bluuebunny/crossref_metadata_2025 huggingface dataset:
Mechanistic study of the superior anti-cancer properties of a first-in-class small molecule targeting PCNA.
Authors: Gu Long , Lingeman Robert , Li Min , Hickey Robert , Liu Yilun , Malkas Linda
Published: May 2019

e14636

Background: Proliferating cell nuclear antigen (PCNA), through its interaction with various proteins involved in DNA synthesis, cell cycle regulation, and DNA repair, plays a central role in maintaining genome stability. We previously reported a novel cancer associated PCNA isoform (dubbed caPCNA), which was predominantly expressed in a broad range of cancer cells and tumor tissues, but not significantly in non-malignant cells. We found that the caPCNA-specific antigenic site lies between L126 and Y133, a region within the interdomain connector loop of PCNA that is known to be a major binding site for many of PCNA’s interacting proteins. A cell permeable peptide harboring the L126-Y133 sequence inhibited PCNA function in cancer cells and selectively kills cancer cells and xenograft tumors. Methods: Based on these observations, we sought small molecules targeting this peptide region of PCNA as potential broad-spectrum anti-cancer agents. Results: Our effort led to a drug candidate, AOH1996, which selectively kills a broad range of cancer cells at high nanomolar concentrations, but is not associated with significant toxicity to non-malignant cells. It also works synergistically with DNA damaging chemotherapeutic drugs, such as cisplatin and irinotecan, to selectively kill cancer cells. This compound is orally available to animals and suppresses tumor growth in a dosage form compatible to clinical applications. Importantly, it doesn’t cause significant toxicity at 2.5 times its effective dose. Mechanistically, AOH1996 competes with T3, a known PCNA ligand, for binding to PCNA. However, the mechanism by which AOH1996 exerts its effect on cancer cells may not be identical to what have been reported for the T3 analogs. In particular, we found that AOH1996 interferes with the association of PCNA and MCM7 to euchromatin, leading to DNA replication stress, blockade of homologous recombination-mediated DNA repair, and induction of apoptosis in cancer cells. Conclusions: These findings demonstrated the potential of this compound as a novel therapeutic agent warranting clinical investigation for cancer treatment. We have started planning a phase 1 clinical study for this compound.
NCI9782: A phase 1 study of talazoparib in combination with carboplatin and paclitaxel in patients with advanced solid tumors.
Authors: Turk Anita Ahmed , Leal Ticiana , Chan Nancy , Wesolowski Robert , Spencer Kristen Renee , Malhotra Jyoti , Lang Joshua Michael , McNeel Douglas G. , O'Regan Ruth , Mehnert Janice M. , Eickhoff Jens C. , Liu Glenn , Wisinski Kari Braun
Published: May 2019

