bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of Acupuncture on Muscle Endurance in the Female Shoulder Joint: A Pilot Study
Authors: Wang I.-Lin , Chen Yi-Ming , Hu Rui , Wang Jun , Li Zheng-Bin
Published: September 2020

Shoulder joint dysfunction is the leading cause of decreased athletic ability in athletes. Shoulder joint sports injuries affect the athletic performance of athletes. Improvements in the muscle endurance of the shoulder joint can reduce the incidence of shoulder joint dysfunction. Acupuncture has been an important part of Asian culture for a long time. In acupuncture, nerves are stimulated, inducing postactivation potentiation (PAP) in the body’s motor units and enhancing muscle strength. In this research, 20 female participants with full flexion/extension and adduction/abduction ranges of motion in the shoulder joint during isokinetic exercises underwent stimulation of the following acupuncture points in the shoulder joint: Binao (LI14), Jianyu (LI15), Jianliao (SJ14), Naohui (SJ13), Yuzhong (KI26), Zhongfu (LU1), Yunmen (LU2), Xiabai (LU4), Chize (LU5), Tianfu (LU3), and Xiaoluo (SJ12). In the study, there were significant increases after acupuncture in the average maximum torque in flexion, extension, and adduction; the average work in flexion/extension and adduction/abduction; the average power in flexion/extension and adduction/abduction; the total work in flexion/extension and adduction/abduction; the total net sagittal‐plane work (flexion + extension); and the total net frontal‐plane work (adduction + abduction) (P < 0.05). The average maximum abduction torque did not increase significantly, potentially due to antagonistic forces of muscles. Therefore, acupuncture at acupoints around the shoulder joint can increase muscle excitability, thereby delaying muscle fatigue and increasing muscle endurance.
Abstract 1007: Orphan nuclear receptor TLX regulates the antitumor immune response through transcriptional regulation of programmed death ligand 1 in glioma
Authors: Zhou Jiayi , Pei Xiaojuan , Wang Zhu , Gao Weijie , Chan Franky Leung , Li Xin , Mao Jie , Wu Dinglan
Published: September 2020

High-grade gliomas are rapidly progressing tumors of the central nervous system (CNS) with a very poor prognosis and novel treatment are urgently needed to improve patient outcomes. Clinical trials of immunotherapies for glioma including PD1/PD-L1 inhibition have performed but the outcome was not optimistic. In this study, we found that orphan nuclear recepter TLX and PD-L1 were both significantly upregulated in glioma lesions and the expression of TLX and PD-L1 in glioma was positively correlated. Moreover, the expression pattern of TLX also associated with a suppression microenvironment in glioma. Functional study revealed that suppression of TLX inhibited glioma cell growth and immune response, which accompanied with down-regulation of PD-L1. We also characterized that TLX could bind directly to PD-L1 promoter and activate PD-L1 gene transcription. Together, our present study shows, for the first time, that TLX can contribute to glioma immune escape via promoting the expression of PD-L1, targeting the druggable TLX may have a potential therapeutic significance in glioma immune therapy.

Citation Format: Jiayi Zhou, Xiaojuan Pei, Zhu Wang, Weijie Gao, Franky Leung Chan, Xin Li, Jie Mao, Dinglan Wu. Orphan nuclear receptor TLX regulates the antitumor immune response through transcriptional regulation of programmed death ligand 1 in glioma [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 1007.
Abstract 1059: Targeting IDH1/2 mutant cancers with unique combinations of DNA repair inhibitors
Authors: Sule Amrita D. , Sundaram Ranjini , Bindra Ranjit
Published: September 2020

Mutations in the Isocitrate Dehydrogenase-1 and -2 (IDH1/2) genes occur in the vast majority of solid tumors including low-grade and secondary high-grade gliomas. These neomorphic mutations occur early on in gliomagenesis leading to the production of 2-Hydroxyglutarate (2HG). 2HG has been implicated in tumorigenesis via inhibiting α-ketoglutarate (αKG)-dependent dioxygenases. Our group recently demonstrated that the production of 2HG suppresses the high-fidelity homologous recombination (HR) DNA repair pathway, resulting in a state of “BRCAness”. We initially found that mutant IDH1/2-induced BRCAness confers exquisite sensitivity to poly(ADP-ribose) polymerase (PARP) inhibitors, a finding which now has been replicated by multiple independent laboratories. Although IDH1/2 mutations were first identified in gliomas and acute myeloid leukemia (AML) cells, multiple other tumor types have subsequently been shown to harbor these mutations. Current clinical trials are testing the efficacy of PARP inhibitors as a monotherapy, as well as in combination with other DNA repair inhibitors. Here, we demonstrate that novel combinations of DNA repair inhibitors can be utilized to synergistically target IDH1/2-mutant glioma cells. In particular, we demonstrate potent synergy with ATRi and PARPis, a finding which was validated in multiple structurally unique drugs within these classes. We have shown that in an in vivo flank model bearing IDH1 R132H mutation, AZD6738 (ATRi) and Olaparib (PARPi) combination treatment reduces the tumor burden significantly relative to monotherapy and vehicle. As this combination is active in BRCA1/2-mutant cancers, in particular after the emergence of PARPi resistance, these data suggest are consistent with an underlying HR defect in IDH1/2-mutant gliomas. These preclinical investigations will provide a blueprint for future clinical trials combining PARP and ATR inhibitors in the treatment of IDH1/2 mutant cancers.

