bluuebunny/crossref_metadata_2025 huggingface dataset:
Induction of Myogenic Differentiation Improves Chemosensitivity of Chemoresistant Cells in Soft-Tissue Sarcoma Cell Lines
Authors: Dawson Lucy E. , D’Agostino Luca , Hakim Abraham A. , Lackman Richard D. , Brown Spencer A. , Sensenig Richard B. , Antonello Zeus A. , Kuzin Igor I.
Published: March 2020

Rhabdomyosarcoma (RMS) and rhabdoid tumors (RT) are rare soft-tissue malignancies with the highest incidence in infants, children, and adolescents. Advanced, recurrent, and/or metastatic RMS and RT exhibit poor response to treatment. One of the main mechanisms behind resistance to treatment is believed to be intratumoral heterogeneity. In this study, we investigated the myogenic determination factor 1 (MYOD1) and Noggin (NOG) markers in an embryonal RMS (ERMS) cell line and an RT cell line and the differential response of the MYOD1 and NOG expressing subpopulations to chemotherapy. Importantly, we found that these markers together identify a subpopulation of cells (MYOD1+ NOG+ cells) with primary resistance to Vincristine and Doxorubicin, two commonly used chemotherapies for ERMS and RT. The chemoresistant MYOD1+ NOG+ cells express markers of undifferentiated cells such as myogenin and ID1. Combination of Vincristine with TPA/GSK126, a drug combination shown to induce differentiation of RMS cell lines, is able to partially overcome MYOD1/NOG cells chemoresistance.
The Biomechanical Relationship between Hallux Valgus Deformity and Metatarsal Pain
Authors: Chang Cheng , Wang Qing-Fu , Guo Jun-Chao , Li Duo-Duo , Fan Yu-Bo , Wen Jian-Min
Published: March 2020

Metatarsal pain is a common pathological outcome in patients with a hallux valgus (HV) deformity. However, the relationship between the degree of HV deformity and metatarsal pain has not been systematically examined. The purpose of the present study was to investigate the correlation between metatarsal pain and the degree of HV deformity. Between October 2017 and September 2018, 512 HV patients (944 feet) participated in an evaluation of their HV angle (HVA) using X-ray images. The participants were divided into four groups corresponding to their HVA (<15°, 15° to 20°, 21° to 40°, or >40°). Load rate, impulse, contact duration, and contact area were measured and recorded as dynamic gait parameters using the RsScan system. Data were evaluated using SPSS statistical software. The visual analog scale (VAS) was used to assess metatarsal pain. For the four HV deformity groups, the peak value of impulse and contact duration was concentrated on the second and third metatarsals (Meta2 and Meta3) (P<0.05); contact area was also shown on metatarsals 1, 2, and 5 (P<0.05). Metatarsal pain on Meta2 had the highest VAS score (VAS: 6.57), followed by Meta3 (Mean VAS: 5.72). In the HV > 40° group, the load location on Meta2 was transferred to Meta1. The percent of pain attributed to Meta2 and Meta3 was also increased in this group. These findings illustrated that metatarsal pain was primarily located on Meta2 and Meta3 in the different degrees of HV deformity. This information can provide the location to target for pain relief and help guide further rehabilitation.
Associations between Aquaglyceroporin Gene Polymorphisms and Risk of Stroke among Patients with Hypertension
Authors: Tu Qingyun , Yan Li , Wang Chun , Han Aohan , Qin Yu , Cui Lan , Xiang Quanyong
Published: March 2020

Background. Dysregulations of AQP7 and AQP9 were found to be related to lipid metabolism abnormality, which had been proven to be one of the mechanisms of stroke. However, limited epidemiological studies explore the associations between AQP7 and AQP9 and the risk of stroke among patients with hypertension in China. Aims. We aimed to investigate the associations between genetic variants in AQP7 and AQP9 and the risk of stroke among patients with hypertension, as well as to explore gene‐gene and gene‐environment interactions. Methods. Baseline blood samples were drawn from 211 cases with stroke and 633 matched controls. Genomic DNA was extracted by a commercially available kit. Genotyping of 5 single nucleotide polymorphisms (SNPs) in AQP7 (rs2989924, rs3758269, and rs2542743) and AQP9 (rs57139208, rs16939881) was performed by the polymerase chain reaction assay with TaqMan probes. Results. Participants with the rs2989924 GG genotype were found to be with a 1.74‐fold increased risk of stroke compared to those with the AA+AG genotype, and this association remained significant after adjustment for potential confounders (odds ratio (OR): 1.74, 95% confidence interval (CI): 1.23‐2.46). The SNP rs3758269 CC+TT genotype was found to be with a 33% decreased risk of stroke after multivariate adjustment (OR: 0.67, 95% CI: 0.45‐0.99) compared to the rs3758269 CC genotype. The significantly increased risk of stroke was prominent among males, patients aged 60 or above, and participants who were overweight and with a harbored genetic variant in SNP rs2989924. After adjusting potential confounders, the SNP rs3758269 CT+TT genotype was found to be significantly associated with a decreased risk of stroke compared to the CC genotype among participants younger than 60 years old or overweight. No statistically significant associations were observed between genotypes of rs2542743, rs57139208, or rs16939881 with the risk of stroke. Neither interactions nor linkage disequilibrium had been observed in this study. Conclusions. This study suggests that SNPs rs2989924 and rs3758269 are associated with the risk of stroke among patients with hypertension, while there were no statistically significant associations between rs2542743, rs57139208, and rs16939881 and the risk of stroke being observed.
USP10 Promotes Proliferation of Hepatocellular Carcinoma by Deubiquitinating and Stabilizing YAP/TAZ
Authors: Zhu Hong , Yan Fangjie , Yuan Tao , Qian Meijia , Zhou Tianyi , Dai Xiaoyang , Cao Ji , Ying Meidan , Dong Xiaowu , He Qiaojun , Yang Bo
Published: March 2020

