bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4355: Altered acetate metabolism in cisplatin resistant bladder cancer
Authors: Kim Jayoung
Published: August 2019

Introduction and Objective: Cisplatin is an important chemotherapeutic agent against metastatic bladder cancer, but resistance often limits its usage. With the recent recognition of lipid metabolic alterations in bladder cancers, we studied the metabolic implications of cisplatin resistance using cisplatin-sensitive (T24S) and resistant (T24R) bladder cancer cells.

Methods: To test whether there should be differences in metabolism and metabolism-associated pathways between cisplatin-sensitive and resistant bladder cancer cells, we applied the live metabolomics approach that we recently developed to isogenic bladder cancer cell lines T24S (cisplatin sensitive) and T24R (cisplatin resistant). The metabolites generated from 13C-glucose tracer were monitored with 2D 1H-13C HSQC NMR in real time.

Results: Real-time live metabolomics revealed that T24R cells consume more glucose, leading to higher production of glucose-derived acetate and fatty acids. Along with the activation of general metabolic regulators, enzymes involved in acetate usage (ACSS2) and fatty acid synthesis (ACC) and a precursor for fatty acid synthesis (acetyl-CoA) were elevated in T24R cells. Consistently, metabolic analysis with 13C isotope revealed that T24R cells preferred glucose to acetate as the exogenous carbon source for the increased fatty acid synthesis, contrary to T24S cells. In addition, ACSS2, rather than the well-established ACLY, was the key enzyme that supplies acetyl-CoA in T24R cells through glucose-derived endogenous acetate. The relevance of ACSS2 in cisplatin resistance was further confirmed by the abrogation of resistance by an ACSS2 inhibitor and, finally, by the higher expression of ACSS2 in the patient tissues with cisplatin resistance.

Conclusions: Our results may help improve the treatment options for chemoresistant bladder cancer patients and provide possible vulnerability targets to overcome the resistance.

Citation Format: Jayoung Kim. Altered acetate metabolism in cisplatin resistant bladder 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 4355.
Abstract 4362: Glucose transporter 1 regulates cell glycolysis and proliferation in gastrointestinal stromal tumor and its clinicopathological significance
Authors: Iwatsuki Masaaki , Sawayama Hiroshi , Kuroda Daisuke , Koga Yuki , Yamashita Kohei , Harada Kazuto , Iwagami Shiro , Eto Kojiro , Ishimoto Takatsugu , Baba Yoshifumi , Yoshida Naoya , Ajani Jaffer A. , Baba Hideo
Published: August 2019

Background: The unique features that cancer cells prefer glycolysis rather than mitochondrial respiration despite of the presence of oxygen have been a potential target for cancer treatment. GIST is well-known as to demonstrate intense metabolic activity and avid glucose uptake on 18F-FDG PET scanning. In this study, we focused on glucose transporter 1 (GLUT 1), a key glycolytic enzyme which plays critical roles in several cancer type. The aim of this study was to examine the clinicopathological significance of GLUT1 expression in GIST and to characterize the molecular mechanism of cell glycolysis and proliferation by GIST cell.

Methods: We determined the expression level of GLUT1 in 65 GIST cases, correlated those values with clinicopathological features. Furthermore, we investigated that the alteration of GLUT1 can influence on glycolysis, cell proliferation, and cell cycle distribution by extracellular flux analyzer, CCK-8 assay and flow cytometry, respectively in GIST cell in vitro.

Results: Fifty-one (78.5%) cases were placed in the high GLUT1 expression group. In the high GLUT1 expression group, tumor size was significantly larger compared to the low GLUT1 expression group (P=0.02). There are significantly more high-risk cases by modified-Fletcher classification in GLUT1 expression group (P< 0.01). The patients with high GLUT1 expression significantly exhibited a higher SUVmax value (P< 0.01). Compared with the low GLUT1 expression group, the high expression group showed a significantly poorer prognosis in terms of recurrence-free survival. In vitro, GLUT1-specific siRNA suppressed cell proliferation and led to increase in G1 phase arrest. Also, knockdown of GLUT1 cells reduced glycolysis as a function of ECAR by extracellular flux analyzer.

Conclusion: We showed that GLUT1 plays a critical role in GIST by regulating glycolysis and proliferation. Our study suggests that glucose transporter can be a novel target of anticancer therapeutics mediating glucose metabolism in GIST.

Citation Format: Masaaki Iwatsuki, Hiroshi Sawayama, Daisuke Kuroda, Yuki Koga, Kohei Yamashita, Kazuto Harada, Shiro Iwagami, Kojiro Eto, Takatsugu Ishimoto, Yoshifumi Baba, Naoya Yoshida, Jaffer A. Ajani, Hideo Baba. Glucose transporter 1 regulates cell glycolysis and proliferation in gastrointestinal stromal tumor and its clinicopathological significance [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 4362.
Abstract 4371: To investigate the role of heme flux and function in growth and progression of NSCLC in vivo
Authors: Ashrafi Adnin , Ghosh Poorva , Sohoni Sagar , Zhang Li
Published: August 2019

Lung cancer is the leading cause of cancer-related mortality and about 85% of the cases are non-small cell lung cancer (NSCLC). Previous research from our lab has shown that the levels of rate limiting heme biosynthetic enzyme ALAS1, heme uptake proteins HCP1 and HRG1 and oxygen utilizing hemoproteins are strongly elevated in NSCLC cells. Furthermore, specific inhibition of heme synthesis using a well-known inhibitor suppresses NSCLC cell proliferation, colony formation and migration compared to normal cells. These results suggest that heme flux and function is important for NSCLC growth and progression.