Citation Format: Amrita D. Sule, Ranjini Sundaram, Ranjit Bindra. Targeting IDH1/2 mutant cancers with unique combinations of DNA repair inhibitors [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 1059.
Abstract 1074: Programmable bacteria induce durable tumor regression and systemic antitumor immunity
Authors: Chowdhury Sreyan , Pu Kelly , Nimura Clare , Castro Samuel , Coker Courtney , Hinchliffe Taylor , Arpaia Nicholas , Danino Tal
Published: September 2020

Synthetic biology is driving a new era of medicine through the engineering of living cells. This transformative approach allows for the creation of systems that intelligently sense and respond to diverse environments, adding specificity and efficacy beyond the capabilities of molecular-based therapeutics. One particular area of focus has been the engineering of bacteria as therapeutic delivery systems to selectively release therapeutic payloads in vivo. We recently engineered a non-pathogenic Escherichia coli strain to specifically lyse within the tumor microenvironment and release an encoded nanobody antagonist of CD47 (CD47nb), an anti-phagocytic receptor that is commonly overexpressed in several human cancer types. We show that delivery of CD47nb by tumor-colonizing bacteria increases activation of tumor-infiltrating T cells, stimulates rapid tumor regression, prevents metastasis and leads to long-term survival in a syngeneic tumor model in mice. Moreover, we report that local injection of CD47nb-expressing bacteria stimulates systemic tumor-antigen-specific immune responses that reduce the growth of untreated tumors, providing proof-of-concept for an abscopal effect induced by an engineered bacterial immunotherapy. We have expanded this approach to include additional human and murine specific immunotherapeutic payloads and evaluate efficacy in multiple tumor model systems.

Citation Format: Sreyan Chowdhury, Kelly Pu, Clare Nimura, Samuel Castro, Courtney Coker, Taylor Hinchliffe, Nicholas Arpaia, Tal Danino. Programmable bacteria induce durable tumor regression and systemic antitumor immunity [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 1074.
Abstract 1079: Distinct DNA methylation targets by aging and chronic inflammation
Authors: Yamashita Satoshi , Nanjo Sohachi , Rehnberg Emil , Iida Naoko , Takeshima Hideyuki , Ando Takayuki , Maekita Takao , Sugiyama Toshiro , Ushijima Toshikazu
Published: September 2020

Aberrant DNA methylation is involved in many types of human cancers, and is induced by aging. In addition, it is known to be induced by chronic inflammation, and this inflammation-induced DNA methylation is considered as acceleration of age-related methylation. However, few studies addressed target genomic regions of age-related and inflammation-induced DNA methylation in a genome-wide manner. We previously demonstrated that Helicobacter pylori(HP) infection induces aberrant DNA methylation potently in human and gerbil gastric mucosa [Niwa, Cancer Res, 70:1430, 2010; Ushijima, Clin Cancer Res, 18:923, 2012]. Here, taking advantage of the potent methylation induction, we aimed to clarify the targets of methylation induction by aging and inflammation. We collected gastric mucosae from i) HP-never infected young individuals (under 40 years; n=4; G1), ii) HP-currently infected young individuals (n=4; G2), iii) HP-never-infected old individuals (above 60 years; n=4; G3), and vi) HP-currently-infected old individuals (n=4; G4). DNA methylation of 482,421 CpG probes was analyzed by Infinium Human Methylation 450K, and the probes were grouped into 270,249 genomic blocks. It was revealed that high levels of methylation were induced in 44,461 (16.5%) genomic blocks by current inflammation (G2/G1), even after correction of the influence of leucocyte infiltration. 61.8 % of the hypermethylation was acceleration of age-related methylation (G3/G1) while 21.6 % was specific to inflammation. Regions with H3K27me3 were frequently hypermethylated both by aging and inflammation. Basal methylation levels were essential for age-related hypermethylation while even regions with little basal methylation were hypermethylated by inflammation. When limited to promoter CpG islands, being a microRNA gene and high basal methylation levels strongly enhanced hypermethylation while H3K27me3 strongly enhanced inflammation-induced hypermethylation. Inflammation was capable of overriding active transcription. In young gastric mucosae (G2/G1), genes with high expression and frequent mutations in gastric cancers were more frequently methylated than in old ones (G4/G3).Methylation by inflammation was not simple acceleration of age-related methylation. Targets of aberrant DNA methylation were different between young and old gastric mucosae, and driver genes were preferentially methylated in young gastric mucosa [Yamashita, Clin Epigenet, in press].