Yes-associated protein (YAP) and its paralog, transcriptional coactivator with PDZ-binding motif (TAZ), play pivotal roles in promoting the progression of hepatocellular carcinoma. However, the regulatory mechanism underpinning aberrant activation of YAP/TAZ in hepatocellular carcinoma remains unclear. In this study, we globally profiled the contribution of deubiquitinating enzymes (DUB) to both transcriptional activity and protein abundance of YAP/TAZ in hepatocellular carcinoma models and identified ubiquitin-specific peptidase 10 (USP10) as a potent YAP/TAZ-activating DUB. Mechanistically, USP10 directly interacted with and stabilized YAP/TAZ by reverting their proteolytic ubiquitination. Depletion of USP10 enhanced polyubiquitination of YAP/TAZ, promoted their proteasomal degradation, and ultimately arrested the proliferation of hepatocellular carcinoma in vitro and in vivo. Expression levels of USP10 positively correlated with the abundance of YAP/TAZ in hepatocellular carcinoma patient samples as well as in N-nitrosodiethylamine (DEN)-induced liver cancer mice models. Collectively, this study establishes the causal link between USP10 and hyperactivated YAP/TAZ in hepatocellular carcinoma cells and provides a rationale for potential therapeutic interventions in the treatment of patients with hepatocellular carcinoma harboring a high level of YAP/TAZ.

Significance:

These findings identify USP10 as a DUB of YAP/TAZ and its role in hepatocellular carcinoma progression, which may serve as a potential therapeutic target for hepatocellular carcinoma treatment.
FDA Approval Summary: Ado-Trastuzumab Emtansine for the Adjuvant Treatment of HER2-positive Early Breast Cancer
Authors: Wedam Suparna , Fashoyin-Aje Lola , Gao Xin , Bloomquist Erik , Tang Shenghui , Sridhara Rajeshwari , Goldberg Kirsten B. , King-Kallimanis Bellinda L. , Theoret Marc R. , Ibrahim Amna , Amiri-Kordestani Laleh , Pazdur Richard , Beaver Julia A.
Published: March 2020

On May 3, 2019, the FDA granted regular approval to ado-trastuzumab emtansine (KADCYLA), for the adjuvant treatment of patients with HER2-positive early-breast cancer (EBC) who have residual invasive disease after neoadjuvant taxane-based chemotherapy and trastuzumab-based treatment. Approval was based on data from the KATHERINE trial, which randomized patients to receive ado-trastuzumab emtansine or trastuzumab. At 3 years, the event-free rate for invasive disease-free survival in the ado-trastuzumab emtansine arm was 88.3% [95% confidence interval (CI), 85.8–90.7] compared with 77.0% (95% CI, 73.8–80.7) in the trastuzumab arm, (HR, 0.50; 95% CI, 0.39–0.64; P < 0.0001). Results from secondary endpoints, subgroup analyses, and sensitivity analyses generally supported the primary efficacy endpoint results. Common adverse reactions (>25% and higher incidence in ado-trastuzumab emtansine arm) with ado-trastuzumab emtansine were fatigue, nausea, increased transaminases, musculoskeletal pain, hemorrhage, thrombocytopenia, headache, peripheral neuropathy, and arthralgia. Ado-trastuzumab emtansine is the first drug approved for the treatment of patients with residual disease after neoadjuvant treatment and surgery. This article summarizes the FDA review and the data supporting the approval of ado-trastuzumab emtansine as a component of treatment for patients with HER2-positive EBC with residual disease.
Resistance to First-line Osimertinib in EGFR-mutant NSCLC: Tissue is the Issue
Authors: Piotrowska Zofia , Hata Aaron N.
Published: March 2020

Matched pre-/posttreatment tissue biopsies from patients with EGFR-mutant non–small cell lung cancer demonstrate that histologic transformations, including both small-cell carcinoma and squamous transformation, are unexpectedly common among patients progressing on first-line osimertinib. The study highlights the key role of tissue testing and underscores the need for innovative therapeutic approaches to prevent, rather than treat, resistance.

See related article by Schoenfeld et al., p. 2654
A Highly Potent TACC3 Inhibitor as a Novel Anticancer Drug Candidate
Authors: Akbulut Ozge , Lengerli Deniz , Saatci Ozge , Duman Elif , Seker Urartu O.S. , Isik Aynur , Akyol Aytekin , Caliskan Burcu , Banoglu Erden , Sahin Ozgur
Published: March 2020