Lentiviral particles overexpressing eGFP, Alas1, HRG1, and HCP1 were used to transfect NSCLC cell lines. Tumorigenic properties of these cells were analyzed in vitro using invasion, migration and colony formation assay. These cell lines were used to implant subcutaneous xenografts in NOD/SCID mice (n=6). Tumor growth was monitored using Bioluminescence imaging (BLI) and Vernier caliper measurements over a period of five weeks. Mice were sacrificed, tumor tissues were harvested and processed and embedded into FFPE blocks. The tissue sections were then used for histopathological analysis (Hematoxylin and Eosin staining) and immunohistochemistry experiments.

Our data show a significant increase in invasion, migration and colony formation properties of cells lines overexpressing Alas1, HRG1 and HCP1 compared to eGFP overexpressing cell lines (Control) in vitro. At week 5, BLI radiance (total photons/seconds), tumor volumes and masses of Alas1, HCP1 and HRG1 overexpression subcutaneous tumor xenografts were significantly higher compared to eGFP overexpression controls. This was further corroborated by H and E staining.

Our results demonstrate that overexpression of heme synthesis and uptake proteins causes an increase in tumorigenic properties of NSCLC cells in vivo. Thus, heme flux and function plays an important role in growth and progression of NSCLC. Further studies are underway to discern the molecular mechanisms underlying the role of heme in lung tumor growth.

Citation Format: Adnin Ashrafi, Poorva Ghosh, Sagar Sohoni, Li Zhang. To investigate the role of heme flux and function in growth and progression of NSCLC 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 4371.
Abstract 4378: The association of enzymes related to cholesterol esters metabolism with hepatocellular carcinoma
Authors: Liu Zhirong , Shalaby Akram , Subramony Charu , Mao Jinghe , Zhou Xinchun
Published: August 2019

Objectives: The roles of cholesteryl esters (CE) in the pathogenesis of hepatocellular carcinoma (HCC) have not been completely elucidated. This study is aimed to investigate differences in the expression level of enzymes involving in synthesis and degradation of CEs between benign hapatic tissues (BHT) and HCC, and between male and female patients from African American (AA) and Caucasian American (CA).

Methods: TMAs were constructed with 97 BHT and 91 HCC samples from patients enrolled in the University of Mississippi Medical Center from 2010 to 2017. Immunochemistry (IHC) was performed on the TMAs for enzymes involved in CE synthesis (acyl CoA:cholesterol acyltransferase 1, ACAT1 and acyl CoA:cholesterol acyltransferase 2, ACAT2), and in CE degradation (lysosomal acid lipase, LAL and neutral cholesterol ester hydrolase 1, NCEH1) following the manufacturer’s instructions. The expression level for each enzyme was expressed as IHC score calculated with a scoring system. Two-tailed Student’s t-test was used to compare the difference in the expression level of each enzyme between two groups. Significant p value was set at p<0.05.

Results: The IHC score of ACAT2 was significantly higher in HCC than in BHT in all population (6.21±2.42 vs (4.89.4±2.40, P=0.0006) and in stratified male population (6.15±2.33 vs 5.08±2.32, p=0.046), but not in stratified women, AA and CA populations. There was no significant difference in IHC score for ACAT1 between HCC and BHT in all population and populations stratified by race and gender. The IHC score of LAL was significantly higher in HCC as compared to BHT in all population (3.57±2.31 vs 2.40±1.30, P =0.0001) and in male population (3.33±2.01 vs 2.39±1.16, p=0.015), but not in stratified women, AA or CA population. The IHC score of NCEH1 was not significantly different between HCC and BHT in all studied population and population stratified by gender. However, it was disparate between AA and CA populations: The IHC score of NCEH1 was significantly lower in HCC than in BHT in AA population (5.59±2.04 vs 6.66±1.74, p=0.031) but it was higher in HCC than in BHT in CA population (6.84±1.96 vs 7.12±1.70, p>0.05).

Conclusions: These results suggest that alterations in the expression levels of enzymes involving CE metabolism are associated with pathogenesis, progression and disparities in race and gender of HCC. This study also suggest that ACAT2 predominates CE synthesis as compared to ACAT1, and that LAL plays major role in CE degradation as compared to NCEH1 in association with HCC.

Citation Format: Zhirong Liu, Akram Shalaby, Charu Subramony, Jinghe Mao, Xinchun Zhou. The association of enzymes related to cholesterol esters metabolism with hepatocellular carcinoma [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 4378.
Abstract 4380: LSR promotes cell proliferation and migration subsequent to uptake of fatty acid in epithelial ovarian cancer
Authors: Hiramatsu Kosuke , Serada Satoshi , Fujimoto Minoru , Ueda Yutaka , Kimura Tadashi , Naka Tetsuji
Published: August 2019

Background Previously, we identified lipolysis-stimulated lipoprotein receptor (LSR) as a new therapeutic target of epithelial ovarian cancer (EOC), and we reported anti-cancer effect of our newly developed monoclonal antibody (mAb) against LSR-positive EOC in vitro and in vivo. We also demonstrated that LSR took in triglyceride-rich protein and contributed cell proliferation, however, lipid metabolic pathway via LSR is still unclear. Thus, we aimed to reveal the function of LSR on lipid metabolism in EOC cells.