Citation Format: Satoshi Yamashita, Sohachi Nanjo, Emil Rehnberg, Naoko Iida, Hideyuki Takeshima, Takayuki Ando, Takao Maekita, Toshiro Sugiyama, Toshikazu Ushijima. Distinct DNA methylation targets by aging and chronic inflammation [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 1079.
Abstract 1144: Novel prostate cancer mouse models for testing targeted chemopreventive potential of pterostilbene
Authors: Hemani Rutu , Inamdar Ninad , Kumar Avinash , Donovan Virginia , Levenson Anait S.
Published: September 2020

Metastasis-associated protein 1 (MTA1) plays role in prostate cancer progression and metastasis. We have previously reported on the MTA1-targeted chemopreventive and therapeutic efficacy of pterostilbene (Pter) in the prostate-specific Pten heterozygous (Pten+/f; Pb-Cre+) and Pten null (Ptenf/f; Pb-Cre+) mice. To investigate the exact role of MTA1 in prostate cancer tumorigenesis and its early stages, we developed appropriate mouse models, namely prostate-specific MTA1-knocked in (MTA1-tg) and MTA1-tg on the background of prostate-specific Pten heterozygosity (MTA+/+, Pten+/f; Pb-Cre+). In the current project, we asked three major questions: 1) whether prostate MTA1 overexpression in the context of Pten loss promotes an earlier onset, greater total incidence and accelerated PIN or tumor growth rate; 2) what MTA1-associated molecules are involved in prostate cancer progression; and 3) whether Pter supplemented diet can provide beneficial effects. Identified from our ChIP-Seq data MTA1-associated molecules included CyclinD1, Notch2, and two neuroendocrine markers, namely CHGA and SYP. Using two prostate cancer cell lines, DU145 and PC3M and their corresponding MTA1 knockdown (shMTA1) clones, we validated the direct link between MTA1 and these markers on mRNA and protein levels. Further, we generated 28 prostate-specific MTA1+/+; Pten+/f; Pb-Cre+ mice and randomized them into two major groups: 1) mice on control diet (Ctrl-Diet) and 2) mice on Pter-Diet. After 5 months of feeding with corresponding diets (AIN-76A phytoestrogen free diets supplemented or not with Pter at 100 mg/kg/diet), mice were sacrificed and prostate tissues were isolated for histological, immunochemical and molecular analysis. Results demonstrated the contribution of MTA1 in more aggressive histological phenotype (Ki67, PCNA, p27, SMA, CK8), increased levels of oncogenic MTA1-guided molecules (CyclinD1, Notch2, CHGA, SYP, AR) and the potential for Pter in prostate cancer chemoprevention.

Citation Format: Rutu Hemani, Ninad Inamdar, Avinash Kumar, Virginia Donovan, Anait S. Levenson. Novel prostate cancer mouse models for testing targeted chemopreventive potential of pterostilbene [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 1144.
Abstract 1146: Development of CRISPR human Skp2 knock-in in the prostate of mouse and the associated prostate organoids for testing Skp2 targeting agents
Authors: Song Liankun , Xu Shan , Arabzadehkaffash Kia , Fazelpour Ali , Fu Dongjun , Tippin Matthew , Zi Xiaolin
Published: September 2020

S-phase kinase associated protein 2 (Skp2) is a promising drug target as studies have demonstrated that Skp2 is required for spontaneous tumor development that occurs in the retinoblastoma protein (pRb), Pten or p53 deficient mice. We therefore have aimed to establish CRISPR human Skp2 (hSkp2) knock-in in the prostate of mouse to examine its role in prostate carcinogenesis. In addition, Skp2 overexpressing organoids are being developed for convenient and efficient screening of specific Skp2 inhibitors. cDNA of hSkp2 and porcine teschovirus-1 (P2A) was introduced into immediate upstream of mouse probasin coding sequencing using CRISPR knock-in targeting approach to establish transgenic mice specifically overexpressing hSkp2 in the prostate glands. Quantitative PCR, western blot and immunohistochemistry (IHC) were used to identify the expression levels of Skp2 and histopathological analyses were performed after H&E staining. Prostate glands were dissected and digested for organoid culture, then morphological features and viability of organoids were evaluated following treatments of potential Skp2 inhibitors. Prostate glands and other tissues from mice of three transgenic lines which were genotyped positive were dissected and hSkp2 was detected in ventral, dorsal and lateral prostate lobes, but not in testis, kidney, spleen, and liver. Line 10092 was found to have the highest expression of hSkp2 among these three lines. Prostate weight slightly increased in transgenic mice compared to that of wild-type control mice. Hyperplasia, prostatic intraepithelial neoplasia (PIN), and carcinoma were noted, which suggested that overexpression of hSkp2 in the prostate of mice promoted proliferation and prostate tumorigenesis. Organoids that were derived from the prostate of hSkp2 transgenic mice recapitulated prostatic phenotypic features and maintained a high level of hSkp2. A novel Flavokawain A derivative was found to selectively alter the morphology of hSkp2 organoids and decrease the viability. Prostatic lesions found in the hSkp2 knock-in mice demonstrated a potential role of hSkp2 in early prostate carcinogenesis and organoids overexpressing hSkp2 have the utility for screening out compounds targeting Skp2 in prostate cancer.