e14640

Background: Poly(ADP-ribose) polymerase (PARP) enzymes are involved in DNA repair and activated by DNA strand breaks. DNA damage from carboplatin is associated with activation of PARP. Preclinical data indicate that PARP inhibition and trapping potentiates the anti-tumor effect of platinum chemotherapy. Talazoparib (T) is an oral, selective PARP inhibitor. This phase I study combines T with carboplatin (C) and paclitaxel (P). Methods: Two dosing schedules are being investigated. C is administered on day 1 and P on days 1, 8, and 15 of a 21-day cycle. T (100-1000mcg) is dosed once daily for days 1-7 (schedule A) or days 1-3 (schedule B). Dose escalation is by 3+3 design. Patients (pts) must have tumor type that is expected to respond to C + P or have BRCA germline or somatic mutation and adequate organ function. After 4-6 cycles of combination therapy, pts may continue the combination, change to C and intermittent T without P or change to T alone. Each schedule will have a 6 pt dose expansion at the MTD. The dose level (DL) 1 for schedule B is the previously reported MTD from schedule A (T 250mcg with C AUC 6 + P 80mg/m2). Results: Schedule B results are reported: 15 pts (median age 56 yrs [range 43-76]) have been enrolled. Primary malignancies include colorectal (4), pancreas (4), prostate (2), urothelial (2), and other (3). Dose was initiated at Schedule A MTD. DL2 (T 350mcg with C AUC 6 + P 80mg/m2) exceeded the MTD with 2 of 6 pts experiencing hematologic dose limiting toxicities (DLTs). DL1 is the confirmed schedule B MTD. Dose expansion to 6 pts is ongoing. Of the 11 pts with measurable disease, 3 (27%) had PR and 5 (45%) had SD. Pts were on study a median of 10 weeks (range 5-36+). Most common grade 3/4 AEs include leukopenia (53%), neutropenia (47%), and anemia (47%). One grade 5 AE of intracranial hemorrhage occurred, possibly related to therapy in the setting of grade 3 thrombocytopenia and concern for CNS disease. Conclusions: The schedule B MTD and RP2D is T 250 mcg with C AUC 6 and P 80mg/m2. Data from the full dose expansion will be presented. This combination was tolerated with prolonged responses seen at lower dose T in combination with C+P. Clinical trial information: NCT02317874.
Treatment of advanced gastrointestinal cancer with genetically modified autologous mesenchymal stem cells: Final results of the phase 1/2 TREAT-ME-1 trial.
Authors: von Einem Jobst C. , Guenther Christine , Volk Hans Dieter , Gruetz Gerald , Hirsch Daniela , Salat Christoph , Stoetzer Oliver J , Nelson Peter J , Michl Marlies , Modest Dominik Paul , Holch Julian , Angele Martin K , Bruns Christiane J. , Niess Hanno , Heinemann Volker
Published: May 2019

e14648

Background: TREAT-ME-1, a Phase 1/2 open-label, first-in-human, first-in-class trial, evaluated the safety, tolerability and efficacy of treatment with genetically modified autologous mesenchymal stromal cells (MSC),MSC_ apceth_101, in combination with ganciclovir in patients with advanced gastrointestinal adenocarcinoma. Methods: All patients (6 in Phase 1; 7 in Phase 2) received three treatment cycles of MSC_apceth_101 on Day 0, 7, and 14 followed by ganciclovir administration according to the manufacturer’s instructions for 48─72 hours after MSC_apceth_101 injection. Three patients were treated with a total dose of 1.5 x 106cells/kg MSC_apceth_101 (DL1). Ten patients were treated with a total dose of 3.0 x 106cells/kg MSC_apceth_101 (DL2). Immunological and inflammatory markers were assessed.Time to progression, overall survival was evaluated by RECISTwithin 56 days after administration of MSC_apceth_101 administration. Results: A total of 54 AEs occurred in 9/13 patients, thereof 18 AEs in three patients of DL1 and 36 AEs in six patients of DL2. Six AEs were CTC-AE grade 3, all other AEs were lower CTC-AE grades.Five SAEs emerged in two patients of DL1 and six SAEs in four patients of DL2. Eight patients achieved stable disease (change in target lesions of -2 to +28%). For all patients, the median time to progression was 1.9 months (95% CI: 1.3, 3.6 months). Median overall survival could not be estimated as 8/10 patients were still alive at the end of the study (one year) and therefore censored. Post-study observation of patients showed a median overall survival of 15.6 months (ranging from 2.2─27.0 months). Treatment with MSC_apceth_101 and ganciclovir did not induce a consistent increase or decrease in levels of any of the tumor markers analyzed. No clear trends in the immunological markers assessed were observed. Conclusions: MSC_apceth_101 in combination with ganciclovir was safe and tolerable in patients with advanced gastrointestinal adenocarcinoma, with preliminary signs of efficacy in terms of clinical stabilization of disease. Clinical trial information: NCT02008539.
Identification of new targets and biomarkers in lung cancer.
Authors: Papasotiriou Ioannis , Parsonidis Panagiotis , Ntanovasilis Dimitrios-Athanasios , Iliopoulos Aggelos C , Beis Georgios , Apostolou Panagiotis
Published: May 2019