Methods We investigated the activation of beta-oxidation, ATP production, cell proliferation and cell migration via LSR after administration of fatty acid (FA) using LSR-positive EOC cells and LSR-knockout EOC cells. Moreover, we also investigated anti-cancer effect of our anti-LSR mAb against lipid metabolism via LSR.

Results In LSR-positive EOC cells, FA administration increased intracellular lipid accumulation. In glucose restricted environment, LSR promoted FA uptake and promoted ATP production (p < 0.05). In addition, FA promoted cell proliferation and cell migration in LSR-positive EOC cells (p < 0.05). These results suggest that LSR promoted FA uptake in glucose restricted environment and also promoted lipid metabolism including beta-oxidation, TCA cycle and electron transport system leading to ATP production. Finally, our anti-LSR mAb inhibited these lipid metabolic pathways (p < 0.05).

Conclusions Fatty acid uptake via LSR in glucose restricted environment contributed to cell proliferation and migration subsequent to activation of lipid metabolism, and anti-LSR mAb inhibited these processes. LSR might contribute to cancer spread and metastasis of EOC.

Citation Format: Kosuke Hiramatsu, Satoshi Serada, Minoru Fujimoto, Yutaka Ueda, Tadashi Kimura, Tetsuji Naka. LSR promotes cell proliferation and migration subsequent to uptake of fatty acid in epithelial ovarian 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 4380.
Abstract 4393: Integrative proteomic analysis of prostate cancer reveals distinct regulation of RNA binding proteins during disease progression
Authors: Scaravilli Mauro , Kohvakka Annika , Ruusuvuori Pekka , Afyounian Ebrahim , Nykter Matti , Visakorpi Tapio , Latonen Leena
Published: August 2019

To understand the etiology of the disease, and to find novel and more specific drug targets, the driver mutations and expressional changes in prostate cancer have been examined through extensive genomic and transcriptomic characterization. Although significant insight has been gained through these efforts, it is clear that not all molecular alterations influencing the tumor outcome can be captured through these approaches, and that a comprehensive understanding of the molecular events in cancer require thorough investigation of the proteome.

To understand the functional consequences of genetic and transcriptional aberrations in prostate cancer, we aimed to reveal the proteomic changes during disease formation and progression. We performed high throughput mass spectrometry on clinical tissue samples of benign prostatic hyperplasia (BPH), untreated primary prostate cancer (PC) and castration resistant prostate cancer (CRPC). We performed an integrative analysis of the proteomic data with gene copy number, DNA methylation, and RNA expression data from the same samples. Furthermore, proteomic events correlating with the androgen receptor (AR) status of the tumors were analysed.

We uncovered previously unrecognized molecular and pathway events and several novel AR-associated events in the prostate cancer proteomes to study further. We found significant changes in expression of RNA-binding proteins during disease formation and progression. Examining the relationship of RNA binding proteins at the RNA and protein expression level reveal that while many RNA binding proteins exhibit correlation between the expression levels, some seem regulated at the posttranslational level. Two RNA binding proteins, TDP-43 and FUS, which regulated at the protein, but not at RNA level during prostate cancer progression, show opposite behavior during disease progression and correlation with AR status of the tumors. In cultured prostate cancer cell models, we show that these proteins have specific, but divergent interactions with AR at the RNA and protein levels, and that they contribute differentially to AR activity-mediated responses. Thus, these proteins may significantly contribute to prostate cancer molecular evolution and may pinpoint possible targetable pathways in future prostate cancer therapy.

Citation Format: Mauro Scaravilli, Annika Kohvakka, Pekka Ruusuvuori, Ebrahim Afyounian, Matti Nykter, Tapio Visakorpi, Leena Latonen. Integrative proteomic analysis of prostate cancer reveals distinct regulation of RNA binding proteins during disease progression [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 4393.
Abstract 4403: Regulation of SATB2 via miR-31 in Ni-induced malignant cell transformation
Authors: Chen Qiao Yi , Zhu Yusha , Jordan Ashley , Li Jinquan , Sun Hong , Kluz Thomas , Costa Max
Published: August 2019

The special AT-rich DNA binding protein (SATB2) is a nuclear matrix associated protein and an important transcription factor for biological development, gene regulation, and chromatin remodeling. Under normal circumstances, SATB2 is not expressed in most cells, including epithelial cells of the lung. However, aberrant regulation of SATB2 has been found in various types of cancers including lung, colon, prostate, breast, gastric, and liver. Emerging studies have linked the possible role of SATB2 in carcinogenic metal-induced cell transformation. In fact, our previous microarray analysis showed that SATB2 is strongly induced in Ni-transformed cells. As an IARC Class I Human Carcinogen, there is well-established link between chronic nickel exposure and lung cancer; however, despite of considerable research efforts in regards to the epigenetic mechanisms, little is known about the role of microRNAs in Ni-induced carcinogenesis. As post-transcriptional regulators, miRNAs have great importance in maintaining normal cellular development. In particular, miR-31 is one of the most abundant types of miRNAs and commonly studied in disease development, including cancers of the lung, breast, and liver. Previous studies have shown the miR-31 can directly target the homeobox gene SATB, and here we report that miR-31 is capable of regulating SATB2 in Ni-induced tumorigenesis. First, our results confirmed that knocking down SATB2 in Ni-transformed BEAS-2B cells via shRNA significantly reduced the rate of migration, anchorage-independent growth. Notably, compared to scramble control shRNA transfected cells, SATB2 knockdown cells did not generate tumor growth in mice xenograft model. These results highlight the oncogenic role of SATB2 in cell transformation. In addition, our data indicate that both acute Ni-treated and Ni-transformed cells demonstrated significant reduction in miR-31 expression. Furthermore, we show that overexpressing miR-31 in Ni-transformed cells reduced rates of migration, anchorage-independent growth, and cellular invasion, indicating the importance of miR-31 in Ni-induced BEAS-2B cell transformation. Together, our results provide a novel mechanistic pathway for Ni-induced carcinogenesis, and add to the pool of existing research on the implications of SATB2 and miR-31 in tumorigenesis.