Citation Format: Liankun Song, Shan Xu, Kia Arabzadehkaffash, Ali Fazelpour, Dongjun Fu, Matthew Tippin, Xiaolin Zi. Development of CRISPR human Skp2 knock-in in the prostate of mouse and the associated prostate organoids for testing Skp2 targeting agents [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 1146.
Abstract 115: Hedgehog signaling pathway promotes breast cancer adaptation to hypoxia
Authors: Lama-Sherpa Tshering D. , Kammerud Sarah C. , Shevde Lalita A.
Published: September 2020

Breast cancer is the most commonly diagnosed cancer in women in the US. Metastasis accounts for the majority of breast cancer-related deaths. Rapidly proliferating primary tumors experience oxygen levels that are lower in comparison to the normal tissue. The proportions of these hypoxic tumor cells in breast cancer strongly correlate to the incidence of metastasis. Around 70% of breast cancers present with bone metastasis. Tumor cells that metastasize to the bone can lead to an imbalance in bone homeostasis, often leading to osteolysis. Our lab has previously reported that the activation of Hedgehog (Hh) signaling, one of the embryonic developmental pathways found to be abnormally activated in breast cancer, leads to increased osteolysis in vivo. The incidence of osteolytic metastasis at the tibio-femoral junction was significantly reduced in the Hh signaling-deficient mice. Since a hypoxic microenvironment is prevalent in bone, it is imperative to understand how Hh signaling drives tumor cell adaptation to hypoxia. The hypoxic tumor microenvironment in the primary tumor might select for metastatic cells that are better able to metastasize to the bone. We hypothesize that Hh signaling enables the adaptation of breast cancer cells to hypoxia and consequently their metastatic potential. Hh signaling can be canonically activated through smoothened (SMO)-dependent activation of the transcriptional regulator GLI. Cellular response to hypoxia is mediated by the hypoxia-inducible factors (HIFs). To understand the importance of Hh signaling activation in hypoxic condition, we inhibited the Hh signaling pathway using GLI and SMO inhibitors. Additionally, we knocked-down GLI1 in breast cancer cell lines. We have identified a feed-forward mechanism between Hh signaling and HIF signaling in hypoxic conditions leading to sustained hypoxia response by stabilization of HIF-1α. Future investigations will address the significance of Hh signaling inhibition in a murine model of TNBC. This study will help determine the therapeutic benefit of hedgehog inhibitors in controlling hypoxia-mediated metastasis and tumorigenesis.

Citation Format: Tshering D. Lama-Sherpa, Sarah C. Kammerud, Lalita A. Shevde. Hedgehog signaling pathway promotes breast cancer adaptation to hypoxia [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 115.
Abstract 1239: Role of the glutathione antioxidant system in the autophagic process outcome
Authors: Lee Yong J.
Published: September 2020

Ferroptosis has been reported as a unique form of cell death. However, in recent years, researchers have increasingly challenged the uniqueness of ferroptosis compared to other types of cell death. In this study, we examined whether ferroptosis shares cell death pathways with other types of cell death, especially autophagy, via the autophagic process. Here, we observed that ferroptosis inducers (artesunate and erastin) and autophagy inducers (bortezomib and XIE62-1004) led to autophagosome formation via the endoplasmic reticulum (ER) stress response. Unlike XIE62-1004, artesunate, erastin, and bortezomib, which affect glutathione production or utilization, induced oxidative stress responses - an increase in the levels of heme oxygenase-1 and lipid peroxidation. Oxidative stress responses were attenuated by deletion of autophagy related gene-5 or treatment with autophagy inhibitors (bafilomycin and chloroquine). Our studies provide an overview of common death pathways - the ER stress response-associated autophagic process in ferroptosis and autophagy. We also highlight the role the glutathione redox system plays in the outcome of the autophagic process.

Citation Format: Yong J. Lee. Role of the glutathione antioxidant system in the autophagic process outcome [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 1239.
Abstract 1247: NOX4 P62 regulates tuberculous fibrosis promoted tumorigenic potential
Authors: Hong Yoonki , Woo Seong-Ji , Kim Youngmi , Hong Ji Young
Published: September 2020

Background Increased incidence of lung cancer in subjects with previous tuberculous infection was reported in epidemiologic data. NOX4 signaling contributes to tuberculous pleural fibrosis and lung cancer proliferation, respectively. The objective of this study was to investigate the role of NOX4 in tuberculous pleurisy-assisted tumorigenicity both in vitro and in vivo.

Methods After pleural mesothelial cells from WT and NOX4 KO C57BL/6 mice were stimulated with Heat-Killed mycobacterium tuberculosis (HKMT) for 24 hour, cellular interactions between mesothelial cell and cancer cell were evaluated by using transwell invasion assay and wound healing assay. A murine model injected with cancer at 4 weeks after Mycobacterium bovis bacillus Calmette-Guérin (BCG) pleural infection were used to validate the contribution of tuberculous fibrosis to tumor invasion.

Results HKMT -stimulated mesothelial cells induced enhanced invasion and migration potential of lung cancer cells in a NOX4-dependent manner. The inhibition of NOX4 signaling exerted a negative effect on the metastatic potential of lung cancer by tuberculous fibrosis in mice model. The expression of inflammatory cytokines like TNF-a, TGF-β and IL-6 was enhanced in the WT-BCG+CA than WT+PBS+CA and NOX4 KO+BCG+CA. The expression of P62 in lung tissue was higher in the WT-BCG+CA than WT+PBS+CA and NOX4 KO+BCG+CA.

Discussions Our results suggest that NOX4 and autophagy signaling changed by tuberculous fibrosis could result enhanced tumorigenic potential. NOX4-P62 might seve as a target for the development of anti-tuberculous fibrosis and anti-cancer drugs.