e14656

Background: Lung cancer is the second most common type of cancer in both genders; however is the leading cause of cancer deaths. Adenocarcinoma constitutes more than 60% of non-small cell lung cancer (NSCLC), and accounts for approximately 40% of lung cancers. Therefore, early prediction, prognosis of the disease and treatment strategy is of primary importance. Molecular biology assays, including microarrays, enable identification of new targets. The above targets can be used either as biomarkers, useful for early prediction and prognosis of the disease, or as drugable targets, upon proof of their biological action. The present study aimed to identify new biomarkers and/or targets in lung cancer. Methods: Whole genome gene expression microarrays have been performed for 12 lung cancer cell lines (provided be ECACC, ATCC), representing non-small cell lung (adenocarcinoma, squamous, large cell) and small cell lung cancer. Real-time experiments were followed for validation of thirty genes that were overexpressed. The experiments were performed including other cell lines, representing breast, colon, prostate, hepatocellular, melanoma and pancreatic cancer. Sample from healthy donors, as well as normal lung epithelial cell were used. Results: Among the genes that were studied, there was observed downregulation or none expression in normal donors as well as in normal lung epithelia cells. VSIG10 overexpressed in adenocarcinoma and large cells, while no expression observed in breast, prostate, melanoma and pancreatic cancer. DCBLD2 overexpressed in all NSCLC samples, but not in small cells. LIFR was overexpressed mainly in squamous cells, while no expression or downregulation observed in the other types of cancer. The same expression profile was mentioned for PDGFC. Conclusions: Taking everything into consideration, it is obvious that the identification of new biomarkers and/or targets for lung cancer is of primary importance. The present study revealed different genes, with different biological function that might be used primary as biomarkers. Their role need to be clarified also in clinical samples as to be used as drugable targets. However, the above data are encouraging for identification of new therapeutic models in lung cancer.
Integrating CRISPR screening with tumor genomic analyses to identify therapeutic targets in hepatocellular carcinomas (HCC) independent of P53.
Authors: Sheel Ankur , Kwan SuetYan , Xue Wen
Published: May 2019

e14659

Background: Hepatocellular carcinoma (HCC) is an aggressive subtype of liver cancer with few effective treatments. Moreover, the underlying mechanisms that drive HCC pathogenesis remain poorly characterized. Identifying genes and pathways essential for HCC cell growth will aid the development of new targeted therapies for HCC. Furthermore, the P53 pathway is frequently mutated in HCC therefore identifying targets with therapeutic efficacy irrespective of P53 status is warranted. Methods: To identify kinases essential for HCC proliferation, we performed a kinome wide CRISPR screen in human HCC cell lines with varying P53 mutations and validated our findings using CRISPR-Cas9 mediated genetic manipulations in human HCC cell lines in-vitro and in-vivo. Furthermore, we performed an integrated cancer genomics analyses using patient data from TCGA and the NCI to validate the relevancy of our findings. Results: We identified transformation/transcription domain-associated protein (TRRAP) as an essential gene for HCC cell proliferation. we show that depletion of TRRAP or its co-factor, histone acetyltransferase KAT5, inhibits HCC cell growth via induction of P53, P21 and RB-independent senescence in-vitro and in-vivo. Furthermore, we find that TRRAP is upregulated in HCC patient samples independent of TP53 mutations. Integrated cancer genomics analyses using both HCC patient data derived from TCGA and from RNA-sequencing of our in-vitro model identified a chromosomal instability signature that was regulated by TRRAP/KAT5 in-vitro. Furthermore this chromosomal instability signature was also upregulated in HCC patients. Finally, we identify TOP2A as a target in this pathway as genetic depletion of TOP2A inhibited cell growth via induction of senescence. Conclusions: Our results uncover a role for TRRAP/KAT5 in promoting HCC cell proliferation via activation of mitotic genes in order to potentiate a chromosomal instability signature. Our findings suggest that targeting the TRRAP/KAT5 complex and TOP2A is a therapeutic strategy for HCC, even in tumors that have escaped P53 and RB tumor suppressive programs.
Implication of DYRK1B kinase in dormant glioblastoma cancers and utilization of DYRK1B inhibitors as a novel therapeutic strategy for glioblastoma.
Authors: Vilenchik Maria , Kuznetsova Alexandra , Frid Michael , Duey Menelik , Damiani Arianna , De Leon Lita , Law Jason , Golbin Denis A. , Shishkina Ludmila V. , Potapova Olga
Published: May 2019