TACC3, a transforming acidic coiled-coil (TACC) family member, is frequently upregulated in a broad spectrum of cancers, including breast cancer. It plays critical roles in protecting microtubule stability and centrosome integrity that is often dysregulated in cancers; therefore, making TACC3 a highly attractive therapeutic target. Here, we identified a new TACC3-targeting chemotype, BO-264, through the screening of in-house compound collection. Direct interaction between BO-264 and TACC3 was validated by using several biochemical methods, including drug affinity responsive target stability, cellular thermal shift assay, and isothermal titration calorimetry. BO-264 demonstrated superior antiproliferative activity to the two currently reported TACC3 inhibitors, especially in aggressive breast cancer subtypes, basal and HER2+, via spindle assembly checkpoint–dependent mitotic arrest, DNA damage, and apoptosis, while the cytotoxicity against normal breast cells was negligible. Furthermore, BO-264 significantly decreased centrosomal TACC3 during both mitosis and interphase. BO-264 displayed potent antiproliferative activity (∼90% have less than 1 μmol/L GI50 value) in the NCI-60 cell line panel compromising of nine different cancer types. Noteworthy, BO-264 significantly inhibited the growth of cells harboring FGFR3–TACC3 fusion, an oncogenic driver in diverse malignancies. Importantly, its oral administration significantly impaired tumor growth in immunocompromised and immunocompetent breast and colon cancer mouse models, and increased survival without any major toxicity. Finally, TACC3 expression has been identified as strong independent prognostic factor in breast cancer and strongly prognostic in several different cancers. Overall, we identified a novel and highly potent TACC3 inhibitor as a novel potential anticancer agent, inducing spindle abnormalities and mitotic cell death.
Abstract A017: Development and characterization of HER2+ T-DM1-resistant breast PDX models in athymic nude mice
Authors: Moriarty Alyssa , Gamez Lizette , Mashimo Tomoyuki , Papadopoulos Kyriakos , Patnaik Amita , Rasco Drew , Lang Amy , Drengler Ronald , Wick Michael J
Published: March 2020

Background: Ado-trastuzumab emtansine (T-DM1) is an antibody-drug conjugate (ADC) approved for treatment of HER2 positive (HER2+), trastuzumab-resistant breast cancers. While this agent is initially effective, resistance often develops. To aid in developing new therapies for HER2+, T-DM1-resistant breast cancer and better understanding resistance mechanisms, we established PDX models designated ST340, ST1339, and ST2167 from three patients with trastuzumab-resistant HER2+ breast cancer and sensitive to T-DM1 treatment in vivo. A cohort of each model was conditioned in vivo with chronic T-DM1 treatment to generate drug resistance. These models, designated ST340/TDR, ST1339/TDR, and ST2167/TDR were subjected to characterization and efficacy studies, and the results compared with parent models. Methods: ST340, ST1339, and ST2167 were established from patients who responded then progressed on trastuzumab and other therapies. The models were passaged, and cohorts were challenged with chronic T-DM1 treatment to produce drug resistance. The parent and resistant models were subjected to various comparative analyses including receptor expression, RNAseq, and response to various relevant therapies. Results: T-DM1-resistant models retained high HER2 expression and demonstrated receptor staining and histopathology comparable to respective parent lines. Comparison of parent and resistant clones using RNAseq identified several alterations in variants and expression. In vivo T-DM1 reported significant activity towards ST340: %T/C=3%, ST1339: %T/C=-40% and ST2167: %T/C=13% versus control, including tumor regressions in ST1339. T-DM1-resistant models reported the following values following treatment: ST340/TDR: %T/C=65%, ST1339/TDR: %T/C=58% and ST2167/TDR: T/C=91%. Conclusion: We have established and characterized three paired T-DM1-sensitive and resistant breast PDX models, which can be utilized as valuable tools in developing new therapies for T-DM1-resistant patients.

Citation Format: Alyssa Moriarty, Lizette Gamez, Tomoyuki Mashimo, Kyriakos Papadopoulos, Amita Patnaik, Drew Rasco, Amy Lang, Ronald Drengler, Michael J Wick. Development and characterization of HER2+ T-DM1-resistant breast PDX models in athymic nude mice [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A017. doi:10.1158/1535-7163.TARG-19-A017
Abstract A030: Evaluation of phosphoprotein- and transcript-based pharmacodynamic biomarkers in pre-clinical studies of the novel SOS1::KRAS inhibitor BI-3406
Authors: Gerlach Daniel , Savarese Fabio , Hofmann Marco H , Trapani Francesca , Gmachl Michael , Botesteanu Dana-Adriana , Ramharter Jürgen , Arnhof Heribert , Schweifer Norbert , McConnell Darryl B , Kraut Norbert
Published: March 2020

KRAS is among the most frequently mutated oncogenes across all human cancers. Targeting KRAS-mutant cancers either directly or at the effector level has so far failed in the clinic, with adoptive responses and toxicities being the major limiting factors. Nevertheless, direct targeting of the mutant KRAS-G12C allele has shown promising results in early clinical trials. BI-3406 is an inhibitor of the interaction between KRAS and its Guanine Nucleotide Exchange Factor (GEF) SOS1. The efficient loading of both mutant as well as wild-type KRAS with GTP depends on SOS1. We present the activity of the potent and selective SOS1::KRAS inhibitor BI-3406 in monotherapy and combinations in a different presentation at this meeting (Marco H Hofmann et al.). In this work, we focus on the characterization of suitable pharmacodynamic biomarkers in pre-clinical studies. Analysis of the well-studied RAS-MAPK effector p-ERK demonstrates BI-3406 treatment induced modulation in the majority of tested cell lines, independent of their KRAS mutation status. Interestingly, the effects on p-AKT levels, linked to PI3K signaling, were much less pronounced, indicating that the MAPK pathway is the major effector pathway of mutant KRAS singnaling. Confirmatory results were obtained from a technically independent and orthogonal platform. These findings are supported by a genome-wide transcriptome analysis, which revealed well established feedback regulators and p-ERK targets such as DUSP6, EGR1, or ETV5 as robust pharmacodynamic biomarkers in response to SOS1::KRAS inhibitor treatment. Importantly, we also compare biomarker responses of SOS1::KRAS inhibition to direct KRAS-G12C inhibition. In summary, these transcript-based biomarkers add value over conventional downstream signaling protein biomarkers such as p-ERK and will be implemented in future clinical trials.