Citation Format: Yoonki Hong, Seong-Ji Woo, Youngmi Kim, Ji Young Hong. NOX4 P62 regulates tuberculous fibrosis promoted tumorigenic potential [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 1247.
Abstract 1521: New roles of ATP in metastasis: Extracellular and macropinocytosis internalized ATP work together to induces epithelial to mesenchymal transition and other early steps of metastasis
Authors: Cao Yanyang , Wang Xuan , Evers Maria , Li Yunsheng , Zhang Haiyun , Chen Xiaozhuo
Published: September 2020

Metastasis is estimated to be responsible for more than 90% of all cancer-related deaths. Metastasis is initiated by induction of epithelial-mesenchymal transition (EMT) of cancer cells, but factors and mechanisms involved in EMT are far from being fully elucidated. Intratumoral extracellular ATP (eATP), at levels of 100-700 μM or 103 to 104 times higher than in normal tissues, has been known to induce epithelial-mesenchymal-transition (EMT) of cancer cells via purinergic receptor signaling. However, the exact induction mechanisms are far from fully known. We previously described that eATP is internalized by cancer cells in vitro and in vivo by macropinocytosis in human non-small cell lung cancer cells, drastically elevates intracellular ATP levels, enhances cell proliferation and resistance to anticancer drugs. In this study, we tested the hypothesis that eATP and macropinocytosis-internalized eATP also induces EMT and other early steps of metastasis. Here we report that eATP, at the concentrations reported in tumors, potently induces expression of matrix metallopeptidases (MMPs), detachment, EMT, migration, and invasion of lung cancer cells. The induction was independent of TGF-β and semi-independent of P2X7 activation. eATP performs these functions not only extracellularly, but also intracellularly after being macropinocytically internalized to further enhance P2X7-mediated EMT, filopodia formation and other early steps of metastasis. The knockout of the macropinocytosis-associated SNX5 gene significantly reduces macropinocytosis and slows down tumor growth in nude mice. Collectively, these results show that eATP functions on these processes not only from outside of cancer cells but also inside after being macropinocytotically internalized. These findings which shed light on eATP's initiator and effector roles in almost every step in early metastasis call for rethinking and rebalancing energy equations of intracellular biochemical reactions and the Warburg effect, and identify eATP and macropinocytosis as novel targets for potentially slowing down EMT, preventing metastasis, and reducing metastasis-related death in cancer patients.

Citation Format: Yanyang Cao, Xuan Wang, Maria Evers, Yunsheng Li, Haiyun Zhang, Xiaozhuo Chen. New roles of ATP in metastasis: Extracellular and macropinocytosis internalized ATP work together to induces epithelial to mesenchymal transition and other early steps of metastasis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1521.
Abstract 1699: IMMUcan: Broad cellular and molecular profiling of the human tumor microenvironment
Authors: Hong Henoch S. , Liechti Robin , Reinhard Christoph , Morfouace Marie M. ,   IMMUcan consortium
Published: September 2020

The development of next generation therapeutics for cancer patients requires an improved understanding of the underlying mechanisms of resistance to existing therapies as well as novel biomarkers that can help identify patients most likely to benefit from the treatments. Most studies addressing these questions have predominantly relied on a limited set of tumor profiling techniques or have been hampered by relatively small patient numbers. In response to this need, the IMMUcan consortium has been formed. Co-led by the EORTC (Brussels, Belgium), SIB (Lausanne, Switzerland), Eli Lilly (Indianapolis, IN, USA) and Merck KGaA (Darmstadt, Germany), IMMUcan is an EU-funded, public-private partnership of expert immunologists, cell biologists, technology developers and clinical key opinion leaders. The mission of IMMUcan is to generate broad molecular and cellular profiling data of the tumor microenvironment from up to 3,000 patients derived from five indications: non-small cell lung cancer, head and neck squamous cell carcinoma, renal cell carcinoma, colorectal cancer and breast cancer. Most of these patients will be treated under standard of care and recruited via the EORTC-SPECTA infrastructure. All acquired tumor samples will undergo detailed and standardized analyses applying diverse profiling modalities including whole exome sequencing to determine the mutational landscape, total RNA sequencing for transcriptome analysis, multiplex immunofluorescence (IF) and imaging mass cytometry (IMC) facilitating the assessment of approx. 80 proteins at single cell resolution. To avoid inter-lab variations, each profiling method (sequencing, IF or IMC) will be performed by a single expert laboratory, which will process all IMMUcan samples. These data will be integrated with clinical information and centralized in a Knowledge Management System. Once published, the entire IMMUcan dataset will be made available to the scientific community for further study and interrogation. IMMUcan also acquires blood samples, including PBMCs, saliva as well as fresh-frozen stool samples. In addition, we have established workflows to generate reports documenting clinically actionable molecular alterations, which are returned to the treating physicians within 6-8 weeks. Thus far, 31 patients have been recruited and first tumor profiling data from these patients will be presented. Funding: IMI2 JU grant agreement 821558, supported by EU's Horizon 2020 and EFPIA.