e14670

Background: A major determinant of cancer resistance and recurrence is the presence of quiescent cancer cells. It has long been known that quiescent cancers are resistant to a wide variety of anti-cancer therapeutics relative to actively proliferating cells. However, the presence and relative size of the quiescent compartment is not revealed by standard H&E staining and morphological tumor analysis and, therefore, the data demonstrating its importance and prevalence by tumor type are lacking. DYRK1B kinase (serine/threonine-protein kinase dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 1B) is associated with growth and survival of many types of cancer cells and was shown to play a key role in maintaining the cancer cells in reversible quiescent state by stabilizing the CDK inhibitor p27 and by inducing the breakdown of cyclin D isoforms. Methods: We conducted an immunohistochemical study of 80 human glioblastoma (GBM) tumors (primary / treatment-naive, Grade IV) and 10 cases of normal postmortem brain tissues for expression of DYRK1B, and quiescence and proliferation markers p27 and Ki-67, respectively. For subsection of these tissues, high levels of DYRK1B and p27 expression were observed. Results: In this study, we investigated the functional and therapeutic relevance of DYRK1B expression in glioblastomas, the most common brain cancer in adults. It is an aggressive, highly resistant to treatment tumor type with median survival time of less than 16 months following surgery and treatment. Conclusions: There was a statistically significant correlation between immunostaining of DYRK1B and p27, indicating the presence of dormant cancer cells mixed with the proliferating cancer cells within the patients' tumors. We suggest that DYRK1B inhibitors in combination with chemotherapeutics agents have a potential to improve patient treatment outcome and prolong survival.
Role of ultrasound contrast agent in a tertiary cancer center: Experience in 75 patients with liver biopsy.
Authors: Rahnemai-Azar Amir Ata , Kikano Elias , Smith Daniel Arnold , Ramaiya Nikhil H. , Azar Nami R
Published: May 2019

e14671

Background: The sensitivity of B-mode ultrasound (US) in detecting hepatic lesions has been reported to be between 55-85%. A recently FDA approved US contrast agent has been shown to increase the detectability of liver lesions; yet, there is paucity of knowledge regarding its application in US guided biopsies. The aim of this study was to determine accuracy and applicability of contrast enhanced US (CEUS) guided biopsies of liver lesions in cancer patients. Methods: A prospective registry of 75 patients (M = 54%, F = 46%, Age: 63y±14), that underwent CEUS guided liver biopsy in 2017-2018, were retrospectively reviewed. The decision to apply US contrast was made based on pre-procedural and intra-procedural clinical findings. Lesions with negative biopsy results were reviewed in follow up imaging to determine any evidence of potential positive malignancy. Results: History of renal failure that precluded application of iodinated IV contrast was the main pre-procedural indication (17%). Intra-procedural indications included: poor visualization of lesions in B-mode US (78%), targeting enhancing part of lesions (necrotic, post ablation or cystic) (16%), and lesion selection (6%). Diameter of the lesions were ≤ 20mm in 46% of patients. Biopsy results were positive in 50 patients, and included 32% primary liver tumors (hepatocellular carcinoma and cholangiocarcinoma) and 68% metastasis (pancreas: 24%, breast: 12%, GI tract: 10%, lung: 8% and other: 14%). Of the 25 patients with negative biopsy results, 6 did not have follow up imaging and 3 were later found to have positive malignancy (Negative predictive value: 84%, accuracy: 96%). Genomic sequencing was established in 34% of the patients. Based on the biopsy/genomics results, patients underwent systemic treatment (56%), locoregional treatment (16%), combination (6%) or surgery (2%). 10% of patients selected hospice care and 10% lost follow up. Conclusions: CEUS increases the accuracy of US guided biopsies of focal liver lesions, by improving visualization, targeting viable parts and also in patients with renal failure, or small lesions. Findings from these biopsies could improve management of cancer patients by categorizing them into different treatment groups.
A retrospective analysis of venous thromboembolism trends in chemotherapy-induced anemia: Red blood cell transfusion versus erythrocyte stimulating agent (ESA) administration.
Authors: Bryer Emily Jane , Henry David H.
Published: May 2019