Citation Format: Qiao Yi Chen, Yusha Zhu, Ashley Jordan, Jinquan Li, Hong Sun, Thomas Kluz, Max Costa. Regulation of SATB2 via miR-31 in Ni-induced malignant cell transformation [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 4403.
Abstract 4412: Piperine enhances the efficacy of 5 FU against liver cancer
Authors: Anwanwan David , Singh Santosh Kumar , Lillard James W. , Mishra Manoj K. , Singh Rajesh
Published: August 2019

Liver cancer is one of the most devastating malignancies. Hepatocellular carcinoma (HCC) accounts for >90% of primary liver malignancies and is the third-leading cause of cancer-related deaths worldwide. Systemic chemotherapy is one of the most important treatment modalities for advanced HCC. Unfortunately, long-term treatment acquired resistance in HCC due to the activation of drug efflux pumps. A natural compound in combination with chemotherapeutic drugs is a potential approach to combat the side effects. In this study, we have demonstrated whether piperine (natural compound) could increase the effect of 5-FU in HCC cells. The cell viability (MTT) assay was performed to determine the optimal IC50 values of Piperine, 5-FU alone and in combination. Immunofluorescence study confirmed the effectiveness of combined drug; dead nuclei were found more within the cells treated by the combination compared to piperine or 5-FU alone. The combined drug treatment was most effective in inhibiting proliferation and inducing apoptosis; accordingly, it showed noticeable increases in pro-apoptotic (BAK, BID), PARP and decreases in anti-apoptotic genes (BCL-XL) gene in a western blot and RT-PCR analysis. Further, in SK-HEP-1 cells, the combination upregulated the expression of cell cycle inhibitors (p21WAF1/CIP1, p27KIP), which, in turn, inhibited the expression of CDK2/ Cyclin D1 complex. Therefore, it blocks the transition of cells into G0/G1 to S phase. Taken together, we observed that the combination of 5-FU and Piperine resulted in a significant cytotoxicity and apoptosis as compared to alone. Our results suggest that piperine can be a potential agent in enhancing the efficacy of 5-FU in liver cancer treatment. These finding highlights the promising role of natural bioactive compounds and provides the rationale for further transitional researches.

Citation Format: David Anwanwan, Santosh Kumar Singh, James W. Lillard, Manoj K. Mishra, Rajesh Singh. Piperine enhances the efficacy of 5 FU against liver 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 4412.
Abstract 4433: Novel and highly selective CDK9 inhibitors suppress proliferation of triple negative breast cancer (TNBC) cells in vitro
Authors: Winter Jean M. , Mustafa Ebtihal H. , Wang Shudong , Selth Luke A. , Hickey Theresa E. , Tilley Wayne D.
Published: August 2019

This study evaluates the efficacy of two newly developed selective CDK9 inhibitors (CDK9i) across a panel of TNBC cell lines. MDA-MB-453, MFM-223, MDA-MB-468 and MDA-MB-231 TNBC cells were treated with increasing concentrations of two novel and highly selective CDK9 inhibitors and the effect on proliferation, apoptosis and expression of CDK9 targets determined. MDA-MB-453 and -468 cells showed significant growth inhibition with as little as 150nM of CDK9i, evident 3 days after commencement of treatment. Both MDA-MB-231 and MFM-223 cells were less sensitive to the CDK9 inhibitors, with MDA-231 cells requiring at least 300nM to suppress growth. MFM-223 cells did not demonstrate any growth inhibition after 7 days of culture with CDK9i concentrations up to 1.2uM. Protein expression of CDK9 targets, including RNA Polymerase II (RNAPII), phosphorylated-RNAPII, the proto-oncogene C-MYC, and apoptotic marker cleaved caspase-3, were examined by Western blot after optimal CDK9i exposure across each cell line. CDK9i suppressed phosphorylated-RNAPII, but not total RNAPII, indicative of targeted CDK9 inhibition. The master transcription factor C-MYC, which is highly expressed in TNBC, was downregulated, and cleaved-caspase-3 was upregulated with CDK9i treatment. These data demonstrate cell specific efficacy of novel CDK9 inhibitors in cell line models of TNBC via transcriptional suppression of proto-oncogenes and upregulation of apoptotic pathways. Future studies will identify molecular markers of response to CDK9 inhibition and evaluate these novel inhibitors in TNBC patient derived xenografts.