Citation Format: Henoch S. Hong, Robin Liechti, Christoph Reinhard, Marie M. Morfouace, IMMUcan consortium. IMMUcan: Broad cellular and molecular profiling of the human tumor 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 1699.
Abstract 1763: NEO2734, a novel dual bromodomain and histone acetyltransferase inhibitor, in the treatment of colorectal cancer
Authors: Kuang Chaoyuan , Cai Manbo , Risnik Denise , Giles Francis , Zhang Lin
Published: September 2020

Background: Oncogenic c-MYC activation is seen in a broad spectrum of cancers and is often associated with more aggressive disease and/or poor outcomes. c-MYC expression is often driven by transcriptional super-enhancers which depend on histone modifications to function. We report the results of our studies on the novel suppressor of histone reading and writing, NEO2734, and the effects on colorectal cancer cells.

Methods: Commercially and publicly available colorectal cancer cell lines were used under standard culture conditions. All small molecule treatments were performed with DMSO as the primary solvent and dilutions made in aqueous solution. Commonly employed techniques such as MTS viability assays, immunofluorescent 2-D microscopy, siRNA knockdown, western blotting, and single-color flow cytometry were used.

Results: NEO2734, a dual BET/bromodomain inhibitor and p300/CBP inhibitor, reduced cell growth more than the BET inhibitor OTX-015 and the p300/CBP inhibitor CPI-637. NEO2734 potently induced cell death and markers of apoptosis such as PUMA and TRAIL2/DR5. Inhibition of the caspase by zVAD-fmk eliminated the induction of cell death by NEO2734. Knockdown or knockout of the death proteins PUMA or DR5 additively reduced the amount of NEO2734-induced apoptosis. NEO2734 reduced the expression of downstream targets c-MYC, acetylated histone H3, and acetylated histone-H4. Strikingly, NEO2734 significantly upregulated the ER stress markers BiP and ATF3. NEO2734 also activated immunogenic cell death in vitro, as measured by HMGB1 release and membrane localization of calreticulin. Finally, examination of a panel of cell lines revealed SPOP mutant colorectal cancer cells to be exquisitely sensitive to growth inhibition by NEO2734.

Conclusions: NEO2734 causes growth inhibition and apoptosis of colorectal cancer cells. NEO2734 is more potent than single-target BET inhibitors. Anticipated targets such as c-MYC and acetylated histone are confirmed. NEO2734 appears to trigger cell death through both PUMA and TRAIL mediated apoptosis. NEO2734 also activates ER stress and immunogenic cell death. Work is ongoing to confirm the mechanism of apoptosis and test rational drug combinations such as with chemotherapy and immunotherapy.

Citation Format: Chaoyuan Kuang, Manbo Cai, Denise Risnik, Francis Giles, Lin Zhang. NEO2734, a novel dual bromodomain and histone acetyltransferase inhibitor, in the treatment of colorectal 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 1763.
Abstract 2204: Enhancing anti-tumor efficacy of MART-1-specific TCR-T cells by CRISPR-Cas9-based PD-1 disruption
Authors: Gao Qianqian , Ding Renpeng , Chen Huanyi , Xu Qumiao , Zhu Linnan , Dong Xuan , Gu Ying , Chao Cheng-chi
Published: September 2020

T cell function can be compromised when continuously exposed to tumor antigens by immune checkpoint signaling. Immunotherapies with T cells and inhibitors of repressive checkpoint molecules such as PD-1, CTLA-4, etc. have been shown promising results in cancer clinical trials. Inhibitors of checkpoint molecules have been solely used to block or suppress the corresponding pathways to improve the functionality of T cells and enhance the anti-tumor immunity. However, few reports combined disruption of repressive checkpoint molecules with TCR-T cell therapy for the treatment of cancer patients. In our study, new TCR sequence was attained from T cells stimulated by Melan-A (aa27-35) peptide, but the cytotoxicity of MART-1-specific TCR-T cells was found to be inhibited by expression of PD-L1 in target cells. To overcome this PD(L)-1-mediated suppression, Cas9 mRNA and gRNA were electroporated into primary T cells infected by MART-1 TCR lentivirus. CRISPR/Cas9-mediated PD-1 disruption in MART-1 TCR-T cells recovered T cell cytotoxicity inhibited by PD-L1 overexpression in target cells. In addition, secretion of IFNγ and IL-2 was also increased in PD-1-damaged TCR-T cells. Furthermore, the differences between wild-type and PD-1-disrupted TCR-T cells were characterized by single-cell RNA (scRNA) sequencing. In summary, our study illustrates enhanced therapeutic efficacy and different features of PD-1-eliminated TCR-T cells compared to the control, providing an alternative strategy for current cell therapy.

Citation Format: Qianqian Gao, Renpeng Ding, Huanyi Chen, Qumiao Xu, Linnan Zhu, Xuan Dong, Ying Gu, Cheng-chi Chao. Enhancing anti-tumor efficacy of MART-1-specific TCR-T cells by CRISPR-Cas9-based PD-1 disruption [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 2204.
Abstract 2468: Phosphorylation of Thr55 mediates the tumor suppression activities of p53
Authors: He Fan , Chen Lihong , Chen Jiandong
Published: September 2020