Citation Format: Daniel Gerlach, Fabio Savarese, Marco H Hofmann, Francesca Trapani, Michael Gmachl, Dana-Adriana Botesteanu, Jürgen Ramharter, Heribert Arnhof, Norbert Schweifer, Darryl B McConnell, Norbert Kraut. Evaluation of phosphoprotein- and transcript-based pharmacodynamic biomarkers in pre-clinical studies of the novel SOS1::KRAS inhibitor BI-3406 [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A030. doi:10.1158/1535-7163.TARG-19-A030
Abstract A058: Dielectric characterization of glioblastoma cancer stem cells
Authors: Bortolozzi Roberta , Mariotto Elena , Rampazzo Elena , Porcù Elena , Saada Sofiane , Provent Thomas , Lalloué Fabrice , Pothier Arnaud , Viola Giampietro , Persano Luca
Published: March 2020

Glioblastoma (GBM) is the most aggressive type of primary brain tumor, characterized by the intrinsic resistance to chemotherapy due to the presence of a highly aggressive Cancer Stem Cell (CSC) sub-population. The coexistence in the same mass of bulk and CSCs stuck at different stages of differentiation and genetic backgrounds prevents counteracting all cells with single treatments. Through the application of a Dielectrophoretic force (DEP) we provide a successful CSC isolation from a mixed cell population, exploiting their physical properties. Using FACS, the expression of specific stem cell and differentiation markers (CD133, Nestin, Sox1, Sox2, GFAP, Doublecortin (DCX) and Ki67) were analyzed in 17 primary cell lines of GBM. CD133 expression was also exploited as a reliable marker for the identification of the CSC subpopulation and thus used for sorting CD133+ and CD133- GBM primary cells by flow cytometry techniques. CD133+ and CD133- cells were characterized for their ability to self-renew and form neurospheres. In all experiments, CD133+ cells have shown a significant increased efficiency of neurosphere generation that was accompanied by a higher self-renewal potential as measured by the limiting dilution assay (LDA) in vitro. The characterization and isolation of patient-derived GBM cells by FACS allowed constituting a cell library with three distinct cell subpopulation types: CD133+/CD133-/Total population, that have been characterized for their specific Electromagnetic (EM) signature. Determination of EM signature was performed using a sensor that applies electromagnetic fields in the MHz regime to selectively electromanipulate cells with dielectrophoresis (DEP) and allows the monitoring of the induced cell motion generated by the field. By scanning the frequency range of interest, the cell DEP signature and the cross over frequency (frequency for which the DEP force becomes null) were identified. We found that CD133+ and CD133- cell display a significantly different crossover frequency. In particular, CD133+ cells are characterized by a lower cross over frequency than both CD133- and total population. This data suggest a non obvious correlation between crossover frequency and CSCs phenotype which would be potentially exploited to set up a label free system able to efficiently identify and physically separate subpopulation of cells enriched for CSC characteristics.

Citation Format: Roberta Bortolozzi, Elena Mariotto, Elena Rampazzo, Elena Porcù, Sofiane Saada, Thomas Provent, Fabrice Lalloué, Arnaud Pothier, Giampietro Viola, Luca Persano. Dielectric characterization of glioblastoma cancer stem cells [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A058. doi:10.1158/1535-7163.TARG-19-A058
Abstract A064: MK2 inhibition stabilizes wild-type and mutated p53 in glioblastoma cells and leads to different cellular responses
Authors: Phoa Athena Frances , Recasens Ariadna , Munoz Lenka
Published: March 2020

MAPK-activated protein kinase 2 (MK2) has diverse roles in inflammation and cancer. In this study we show that in glioblastoma cells, MK2 plays a role in the degradation of p53, the main determinant of cell fate. Importantly, we provide evidence that the MK2-dependent regulation of p53 is a common regulatory mechanism for both wild-type and mutated p53. Through the use of both pharmacological inhibitors and MK2 knockout models in glioblastoma cells with different p53 background, we show that MK2 inhibition leads to different cellular responses in response to DNA damage stimuli. In p53 wild-type cells, MK2 inhibition strongly attenuated cell proliferation. This effect was achieved through the deactivation of HDM2 ligase and this led to increased p53 wild-type expression and the transcription of senescent and apoptotic genes. MK2 inhibition increased the stability of p53 mutants, and this is key for their ability to phenotypically exhibit gain-of function properties. Thus, as a destabilizer of wild-type p53, MK2 represents a potential drug target in glioblastomas that contain p53 wild-type. However, the administration of an MK2 inhibitor in cells expressing p53 mutants may cause accelerated disease progression. These findings are highly relevant since p53 mutations occur over 50% of all cancers.

Citation Format: Athena Frances Phoa, Ariadna Recasens, Lenka Munoz. MK2 inhibition stabilizes wild-type and mutated p53 in glioblastoma cells and leads to different cellular responses [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A064. doi:10.1158/1535-7163.TARG-19-A064
Abstract A091: Targeting homologous recombination repair defects in lung cancer
Authors: Muvaffak Asli , Zhou Yinghui , Feng Bin , Wang Sarah , Wang Jing Ju , Ramaswamy Sridhar , Coleman Kevin G
Published: March 2020