Citation Format: Jean M. Winter, Ebtihal H. Mustafa, Shudong Wang, Luke A. Selth, Theresa E. Hickey, Wayne D. Tilley. Novel and highly selective CDK9 inhibitors suppress proliferation of triple negative breast cancer (TNBC) cells in vitro [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 4433.
Abstract 4435: Molecular mechanism of a novel covalent allosteric inhibitor of SUMO E1 activating enzyme
Authors: Lyu Zongyang , Yuan Lingmin , Williams Katelyn M. , Atkison James H. , Olsen Shaun K.
Published: August 2019

Post-translational modification of proteins by ubiquitin and ubiquitin-like proteins (collectively termed Ubls) proceeds through the sequential interactions and activities of parallel cascades of enzymes that are structurally and mechanistically related. E1 enzymes initiate Ub/Ubl conjugation cascades by catalyzing the ATP-dependent activation and transfer of their cognate Ub/Ubl to E2-conjugating enzymes, which then function with an array of E3 ligases to catalyze the formation of an isopeptide bond linking the Ub/Ubl to target proteins. Aberrations in Ub/Ubl modifications are associated with the pathogenesis of a wide range of life-threatening diseases, which has resulted in several Ubl E1 enzymes emerging as attractive targets for the development of small molecule therapeutics. We present the crystal structure of the E1 enzyme for the Ubl SUMO in complex with a small molecule representing a new class of Ubl E1 enzyme inhibitor. The structure reveals that the covalent allosteric inhibitor targets a cryptic pocket distinct from the active site that is completely buried in all previous SUMO E1 structures and that inhibitor binding is accompanied by a network of structural changes that altogether lock the enzyme in a previously unobserved inactive conformation. Altogether our studies provide the molecular mechanisms of inhibition SUMO E1 specificity and establishes a framework for the development of molecules targeting other Ubl E1s at a newly discovered cryptic allosteric site.

Supported by NIH R01GM115568, F30CA216921, T32CA193201, and a Hollings Cancer Center Postdoctoral Fellowship.

Citation Format: Zongyang Lyu, Lingmin Yuan, Katelyn M. Williams, James H. Atkison, Shaun K. Olsen. Molecular mechanism of a novel covalent allosteric inhibitor of SUMO E1 activating enzyme [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 4435.
Abstract 4443: Targeting E3 ligase PJA1 via TGF-β pathway in hepatocellular carcinoma
Authors: Ohshiro Kazufumi , Chen Jian , Jogunoori Wilma , Deng Chu-Xia , Mishra Bibhuti , Li Shulin , Mishra Lopa
Published: August 2019

Background: RING-finger E3 ligases are instrumental in the regulation of inflammatory cascades, apoptosis, and cancer. However, their roles are relatively unknown for specific pathways such as TGF-β/Smad signaling that when disrupted, drive hepatocellular carcinoma (HCC). We, therefore, analyzed the TCGA database for the TGF-β pathway associated 22 E3 ligases, and identified mRNA alterations in 55% of tumors, most prominently for UCHL5 (16.4%), PJA (12.7%), WWP2 (11.8%), SKP2 (9.1%), SMURF1 and SMURF2 (8.2 and 9.1%, respectively), ITCH (6.4%) and KEAP1 (6.4%). We previously uncovered increased PJA1 expression with loss of TGF-β substrates Smad3 and β2SP in TGF-β deficient mice (β2SP+/-/Smad3+/- mice), which spontaneously develop characteristics of a human stem cell syndrome, and HCC by 12-15 months. Here, we report PJA1 oncogenic function in HCC progression. HCC.

Methods & Results: (1) Analyses of primary HCC datasets (91 cases) reveal increased PJA1 correlates with decreased levels of TGF-β/Smad3 and their regulated genes including Smad8 and TGFBR3. (2) To better understand the implications of PJA1-mediated deregulation of Smad3-promoted transcription in HCC, we compared transcriptomes of HepG2 cells silenced by PJA1 shRNA or treated with TGF-β: 1,584 genes including c-FOS were co-up-regulated and 1,279 genes including TERT were co-down-regulated. PJA1 expression in HCC was negatively associated with c-FOS and SERPINE1 expression. (3) PJA1 interacts with the Smad3 MH2 and Linker domains and multiple domains of β2SP to promote ubiquitin-mediated phosphor-Smad3 degradation in a TGF-β dependent manner, resulting in decreased TGF-β target gene activity. (4) We found that PJA1 expression in HCC is negatively associated with c-FOS and SERPINE1 expression. (5) Overexpression of a RING-domain-deleted PJA1 mutant reduced the proliferation rate in HCC cells and PJA1 knockdown significantly decreased anchorage-independent growth and tumorigenicity of HCC cells. (6) Our results also demonstrate that PJA1 promotes liver cancer stem cell formation in Smad3+/- mice.

Conclusions: This study demonstrates that loss of expression of β2SP and Smad3 through PJA1 could play an important role in HCC progression and that PJA1 is a potential novel therapeutic target for this lethal disease.

Citation Format: Kazufumi Ohshiro, Jian Chen, Wilma Jogunoori, Chu-Xia Deng, Bibhuti Mishra, Shulin Li, Lopa Mishra. Targeting E3 ligase PJA1 via TGF-β pathway in hepatocellular carcinoma [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 4443.
Abstract 4450: Novel transducer array layouts to optimize the treatment of multiple brain metastases with tumor treating fields
Authors: Yesharim Ofir , Naveh Ariel , Bomzon Ze'ev
Published: August 2019

Introduction:

Tumor Treating Fields (TTFields) is an antimitotic cancer treatment approved for the treatment of Glioblastoma Multiforme (GBM). TTFields are delivered using 2 pairs of transducer arrays positioned on the shaved scalp of the GBM patient to optimize the TTFields dose delivered to the tumor. This results in an increased dose of TTFields to the affected tumor region, whilst reducing the intensity of the fields to other regions. Brain metastases, or secondary brain tumors, occur in 10 to 30 percent of adults with cancer. Cancer types most likely to cause brain metastases are lung, breast, colon, kidney and melanoma. When using TTFields to treat brain metastases, it may be desirable to deliver TTFields at therapeutic intensities to the entire brain, in order to treat multiple rather than a single lesion. We investigated novel transducer array layouts designed to deliver a uniform distribution of TTFields to the entire brain.