P53 is a transcription factor that plays a central role in the regulation of cell cycle, apoptosis and senescence. In unstressed cells, p53 is kept at low level under the strict control of its negative regulators. Upon DNA damage or oncogenic stress, p53 is activated via post-translational modifications. The phosphorylation of serine and threonine, such as Ser15 and Ser46, play important roles in p53 regulation. However, data for Thr55 (T55) phosphorylation is limited and functions of T55 phosphorylation remain elusive. We have previously defined an intramolecular interaction in p53 which modulates DNA binding. The observation of altered intramolecular interactions in T55A and T55D mutants leads to the hypothesis that T55 phosphorylation might affect p53 functions. We generate a T55 phospho-specific antibody to investigate the regulation through this site. The results show T55 phosphorylation is induced by DNA damage compounds. We induce the expression of wild type and T55A mutant p53 in p53 null H1299 cells via the transduction of doxycycline-inducible lentiviral gene expression plasmids. T55A mutant p53 exhibits significantly weaker inhibition of cell viability compared with wild type p53 containing cell in the existence of DNA damaging compounds. Consistently, T55A mutant p53 induces lower mRNA expression level of several important apoptosis related genes and higher MDM2 level in comparison with wild type p53. Overall, our results reveal that phosphorylation of Thr55 is induced by several DNA damaging compounds and alters the expression of p53 target genes, leading to an enhanced tumor suppression.

Citation Format: Fan He, Lihong Chen, Jiandong Chen. Phosphorylation of Thr55 mediates the tumor suppression activities of p53 [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 2468.
Abstract 2469: RCC2 promotes esophageal cancer growth by regulating activity and expression of the Sox2 transcription factor
Authors: Calderon-Aparicio Ali J. , Yamamoto Hiroyuki , de vitto Humberto , Zhang Tianshun , Wang Qiushi , Bode Ann M. , Dong Zigang
Published: September 2020

RCC2 is a protein located in the centrosome, which ensures that cell division proceeds properly. Previous reports show that RCC2 is overexpressed in some cancers and could play a key role in tumor development, but the mechanisms concerning how this occurs are not understood. Further, no evidence exists regarding its role in esophageal cancer. We studied the relevance of RCC2 in esophageal cancer growth and the mechanisms explaining how this process occurs. RCC2 was highly expressed in mouse and human tissues as well as esophageal cancer cell lines, and this overexpression was associated with tumorigenicity. RCC2 promoted cell proliferation, anchorage-independent growth, and migration. These oncogenic effects were accompanied by overexpression of Sox2, an important transcription factor promoting esophageal cancer. RCC2 upregulated and stabilized Sox2 expression by inhibiting proteasome degradation. Likewise, RCC2 increased the transcriptional activity and promoter binding of Sox2. In vivo studies indicated that RCC2 and Sox2 were overexpressed in esophageal tumors compared to normal controls, and this upregulation occurs in the basal cell layer for both proteins. In conditional knockout mice, RCC2 deletion decreased the tumor nodule formation and progression in the esophagus compared with “wild type” mice. PCNA expression, a cell proliferation marker, was also downregulated in RCC2 knockout mice. Overall, our data show for the first time that RCC2 is an important protein for the stabilization and transcriptional activation of Sox2 and further promotion of malignancy in cancer.

Citation Format: Ali J. Calderon-Aparicio, Hiroyuki Yamamoto, Humberto de vitto, Tianshun Zhang, Qiushi Wang, Ann M. Bode, Zigang Dong. RCC2 promotes esophageal cancer growth by regulating activity and expression of the Sox2 transcription factor [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 2469.
Abstract 2504: Modeling molecular development of breast cancer in canine mammary tumors
Authors: Graim Kiley , Gorenshteyn Dmitriy , Robinson David G. , Carriero Nicholas J. , Cahill James , Chakrabarti Rumela , Goldschmidt Michael H. , Durham Amy C. , Funk Julien , Storey John D. , Kristensen Vessela N. , Theesfeld Chandra L. , Sorenmo Karin U. , Troyanskaya Olga G.
Published: September 2020

Malignancy in cancer is a consequence of the progressive accumulation of mutations in a tumor, with profound implications for drug selection and treatment. However, in human studies, inter-patient variability obscures molecular signatures of tumor progression because patients usually present with a single mammary tumor. In contrast, dogs frequently exhibit multiple naturally occurring mammary tumors in the same individual. Moreover, canine mammary tumors (CMTs) and human breast cancer have similar histopathological profiles and clinical presentation. We leverage the CMT model to elucidate genome-wide molecular changes clinically relevant in human breast cancer, focusing on signals underlying tumor development. We develop a robust, generally applicable, computational analysis framework (FREYA) for analysis of CMTs for comparative oncology. Using FREYA, we RNA profile 89 samples from 16 dogs, and demonstrate that CMTs recapitulate human breast cancer subtypes. We then extract molecular profiles of breast cancer progression at three distinct stages (normal, pre-malignant and malignant) and identify signatures of gene expression reflective of tumor progression. Focusing on the transitions to malignancy, we identify transcriptional patterns and biological pathways specific to malignant tumors and distinct from those characterizing pre-malignant tumors or normal tissue. We find that human breast cancer patients whose tumors exhibit strong CMT malignancy signatures have significantly decreased survival, indicative of the importance of the tumor progression processes identified in CMTs to human breast cancer prognosis. Altogether, our comprehensive genomic characterization demonstrates that CMTs are a powerful translational model of breast cancer, providing insights that inform our understanding of tumor development in humans. To catalyze and support similar analyses and use of the CMT model by other biomedical researchers, we publicly share all of our data and provide FREYA, a robust data processing pipeline and statistical analyses framework, at freya.flatironinstitute.org.