Non-Small Cell Lung Cancer (NSCLC) is the leading cause of cancer related mortality worldwide, and accounts for 80-85% of all lung cancer diagnoses. While targeted therapies against epidermal growth factor receptor (EGFR) mutations and anaplastic lymphoma kinase (ALK) fusions have proven to be effective for the treatment of NSCLC, only a fraction of NSCLC patients (i.e. <20%) benefit from these targeted agents. Although immunotherapy approaches, specifically PD-1:PD-L1 blockade, appear to be more broadly efficacious, there are still a substantial proportion of patients who may not benefit from these treatments. To investigate the potential of targeting the DNA Damage Response (DDR) pathway in lung cancer, as an alternative therapeutic approach for these patients we sought to identify whether functionally relevant HRR (Homologous Recombination Repair)-defects could be synthetically lethal with niraparib monotherapy in NSCLC PDX models. Approximately 3% of lung cancers harbor mutations in ATM (i.e. 3.5% in lung adenocarcinomas & 1.4% in lung squamous cell carcinoma), which is involved in HRR as well as multiple other DNA repair and checkpoint functions, but the functional consequences of this alteration remain to be established. Studies in the past few years have demonstrated the potential of targeting the DDR pathway as a promising therapeutic strategy for lung cancer, i.e. in small cell lung cancer (SCLC) where overall response rates to immune checkpoint blockade remain poor. In addition, inhibitors targeting DDR proteins have shown promise in preclinical models of lung cancer, and are under clinical investigation as single agents and in combination with other targeted and immunotherapy agents. In this study, we have evaluated niraparib sensitivity in 57 NSCLC PDX models containing both BRCA and non-BRCA HRR mutations (n=17) as well as HRR WT models (n=40). This analysis demonstrated that niraparib sensitive models include both HRR mutant and HRR WT lung tumors (TGI > 70%). Amongst the PDX models containing a biallelic HRR mutation (ATM, BAP1, RAD51D & XRCC2), only ATM biallelic mutant models were sensitive to niraparib (2 out of 6). Furthermore, none of the of nine monoallelic mutant models (ATM, ATR, BRCA1/2, RAD50 & RAD52) were sensitive to niraparib. Surprisingly, 7.5% (3 out of 40) of the HRR WT PDX models were sensitive to niraparib. Further analysis is required to elucidate the molecular mechanisms driving niraparib sensitivity in HRR WT NSCLC.

Citation Format: Asli Muvaffak, Yinghui Zhou, Bin Feng, Sarah Wang, Jing Ju Wang, Sridhar Ramaswamy, Kevin G Coleman. Targeting homologous recombination repair defects in lung cancer [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A091. doi:10.1158/1535-7163.TARG-19-A091
Abstract A092: Multiplexed measurement of DNA repair capacity for tumor heterogeneity
Authors: Hardie Lakotah , Richer Amanda , Hesselberth Jay
Published: March 2020

Purpose of Study: DNA damage has been shown to have a strong link to the origin and propagation of many cancers. Specific damages to different DNA repair pathways within the cell spell accumulations of DNA damaging events and alterations in normal biochemical activities of the cell. More recently, these biochemical differences have begun to be explored and have demonstrated potential evidence supporting a heterogeneous picture of tumor and cancer pathology. The goal of this project focused upon developing a cellular assay to measure the DNA repair capacities of cancer cells and to apply it at the level of an individual cell. Methods Used: Bulk Hap1 cellular lysate was prepared and incubated with the assay and repair activities were recovered from DNA substrates. Amplification and ILLUMINA sequencing was then used to detail repair capacities. Bioinformatic pipelines systematically compared differences in repair profiles across tested conditions in the optimization of the assay with ATP regeneration and chemotherapy pre-treated cells. Future direction will include the combination of a microfluidic platform for single cell analysis. Summary of Results: The use of a creatine phosphokinase (CPK) ATP regeneration system demonstrated increased capture across all repair pathways as compared to a control. Nucleotide Excision Repair (NER) and mismatch repair (MMR) saw significant increases in repair activities not previously seen without the regeneration system. Temozolomide pretreated cells demonstrated increased repair activities in Direct Reversal repair of methylated adducts as compared to Hydrogen Peroxide pretreated cells and positive controls. Conclusions: The addition of an ATP regeneration system significantly increased capture of repair events by increasing local concentrations of ATP to support cellular enzyme activities. Increases in repair activates were seen across all pathways tested, indicating reproducible results. DNA damaging pretreatments indicated predictable up-regulations in certain DNA repair pathways matching the type of DNA damage event generated.

Citation Format: Lakotah Hardie, Amanda Richer, Jay Hesselberth. Multiplexed measurement of DNA repair capacity for tumor heterogeneity [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A092. doi:10.1158/1535-7163.TARG-19-A092
Abstract A108: Specific and traceless payload release from HER2-ADC containing LBG-linker prodrug by the action of lysosomal β-glucuronidase
Authors: Shin Dong Yeon , Jang Taeik , Song Ho Young , Kim Sung Min , Park Yun-Hee , Park Chang Sik , Oh Hwanhee , Baek Juyuel , Chae Jeiwook , Lee Chang-Sun
Published: March 2020

Many antibody-drug conjugates (ADCs) have actively been evaluated in clinical trials since Mylotarg was approved in 2001, but only 4 more have been approved by the FDA. The main reason for the poor results is yet to overcome worse risk-benefit than the expected from ADC approach. Controlled payload release specifically at cancer cells would significantly reduce systemic toxicity. Herein, we report that LBG000-linker can be selectively cleaved from ADC to release free toxins in a traceless manner by β-glucuronidase. Lysosomal β-glucuronidase is overexpressed in many cancer cell lines, and its enzymatic activity is much higher in acidic condition than neutral pH. As a model reaction, an LBG-linker-MMAE conjugate, LCB14-0302, was incubated with a β-glucuronidase in vitro resulting clean release of free MMAE while other lysosomal enzymes such as cathepsin B or heparanase did not generate the free toxin. We also confirmed that there was no free MMAE found when β-glucuronidase inhibitor, β-D-glucuronic acid, was treated. Also, lysosomal extract incubation with LCB14-0302 clearly demonstrated the traceless toxin release by β-glucuronidase regardless of other enzymatic disturbance. Therefore, these results strongly suggest that LBG-linker is highly stable and specific to β-glucuronidase among hundreds of enzymes in the lysosome.

