-NSG models.

Methods: The aPD-1 sensitive (YHIM2004) and aPD-1 non-sensitive (YHIM2009) squamous cell carcinoma PDX models were selected from pre-test with pembrolizumab. The selected two models were composed four groups; control, pembrolizumab, MERTK inhibitor, and combination. We applied pembrolizumab as aPD-1 blockade (10 mpk, q2w, i.p. inj.). The dose of MERTK inhibitor was 50 mpk, q1d by orally. The treatment was started when tumor size reached 100 mm3. Whole exome sequencing (WES), microarray (mRNA), flow cytometry, and multispectral image analysis were performed on the tumors. To elucidate the involvement of MERTK inhibitor in tumor, the tumor mutation burden, tumor microenvironment and immune cell phenotype were deconvoluted.

Results: The TMB of YHIM2004 and YHIM2009 was 3.23 and 2.64 (mutations/Mb) respectively. The PD-L1 and TIL level of YHIM2004 were relatively higher than YHIM2009. Both two models were observed high macrophage in tumor. The two PDX-models, YHIM2004 (aPD-1 sensitive), YHIM2009 (aPD-1 non-sensitive) were observed anti-tumor effects in MERTK inhibitor single treatment groups (TGI = 44.23%, 67.40% respectively, p < 0.05, t-test). Especially, YHIM2004 tumor was reduced after combination therapy more than single treatment. Of note, the NOG-PDX model demonstrated no anti-tumor effect in MERTK inhibitor monotherapy, suggesting that this drug involves anti-cancer immune responses. Flow cytometry data showed increased PD-1+ and granzyme B+ T cells (p <0.05, t-test) in the combined group (YHIM2004) and MERTK inhibitor monotherapy group (YHIM2009) as much as the tumor suppressive effect.

Conclusion: We concluded that MERTK signaling involves regulation of anti-cancer immune responses. Our results suggest an anti-cancer therapeutics of MERTK selective inhibitor on aPD-1 sensitive or non-sensitive tumors under high TAM tumor microenvironment.

Citation Format: Kyoung-Ho Pyo, Ha-Ni Cho, Chun-Feng Xin, Jae Seok Cho, Han Na Kang, Jiyeon Yun, Hye Ryun Kim, Byoung Chul Cho. Efficacy of MERTK inhibitor in combination with pembrolizumab in lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 4081.