Citation Format: Kiley Graim, Dmitriy Gorenshteyn, David G. Robinson, Nicholas J. Carriero, James Cahill, Rumela Chakrabarti, Michael H. Goldschmidt, Amy C. Durham, Julien Funk, John D. Storey, Vessela N. Kristensen, Chandra L. Theesfeld, Karin U. Sorenmo, Olga G. Troyanskaya. Modeling molecular development of breast cancer in canine mammary 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 2504.
Abstract 2509: Genetic heterogeneity and evolutionary history of high grade ovarian carcinoma and matched distant metastases
Authors: Masoodi Tariq , Siraj Sarah , Siraj Abdul K. , Azam Saud , Qadri Zeeshan , Parvathareddy Sandeep K. , Alkuraya Fowzan S. , Al-Kuraya Khawla S.
Published: September 2020

BACKGROUND: High-grade serous ovarian carcinoma (HGSOC) is the most frequent type of ovarian carcinoma, associated with poor clinical outcome and metastatic disease. Although, metastatic processes are becoming more understandable, the genomic landscape and metastatic progression in HGSOC has not been elucidated.

METHODS: Multi-region whole-exome sequencing was performed on HGSOC primary tumors and their metastases (n=33 tumor regions) from 6 patients. The resulting somatic variants were analyzed to delineate tumor evolution and metastatic dissemination, and to compare the repertoire of events between primary HGSOC and metastasis.

RESULTS: All cases presented branching evolution patterns in primary HGSOC, with three cases further showing parallel evolution in which different mutations on separate branches of a phylogenetic tree converge on the same gene. Furthermore, linear metastatic progression was observed in 67% of cases with late dissemination in which the metastatic tumor mostly acquires the same mutational process active in primary tumor, and parallel metastatic progression, with early dissemination in the remaining 33.3% of cases. Metastatic specific SNVs were further confirmed as late dissemination events. We also found the involvement of metastatic-specific driver events in the Wnt/β-catenin pathway.

CONCLUSIONS: This study provides deeper insights into clonal evolution and mutational processes that can pave the way to new therapeutic targets.

Citation Format: Tariq Masoodi, Sarah Siraj, Abdul K. Siraj, Saud Azam, Zeeshan Qadri, Sandeep K. Parvathareddy, Fowzan S. Alkuraya, Khawla S. Al-Kuraya. Genetic heterogeneity and evolutionary history of high grade ovarian carcinoma and matched distant metastases [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 2509.
Abstract 2528: Tumor-derived exosomal miR-619-5p promotes tumor angiogenesis and metastasis through the inhibition of RCAN1.4
Authors: Jung Sang Yong , Kim Dong Ha , Cho Yong-Hee , Choi Yun Jung , Kim Seon Ye , Sung Ki Jung , Lee Chae Won , Lee Hyeon Jeong , Choi Chang-Min , Lee Jae Cheol , Rho Jin Kyung
Published: September 2020

Tumor-derived exsosomes (TEXs) contain enriched miRNAs act as novel non-invasive biomarkers for cancer diagnosis, and they are involved in cancer progression. In this study, we aimed to investigate which and how exosomal miRNA affect cancer progression in non-small cell lung cancer (NSCLC). We found that the only of A549 cell-released exosomes promote an angiogenic phenotype in HUVEC cells compared to other lung cancer cell-released exosomes. Using a miRNA array and RNA sequencing approach, we identified that specific miRNAs can modulate angiogenesis in A549 cell-released exosomes. Among these miRNAs, miR-619-5p significantly induced angiogenesis in HUVEC cells. Subsequently, RCAN1.4 was identified as a target of miR-619-5p and the suppression of RCAN1.4 inhibited the angiogenesis of HUVEC cells. Furthermore, the suppression of RCAN1.4 by miR-619-5p promoted the cellular mobility and proliferation in in vitro, as well as in vivo, tumor growth and metastasis in non-small cell lung cancer (NSCLC) cells. In patients with non-small cell lung cancer (NSCLC), the level of RCAN1.4 expression was significantly lower in tumor tissues than normal lung tissues, whereas the level of miR-619-5p was a significant higher in tumor tissue than normal lung tissues. We also found that there was a significant inverse correlation between the expression of miR-619-5p and that of RCAN1.4. In addition, miR-619-5p expression was higher in exosomes isolated from plasma of patients with NSCLC than normal. Finally, hypoxia condition led to enhance the amount of miR-619-5p within NSCLC cells-derived exosomes. In conclusion, exosomal miR-619-5p promotes tumor growth and metastasis of NSCLC by regulating RCAN1.4 in endothelial and tumor cells, and it can serve as a diagnosis and prognostic indicator for NSCLC patients.

Citation Format: Sang Yong Jung, Dong Ha Kim, Yong-Hee Cho, Yun Jung Choi, Seon Ye Kim, Ki Jung Sung, Chae Won Lee, Hyeon Jeong Lee, Chang-Min Choi, Jae Cheol Lee, Jin Kyung Rho. Tumor-derived exosomal miR-619-5p promotes tumor angiogenesis and metastasis through the inhibition of RCAN1.4 [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 2528.