e14685

Background: Anemia is a common and unfortunate consequence of chemotherapy; patients receiving a variety of chemotherapy regimens often develop chemotherapy–induced anemia (CIA), which contributes to poor outcomes including increased mortality. Prompt and effective treatment of CIA is essential to prevent fewer chemotherapy dose delays and reductions. Optimal therapy of CIA is controversial and involves the solitary and combined use of intravenous iron, red blood cell (RBC) transfusions, and erythropoietin stimulating agents (ESAs). Despite the baseline coagulopathies present in patients with malignancy, administration of both RBC transfusions and ESAs is associated with venous thromboembolism (VTE). It remains unknown whether the risk of VTE in patients with CIA is greater among patients who receive RBC transfusions or ESAs. Methods: A retrospective single-institution study analyzed 7360 patients with varying malignancies who developed CIA and received ESAs and RBC transfusion from 1998-2017. These patients were evaluated for subsequent development of VTE and categorized by prior receipt of RBC transfusion or ESA. Results: Among the 7360 patients with CIA, 5503 received either RBC transfusion or ESA and 1857 received both. Among all patients, 3466/7360 (47.1%) developed a VTE. The absolute risk of developing a VTE with receipt of a RBC transfusion was 0.38 compared to 0.19 with ESA. Patients with CIA who received RBC had twice the risk of developing a VTE compared with those who received ESA (p < 0.0001). Conclusions: While both RBC transfusion and ESA administration are independently associated with VTE, our data suggests a greater risk of VTE development with RBC transfusion as compared with ESA administration.[Table: see text]
Biofabricated tumor microenvironments for studying colorectal cancer in vitro and in vivo.
Authors: Devarasetty Mahesh , Herberg Samuel , Dominijanni Anthony , Willey-Shelkey Ethan , Skardal Aleksander , Soker Shay
Published: May 2019

e14689

Background: Microenvironmental mechanics have a tremendous effect on the progression, phenotype, and therapeutic response of cancer cells positioning it as a high-potential target for novel therapeutic development. Laboratory modeling of the microenvironment and its multitude of effects is imperative for developing new avenues of anti-cancer therapy that can target non-traditional vectors such as the extracellular matrix (ECM) and stromal cells. Researchers have developed in vitro models of the tumor microenvironment (TME) to meet this need. While in vitro modeling is an important step in therapeutic development, there are few studies that validate in vitro generated results to gold-standard in vivo models, and further, to patient-derived data. Previously, we have developed a model of the colorectal tumor microenvironment and found a connection between collagen ultrastructure and cancer cell phenotype. Using this characterized organoid model, we implant bioengineered TMEs into mice to track long-term growth and progression of cancer and compare our results to clinical biopsies. Methods: Tumor organoids are produced by combining stromal cells and type I collagen. Cancer cell spheroids are embedded into the organoid for long term observation. Organoids are either observed in vitro or implanted subcutaneously into mice for in vivo tracking. Results: Organoids retain structure and viability during long term culture in vitro and in vivo, and embedded cancer cells respond significantly differently depending on the architecture of the surrounding TME. Cancer cells assume a mesenchymal, invasive, and proliferative phenotype in unorganized TMEs, and revert to an epithelial phenotype in an ordered TME. In addition, analysis of biopsied tissue, across tumor grade, demonstrates a correlation between cancer cell phenotype and microenvironmental architecture. Conclusions: In all this is the first study to establish a connection between TME micro-structure and cancer cell phenotype consistently across three distinct research modalities. These results have the potential to pave the way for utilizing bioengineered microenvironmental models as therapeutic development platforms and for targeting TME micro-structure to control colorectal cancer cell progression.
Anticoagulants for the treatment of venous thromboembolism in patients with cancer: An updated pairwise and network meta-analysis of randomized trials.
Authors: Sidahmed Shima , Abdalla Ahmed , Kheiri Babikir , Bala Areeg , Salih Mohammed , Kafri Zyad , Bachuwa Ghassan
Published: May 2019