Citation Format: Dong Yeon Shin, Taeik Jang, Ho Young Song, Sung Min Kim, Yun-Hee Park, Chang Sik Park, Hwanhee Oh, Juyuel Baek, Jeiwook Chae, Chang-Sun Lee. Specific and traceless payload release from HER2-ADC containing LBG-linker prodrug by the action of lysosomal β-glucuronidase [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A108. doi:10.1158/1535-7163.TARG-19-A108
Abstract A109: Repurposing low dose of quinoline methanol derivatives as a novel treatment for pediatric glioblastoma
Authors: Teng Jian , Carvalho Litia , Tabet Elie , Tannous Bakhos
Published: March 2020

Both primary brain tumors and secondary brain metastases that start in another part of the body and spreads to the brain, such as lung, breast, melanoma, kidney, nasopharynx, and colon cancers have limited or no response to chemotherapy. Through repurposing drug screen using pediatric glioblastoma cells established from newly diagnosed and recurrent patients, we identified that FDA-approved compound mefloquine have significant cancer killing properties. A growing body of evidence suggests that this extensively used anti-malaria drug may cause adverse neurological effects after crossing the blood-brain barrier and accumulates in the brain at relatively high concentration. To the amelioration of this problem, an analog campaign was conducted in selecting compounds lacking penetration but maintaining the anti-malarial properties using mefloquine as the leading compound. We therefore thought to take advantage of this unique characteristic and use low dose of these compounds and target the infiltrative disease reservoir while avoiding the neurological side effects. While this program focuses on high anti-malaria potency, low neurotoxicity (against rat neurons), and low brain absorption, we are interested in high anti-malaria potency, low general toxicity, and high brain/tumor concentration. Among these compounds, we selected TQM due to its promising results in efficacy, potency, and selectivity in killing brain tumor cells at a much lower range compared to mefloquine based on our quantitative structure-activity relationship model. Studies suggested that this group of drugs induce cell apoptosis in host and parasites through increased production of reactive oxygen species (ROS). Even though for a long time ROS is believed to promote cell proliferation; however, brain cancer cells, due to their high basal metabolic rate, are more susceptible to ROS. Inducing an increased intracellular ROS levels in brain cancer cells may trigger cell death and destroy malignant cells, while sparing normal cells. Our preliminary results confirmed that mefloquine and its derivatives induce apoptosis through autophagy and ROS signaling in brain cancer cells at a much lower concentration, and these compounds reduces tumor growth and extends animal survival in mouse models. Our results pave the way for translation of those compounds to the clinic.

Citation Format: Jian Teng, Litia Carvalho, Elie Tabet, Bakhos Tannous. Repurposing low dose of quinoline methanol derivatives as a novel treatment for pediatric glioblastoma [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A109. doi:10.1158/1535-7163.TARG-19-A109
Abstract A114: Quantitative mass spectrometry imaging: A game changer in the pharmaceutical industry
Authors: Ait-Belkacem Rima , Hochart Guillaume , Marini Joseph , Tomezyk Aurore , Mantefeul p , Jung B. , Bonnel David , Kenzie Don Mc , Stauber Jonathan
Published: March 2020

Developing a drug requires to demonstrate its efficacy in preclinical and clinical studies but also to characterize its pharmacokinetics properties. If Quantitative Whole-Body Autoradiography (QWBA) is still a gold standard to support the design of radiolabelled clinical studies, its lack of molecular specificity limits the understanding of the distribution of the drug and its metabolites, with no establishing of separate quantitative results for each target tissue of interest. On the contrary, the use of high resolution mass spectrometers such as FT-ICR MS allows a quantitative characterization of the metabolic profile of the drug at the tissue level in order to further understand its behaviour in the organism. A comparative analysis between QWBA and QMSI was applied as a complementary approach to follow chloroquine and one of its metabolites (desethyl-chloroquine) at different time-points after a single administration of a radiolabeled dose to Long-Evans male rats (30 mg/kg). 1H-Chloroquine and 1H-desethyl chloroquine were well detected by MALDI-FTICR in the eye including; the uveal tract, the vitreous humor, the lens and the Harderian gland. In the mid whole-body region, both compounds were also detected in various organs from the unique prepared T4h sections. Interestingly the two compounds were co-localized into the tissue sections and their distributions matched the zones obtained by QWBA. In QWBA additional regions containing the radiolabeled moiety could be clearly identified because of the high sensitivity of the technique. The advantage brought by MALDI QMSI was the ability to discriminate between parent drug 1H-choloroquine and its metabolite 1H-desethyl chloroquine so that each compound had its own distribution image and thus its own quantification data directly in one tissue section. The used labeled forms of both compounds during the matrix deposit allowed normalizing the data for each position targeted with the MALDI onto the section of interest and the calibration range of both 1H-choloroquine and 1H-desethyl chloroquine and quantifying each compound into the organs of interest with the ILC approach. Finally the QMSI was able to demonstrate the disappearance of the drug and its metabolite with time to better understand differential pharmacokinetics analysis demonstrating the additional input of the technology compared to QWBA.