Methods

Computer simulations were used to calculate the field distributions generated by different array layouts. The simulations utilized a realistic computerized head model of a 40+ years old human male prepared in-house from a T1 MRI series. Using Sim4Life v3.0 (ZMT Zürich), we simulated the delivery of TTFields using pairs of array layouts placed at different locations on the head and neck. To analyze the field distributions, the brain was divided into five regions: (1) the cerebellum, brain stem and other infra-tentorial anatomical regions; and (2-5) the four quadrants of the cerebrum. The mean and median field intensities in the five regions generated by each layout were calculated and compared.

Results

Median intensities between 1.5 V/cm to 1.7 V/cm within all regions were achieved using a layout in which one pair of arrays was placed on the right temple and left scapula, while the second pair was placed on the left temple and right scapula. This layout yielded a uniform intensity distribution within the brain.

Conclusion

We have identified a novel TTFields array layout that could potentially be used to treat the entire brain with sufficiently high intensity to treat for brain metastases. The ongoing Phase 3 METIS study [NCT02831959 is investigating radiosurgery with TTFields for 1-10 brain metastases from non-small cell lung cancer.

Citation Format: Ofir Yesharim, Ariel Naveh, Ze'ev Bomzon. Novel transducer array layouts to optimize the treatment of multiple brain metastases with tumor treating fields [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 4450.
Abstract 4452: GNE-0011, a novel monovalent BRD4 degrader
Authors: Blake Robert A.
Published: August 2019

The bromodomain and extra-terminal (BET) family of proteins (BRD2, 3, 4 and T) has been the focus of an emerging paradigm in drug mechanism of action, whereby a drug is designed to trigger the proteolytic degradation of the target protein. The current design of protein degraders is typically based on bivalent molecules consisting of a ligand for the target protein and a ligand for a ubiquitin ligase (such as VHL, CRBN or XIAP). The degrader causes the association of the target protein with the E3 ubiquitin ligase and its subsequent ubiquitination and degradation. Due to their bivalent design such drugs typically have a higher molecular weight than classic small molecule drugs and may present some non-ideal properties as drugs. An alternative strategy for designing drugs that act through the degradation of the target protein is exemplified by the group of drugs termed SERDs (selective estrogen receptor degraders), for example fulvestrant. The molecular structures of SERDs are typically designed around an estrogen receptor ligand and a “degradation tail”, whose presence results in the degradation of the estrogen receptor. We will discuss the development of GNE-0011, which represents a novel class of monovalent BRD4 degrader that uses a “degradation tail” strategy.

Citation Format: Robert A. Blake. GNE-0011, a novel monovalent BRD4 degrader [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 4452.
Abstract 4457: Open science medicinal chemistry: Towards a treatment for DIPG
Authors: Cramp Sue , Hamblin Nicole , O'Meara Jeff , Edwards Aled , Roberts Owen , Bullock Alex , Brennan Paul
Published: August 2019

Meds4Kids Pharma (M4K)1 is pioneering an open science approach to drug discovery, focussed on the discovery and development of small molecule therapeutics for orphan paediatric cancers. M4K seeks to test the hypothesis that an open science framework can be successfully applied not only to accelerate basic science, using the collective knowledge of the scientific community at large, but also to take an innovative new drug candidate all the way from discovery and clinical proof-of-concept through to product registration, by making use of regulatory data protections and market incentives. Since late 2017, Charles River Early Discovery has been providing in kind drug discovery services to help progress these efforts, including medicinal and synthetic chemistry. The first M4K project aims to generate an orally-available, brain-penetrant therapeutic to treat the diffuse intrinsic pontine glioma (DIPG). DIPG is a rare, aggressive and uniformly fatal childhood brain cancer with a median survival time of 9-12 months and for which there are currently no effective drug treatments. The disease has been shown to be associated with mutations in the ACVR1 gene (activin A receptor, type 1) also known as ALK2 kinase. Early support for the therapeutic hypothesis that an inhibitor of ALK2 kinase would have clinical benefit in DIPG, came from in vivo studies with non-selective ALK2 kinase inhibitors, which both killed DIPG cell lines harbouring the ALK2 mutation and extended lifespan in xenograft mouse models.2 Working collaboratively with M4K and their open science partners, we have made excellent progress towards the identification of potent, selective, brain penetrant ALK2 inhibitors starting from the known inhibitor LDN-214117, previously described for FOP (fibrodysplasia ossificans progressiva).3 Our current lead compound has an excellent in vitro and in vivo profile showing high oral bioavailability and brain penetration in a mouse PK study and is progressing to proof of concept studies. In addition we are progressing with the identification of a back-up series. References: 1. https://m4kpharma.com/ 2. Carvalho et. al. Neuro-Oncology, Volume 18, Issue suppl_6, 1 November 2016, Pages vi154, https://doi.org/10.1093/neuonc/now212.639 3. Mohedas et. al., J. Med Chem, 2014, 57 (19), 7900

Citation Format: Sue Cramp, Nicole Hamblin, Jeff O'Meara, Aled Edwards, Owen Roberts, Alex Bullock, Paul Brennan. Open science medicinal chemistry: Towards a treatment for DIPG [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 4457.
Abstract 4463: Tumor-derived cell-free DNA from bronchoalveolar lavage fluid (BALF): A potential liquid biopsy analysis in lung cancer patients
Authors: Gong Yuhua , Zhang Xinyu , Li Chun , Wang Weiran , Ye Maosong , Zhao Yancheng , Yi Xin , Xu Yaping , Hu Qin , Guan Yanfang , Yang Ling , Xia Xuefeng , Zhou Min , Huang Jian'an , Zhang Hua , Ren Tao , Shen Qian , Wang Kai , Hou Yingyong , Zhang Xin
Published: August 2019

Background: Bronchoalveolar lavage (BAL) is the most common technique for sampling the components of the alveolar space. Here, we evaluated the potential use of BAL fluid (BALF) in liquid biopsy in lung cancer.