Citation Format: Rima Ait-Belkacem, Guillaume Hochart, Joseph Marini, Aurore Tomezyk, p Mantefeul, B. Jung, David Bonnel, Don Mc Kenzie, Jonathan Stauber. Quantitative mass spectrometry imaging: A game changer in the pharmaceutical industry [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A114. doi:10.1158/1535-7163.TARG-19-A114
Abstract A129: Vertical inhibition overcomes adaptive resistance to KRASG12C inhibition
Authors: Ryan Meagan B , Cruz Ferran Fece de la , Phat Sarah , Myers David T , Shahzade Heather A , Hong Catriona B , Corcoran Ryan B
Published: March 2020

The three RAS genes are the most commonly mutated oncogenes in cancer and have long been considered an “undruggable” target. Alterations in KRAS occur at the highest frequency, with G12D, G12V, and G12C mutations being the most common. The recent development of novel covalent inhibitors selective for the KRASG12C mutation offers the unprecedented opportunity to target KRAS directly. However, prior efforts to target the RAS-MAPK pathway in have been hampered by adaptive feedback, which drives pathway reactivation and resistance. We find evidence of rapid adaptive RAS-MAPK pathway feedback reactivation following KRASG12C inhibition in the majority of KRASG12C models, including lung, colon, and pancreatic lines. Reactivation of the RAS-MAPK pathway correlates with activation of multiple upstream RTKs and subsequent increase of wild-type RAS activity. We find that co-inhibition of SHP2, a common node downstream of RTKs, or co-inhibition of individual RTKs such as EGFR or FGFR enhances the efficacy of KRASG12C inhibition both in vitro and in vivo by overcoming adaptive reactivation of the RAS-MAPK pathway. Notably, only the combination of KRASG12C and SHP2 inhibitor treatment displays tumor regressions in vivo in multiple models when compared to either inhibitor alone. We also see evidence that combining KRASG12C inhibitors with MEK or ERK inhibitors as an additional potential vertical inhibition strategy in KRASG12C models. We propose a vertical inhibition strategy anchored with KRASG12C inhibitors with either RTK or SHP2 inhibitors as a potential strategy for the treatment of KRASG12C mutant tumors.

Citation Format: Meagan B Ryan, Ferran Fece de la Cruz, Sarah Phat, David T Myers, Heather A Shahzade, Catriona B Hong, Ryan B Corcoran. Vertical inhibition overcomes adaptive resistance to KRASG12C inhibition [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A129. doi:10.1158/1535-7163.TARG-19-A129
Abstract A130: Cannabidiol overcomes oxaliplatin resistance by enhancing autophagy through reduction NOS3 and SOD2
Authors: Kim Bu Gyeom , Jeong Soyeon , Kim Dae Yeong , Kim Bo Ram , Kim Jung Lim , Park Seong Hye , Na Yoo Jin , Jo Min Jee , Yun Hye Kyeong , Jeong Yoon A , Lee Sun Il , Jeong Yoon A , Kim Hong Jun , Kim Han Do , Kim Dae Hyun , Lee Dae-Hee , Oh Sang Cheul
Published: March 2020

Although oxaliplatin is an effective chemotherapeutic drug for colorectal cancer (CRC) treatment, patients often develop resistance to it. Therefore, a new strategy for CRC treatment is needed. The purpose of this study was to determine the effect of cannabidiol (CBD), one of the components of the cannabis plant, in overcoming oxaliplatin resistance in CRC cells. We established oxaliplatin-resistant cell lines, DLD-1 R and colo205 R, in CRC DLD-1 and colo205 cells. Autophagic cell death was induced when oxaliplatin-resistant cells were treated with both oxaliplatin and CBD. Additionally, phosphorylation of nitric oxide synthase 3 (NOS3) was increased in oxaliplatin-resistant cells compared to that in parent cells. Combined treatment with oxaliplatin and CBD reduced phospho-NOS3 levels and NO production and resulted in the production of reactive oxygen species (ROS) by reducing the levels of superoxide dismutase 2, an antioxidant present in the mitochondria, causing mitochondrial dysfunction. Taken together, these results suggest that elevated phosphorylation of NOS3 is essential for oxaliplatin resistance. The combination of oxaliplatin and CBD decreased NOS3 phosphorylation, which resulted in autophagy, by inducing the overproduction of ROS through mitochondrial dysfunction, thus overcoming oxaliplatin resistance.

Citation Format: Bu Gyeom Kim, Soyeon Jeong, Dae Yeong Kim, Bo Ram Kim, Jung Lim Kim, Seong Hye Park, Yoo Jin Na, Min Jee Jo, Hye Kyeong Yun, Yoon A Jeong, Sun Il Lee, Yoon A Jeong, Hong Jun Kim, Han Do Kim, Dae Hyun Kim, Dae-Hee Lee, Sang Cheul Oh. Cannabidiol overcomes oxaliplatin resistance by enhancing autophagy through reduction NOS3 and SOD2 [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr A130. doi:10.1158/1535-7163.TARG-19-A130
Abstract B021: PARP16 is a novel target of talazoparib which contributes to synergy with adavosertib in SCLC
Authors: Palve Vinayak , Knezevic Clare , Luo Yunting , Li Xueli , Novakova Slivia , Welsh Eric , Fang Bin , Kinose Fumi , Haura Eric B , Monteiro Alvaro N , Koomen John M , Lawrence Harshani R , Rix Uwe
Published: March 2020