Methods: This study enrolled 56 lung cancer patients. And 57 BALF (separated supernatant and precipitate) samples and 56 peripheral blood lymphocytes samples were collected. 57 formalin-fixed Paraffin-embedded (FFPE) tissues were also obtain. We utilized a 1021-gene NGS panel in matched tissue DNA, BALF supernatant cfDNA and BALF precipitate DNA to identify somatic mutations with white blood cell DNA as a germline control.

Results: Mutations were identified in 52 (~91.22%) BALF precipitate samples and 55 (~96.49%) BALF supernatant samples, comparing to 53 (92.98%) in tumor samples. EGFR mutations were observed in 22 BALF precipitate samples and 24 supernatant samples. Among these, 23 were also identified in matched tumor samples. One BALF precipitate samples specific and two BALF supernatant samples specific EGFR mutations were confirmed by Cobas assay. In addition, gene fusions including ALK, ROS1 and RET were found in 6 tumors, 4 BALF precipitate samples and 5 supernatant samples, respectively.

In total, there were 195 mutations identified in the tumor samples, of which 147 (~75.4%) mutations were detected in the matched BALF precipitate samples, 168 (~86.1%) mutations were detected in the matched BALF supernatant samples and 145 (~74.4%) mutations were shared in three types of samples. Tumor mutation burden (TMB) were calculated, and the consistency between tissue and BALF precipitate was 90% and the consistency between tissue and BALF supernatant was 94%.

Conclusions: Liquid biopsy using BALF shown high potential to profiling genetic alterations of patients with lung cancer, which might be implemented and standardized into clinical use.

Citation Format: Yuhua Gong, Xinyu Zhang, Chun Li, Weiran Wang, Maosong Ye, Yancheng Zhao, Xin Yi, Yaping Xu, Qin Hu, Yanfang Guan, Ling Yang, Xuefeng Xia, Min Zhou, Jian'an Huang, Hua Zhang, Tao Ren, Qian Shen, Kai Wang, Yingyong Hou, Xin Zhang. Tumor-derived cell-free DNA from bronchoalveolar lavage fluid (BALF): A potential liquid biopsy analysis in lung cancer patients [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 4463.
Abstract 4467: Survival of patients with microsatellite instable (MSI) metastatic colorectal cancer (mCRC) upon systemic non-immunotherapy
Authors: Wensink G. E. , Elferink M. A. , May A. M. , Mol L. , Hamers P. A. , Punt C. J. , Vink G. R. , Roodhart J. M. , Koopman M.
Published: August 2019

Introduction: Data on survival of patients with MSI mCRC treated with systemic non-immunotherapy are scarce, which complicates the interpretation of recent immunotherapy trials. Our study examined survival in MSI mCRC patients in a retrospective observational cohort.

Methods: We performed a study involving 186 mCRC MSI patients (n=54 from the phase 3 CAIRO, CAIRO2 and CAIRO3 trials and n=132 from The Netherlands Cancer Registry 2015 and 2016). Overall survival from diagnosis of mCRC (OS), from start of first-line (OS1) and second-line therapy (OS2) to death of any cause were examined. Progression-free survival from start of first line (PFS1) and second-line therapy (PFS2) to progression or death of any cause were examined.

Results: Of the 186 patients, 57% were females, median age 67 years, 19% had liver-only metastasis, 31% peritoneal metastasis and 23% underwent metastasectomy. In 113 patients with known BRAF status, 51% had BRAF mutated tumors. The proportion of patients receiving first, second and third-line systemic treatment was 68%, 27% and 7%, respectively. For all patients, median OS was 13.8 months (95% confidence interval [C.I.] 11.8 - 16.3). In patients receiving at least one line of treatment median OS was 15.6 months (13.8 - 19.3), OS1 was 13.9 months (11.6-16.6) and PFS1 was 6.4 months (5.5 - 9.2). In the 48 patients receiving second-line therapy, median OS was 16.5 months (13.9 - 21.5), OS2 was 6.1 months (5.4 - 9.7) and PFS2 was 2.2 months (1.9 - 4.2). Patients starting second-line treatment with a performance score of 0-1, thus eligible for CHECKMATE-142, had an OS2 of 6.7 months (5.0 - 12.1).

Discussion: A direct comparison between our cohort and the CHECKMATE-142 cohort is not possible due to key differences in the patient populations: CHECKMATE-142 patients often received more than one therapy line prior to inclusion and displayed fewer BRAF mutant tumors. However, the survival from start of second-line in our cohort of approximately 7 months is inferior to the published CHECKMATE-142 results, where median OS was not reached after a median follow-up of 1 year in the nivolumab and the nivolumab and ipilimumab cohorts.

Conclusion: We present data on systemic non-immunotherapy in the largest cohort of trial and population-based mCRC MSI patients to date. Our results suggest that immunotherapy may offer a survival benefit for mCRC MSI patients.

Citation Format: G. E. Wensink, M. A. Elferink, A. M. May, L. Mol, P. A. Hamers, C. J. Punt, G. R. Vink, J. M. Roodhart, M. Koopman. Survival of patients with microsatellite instable (MSI) metastatic colorectal cancer (mCRC) upon systemic non-immunotherapy [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 4467.
Abstract 4490: PARP inhibitor efficacy depends on CD8+ T-cell recruitment via the STING pathway in BRCA-deficient models of triple-negative breast cancer
Authors: Pantelidou Constantia , Sonzogni Olmo , Taveira Mateus De Oliveira , Mehta Anita K. , Wang Dan , Kothari Aditi , Li Michelle K. , Visal Tanvi H. , Guerriero Jennifer L. , Wulf Gerburg M. , Shapiro Geoffrey I.
Published: August 2019

Combinatorial clinical trials of PARP inhibitors with immunotherapies are ongoing, yet the immunomodulatory effects of PARP inhibition have been incompletely studied. Here, we sought to dissect the mechanisms underlying PARP inhibitor-induced changes in the tumor microenvironment of BRCA1-deficient triple-negative breast cancer (TNBC). We demonstrate that the PARP inhibitor olaparib induces CD8+ T cell infiltration and activation in vivo, and that depletion of CD8+ T cells severely compromises anti-tumor efficacy. Olaparib-induced T cell recruitment is mediated through activation of the STING/TBK1/IRF3 pathway in tumor and dendritic cells and is more pronounced in HR-deficient compared to HR-proficient TNBC cells. CRISPR-knockout of STING in cancer cells prevents type I IFN production and is sufficient to abolish PARP inhibitor-induced T cell infiltration in vivo. These findings elucidate a novel mechanism of action of PARP inhibitors and provide mechanistic rationale for combining PARP inhibition with immunotherapies for the treatment of TNBC.

Citation Format: Constantia Pantelidou, Olmo Sonzogni, Mateus De Oliveira Taveira, Anita K. Mehta, Dan Wang, Aditi Kothari, Michelle K. Li, Tanvi H. Visal, Jennifer L. Guerriero, Gerburg M. Wulf, Geoffrey I. Shapiro. PARP inhibitor efficacy depends on CD8+ T-cell recruitment via the STING pathway in BRCA-deficient models of triple-negative 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 4490.
Abstract 4501: Loss of miR-21 delays myc-driven prostate cancer progression in the hi-myc transgenic mouse model
Authors: Zennami Kenji , Rafiqi Fatima , Liao Ross , Sealover Kim , Simons Brian , Sumitomo Makoto , Shiroki Ryoichi , Lupold Shawn
Published: August 2019

Introduction and Objectives: MicroRNA 21 (miR-21) is overexpressed in virtually all types of cancers including prostate cancer (PCa). We have previously reported that the androgen receptor (AR) induces miR-21 expression, and that elevated miR-21 is sufficient to drive PCa growth and castration resistance. However, the role of miR-21 in PCa initiation and progression has not been fully elucidated. To determine if the absence of miR-21 inhibits or delays prostate cancer incidence or progression, we crossed miR-21 KO mice with Hi-Myc mice. In this study, we identify that loss of miR-21 delays Myc-driven prostate cancer progression in the Hi-Myc transgenic mouse model.

Methods: miR-21 KO mice were backcrossed to FVB mice to generation F10. Resulting miR-21 WT or KO FVB progeny were crossed with Hi-Myc mice to generate Hi-Myc/miR-21 WT or KO mice. Prostates were harvested at 4 weeks (n=10), 3 months (n=10), 5 months (n=58), 6 months (n=25), 8 months (n=20) of age and prostate weight, histology, miR-21 expression levels and protein expression levels were quantified. Proliferation and apoptosis were examined by immunohistochemistry using Ki-67 and cleaved caspase-3.

Results: Cancerous prostates of Hi-Myc mice showed high miR-21 expression at 5m and 8m, when compared to FVB controls, suggesting high miR-21 expression in the Myc-driven prostate cancer. Hi-Myc/miR-21 KO mice demonstrated a marked reduction in adenocarcinoma when compared to Hi-Myc/miR-21 WT mice at 5m. This is apparent in the reduced weight of effected prostate lobes of Hi-Myc/miR-21 KO versus WT mice at 8m. Protein levels of established miR-21 target genes, PDCD4 and PTEN, were up-regulated in the prostates of Hi-Myc/miR-21 KO mice. Also, IHC showed lower Ki-67 and higher cleaved caspase-3 positive cells in miR-21 KO than WT. However, miR-21 KO did not affect prostatic intraepithelial neoplasia (PIN) development.

Conclusions: Our results demonstrate that miR-21 delays Myc-driven prostate cancer progression through reduced proliferation and enhanced apoptosis, and that endogenous miR-21 suppresses prostatic PDCD4 and PTEN protein expression levels. Therefore, targeting of miR-21 or its downstream target genes may be useful for cancer prevention or cancer therapy.

Citation Format: Kenji Zennami, Fatima Rafiqi, Ross Liao, Kim Sealover, Brian Simons, Makoto Sumitomo, Ryoichi Shiroki, Shawn Lupold. Loss of miR-21 delays myc-driven prostate cancer progression in the hi-myc transgenic mouse model [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 4501.