Background: Small cell lung cancer (SCLC) is a highly aggressive type of neuroendocrine tumor with a low survival rate, signifying an unmet medical need. Although PARP inhibitors (PARPis) display single-agent anticancer activity in SCLC, the underlying biology is poorly understood. In the present study, we determined the differential efficacy of multiple clinically relevant PARPis and identified a novel non-canonical and actionable PARPi target in SCLC. Methods: A panel of SCLC cell lines was analyzed for sensitivity to PARPis using the Cell Titer Glow (CTG) viability assay. Chemical proteomics and western blotting were performed to investigate the targets of PARPis. The effect of PARP16 on cell viability was analyzed using RNA interference. Drug combination screening for inhibition of viability was performed, network-wide signaling changes were determined using phosphoproteomics, and efficacy of drug combinations was determined in ex vivo PDX clonogenic assays. Results: Compared to other clinically relevant PARPis (rucaparib, olaparib, niraparib) talazoparib displayed the highest potency across a panel of SCLC cell lines, including cells negative for SLFN11, which has been suggested to indicate PARPi sensitivity. Subsequent chemical proteomics with SLFN11-negative SCLC cells identified the non-canonical PARP family member PARP16 as a unique talazoparib target, along with the eminent PARP1. Transient knockdown of PARP16 significantly reduced cell survival in certain SCLC cell lines, particularly in combination with olaparib. Furthermore, drug combination screens revealed synergistic activity of talazoparib when combined with the WEE1 inhibitor adavosertib in SCLC cells, which was more pronounced than the combination of adavosertib with olaparib, which has no activity against PARP16. Silencing of PARP16, in combination with olaparib and adavosertib, with enhanced cell killing, and global phosphoproteomics identified network-wide signaling crosstalk inhibition by the talazoparib/adavosertib drug combination. Conclusion: Our data suggest that in addition to the well-studied PARP1 targeting mechanism, PARP16, a non-canonical member of the PARP family, is likely involved in talazoparib’s overall mechanism of action and may constitute a novel actionable target in SCLC.

Citation Format: Vinayak Palve, Clare Knezevic, Yunting Luo, Xueli Li, Slivia Novakova, Eric Welsh, Bin Fang, Fumi Kinose, Eric B Haura, Alvaro N Monteiro, John M Koomen, Harshani R Lawrence, Uwe Rix. PARP16 is a novel target of talazoparib which contributes to synergy with adavosertib in SCLC [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B021. doi:10.1158/1535-7163.TARG-19-B021
Abstract B093: 50 gene breast cancer (BC) RNA subtype classifier and colorectal cancer (CRC) CMS subtype classifier applied to 90 prostate cancer (PC) patients reveals distinct subtype differences
Authors: Adashek Jacob J , Pal Sumanta K , Veerapaneni Saihitha , Obeid Elias , Szeto Christopher W , Reddy Sandeep K , Nussenzveig Roberto , Agarwal Neeraj
Published: March 2020

Background: BC and CRC are heterogeneous diseases with several distinct disease subtypes. A 50-gene qPCR assay (PAM50) identifies 5 intrinsic biological subtypes: luminal A, luminal B, HER2-enriched, basal-like, and normal-like in breast cancer. Consensus molecular subtype in CRC (CMS) identifies 4 subtypes: CMS1-4. PC can be classified by specific genomic rearrangements, such as the TMPRSS2-ERG fusion occur or whether specific mutations such as SPOP and FOXA1 or germline BRCA is present, yet 25% of PC remains difficult to classify based on molecular drivers. We used comprehensive genomic and transcriptomic data to determine the association of somatic and germline mutations in molecular PC subtypes using the CMS and 50-gene breast cancer classifiers. Methods: Retrospective analysis on Whole exome (WES) DNA tumor and paired germline and matched deep whole transcriptomic sequencing (RNA-Seq) (∼200x106reads per tumor) data from NantHealth was performed. BC Intrinsic Subtypes and CMS sorting based on RNAseq assay was used to classify PC into 5 BC subtypes and CMS 1-4. Results: 90 PC patients were classifiable using RNAseq and WES DNA. BC subtype distribution was 64.4% Luminal A, 31.1% Luminal B, 2.2% HER2-enriched, 2.2% normal-like, and 0 classified as Basal-like. The CMS subtype distribution was 83.3% CMS4, 15.6% CM1, 1.1% CMS3, and 0 CMS2. Overlap comparison revealed that almost all LumA are CMS4 (56/58, 96.6%) and CMS1 are LumB (11/14, 78.6%). Analysis of germline DDR mutations in LumB compared to LumA showed LumB has significantly more DDR mutants than LumA (OR 2.68, p = 0.035 one-sided Fishers exact test). Conclusions: PC molecularly profiled using 50-gene and CMS classification we found LumA subtype as the predominant subgroup characterized by low TMB and high CMS4. LumB was characterized as higher TMB and greater CMS1 suggesting potential immunotherapy eligibility in this subtype. The presence of potentially pathogenic germline variants in DDR genes is associated with subsequent somatic increase in TMB, however, somatic expression subtyping (by BC or CRC types) is a stronger indicator of TMB than germline pPV in DDR genes.

Citation Format: Jacob J Adashek, Sumanta K Pal, Saihitha Veerapaneni, Elias Obeid, Christopher W Szeto, Sandeep K Reddy, Roberto Nussenzveig, Neeraj Agarwal. 50 gene breast cancer (BC) RNA subtype classifier and colorectal cancer (CRC) CMS subtype classifier applied to 90 prostate cancer (PC) patients reveals distinct subtype differences [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B093. doi:10.1158/1535-7163.TARG-19-B093