e14692

Background: Cancer-associated venous thromboembolism (VTE) is common. Although low molecular weight heparin (LMWH) is the standard therapy in this setting, little is known with regard to non-vitamin K antagonist oral anticoagulants (NOACs). Therefore, we thought about evaluating the safety and efficacy of various anticoagulants in this vulnerable population. Methods: Electronic database search was conducted to identify randomized clinical trials (RCTs) that compared LMWH, NOACs, and/or vitamin-K-antagonists (VKA) in cancer patients. We performed frequentist direct and Bayesian network meta-analysis using random-effects model to calculate odds ratios (ORs), 95% confidence intervals (CIs), and 95% credible intervals (CrIs). The primary outcome was VTE (pulmonary embolism and deep-vein thrombosis) recurrence. Secondary outcomes were major bleeding and all-cause mortality. Results: We identified 13 RCTs with 6,595 total patients (mean age 62.4 ± 12.2; 50.4% female; 17.7% hematological malignancies; and 6 months median follow-up). The most common cancer type was colorectal and 48% of the population had metastatic cancer at baseline. NOACs were associated with significantly reduced VTE recurrence compared with VKA (OR = 0.58; 95% CI = 0.40-0.83; P < 0.01; number needed to treat [NNT] = 40) and LMWH (OR = 0.46; 95% CI = 0.25-0.85; P = 0.01; NNT = 20). LMHW was associated with significantly reduced VTE recurrence compared with VKA (OR = 0.52; 95% CI = 0.39-0.71; P < 0.01; NNT = 18). NOACs were associated with significantly reduced major bleeding compared with VKA (OR = 0.56; 95% CI = 0.35-0.91; P = 0.02; NNT = 64). There was no significant difference identified between the anticoagulant groups in regard to all-cause mortality. Conclusions: Among cancer patients with VTE, NOACs were associated with significantly reduced VTE recurrence compared to LMWH and VKA, and significantly reduced major bleeding compared with VKA. LMWH was associated with significantly reduced VTE recurrence compared with VKA.
A phase I study investigating the absorption, metabolism, and excretion of [14C] anlotinib in patients with advanced refractory solid tumors.
Authors: Liu Yiqian , Xu Tongpeng , Liu Lianke , Shu Yongqian
Published: May 2019

e14701

Background: Anlotinib is a novel oral multi-targeted receptor tyrosine kinase inhibitor, which selectively inhibits VEGFR2/3, FGFR1-4, PDGFR α/β, c-Kit, and Ret, and has shown antitumor activity in patients with advanced refractory solid tumors. The human plasma pharmacokinetics of anlotinib have been described previously, but detailed analyses of the absorption, metabolism and excretion pathways of anlotinib have not yet been published. Methods: Six male patients were divided into two groups, one group included two patients was performed by 13.51mg Anlotinib hydrochloride and 0.64 mg/80 μCi [14C] anlotinib hydrochloride and the rest four patients in another group received 13.19mg anlotinib hydrochloride and 0.96mg/120 μCi [14C] anlotinib hydrochloride, followed by collection of blood, urine, and fecal samples. The primary outcomes of study were absorption, metabolism, and excretion of [14C] anlotinib and the second outcomes were efficacy and safety. Results: In plasma, the average time of total plasma radioactivity peak (Tmax) was 4.42h and the average peak concentration (Cmax) was 18.80 ng Eq./g. The average value of AUC last was 4071 h∙ng Eq./g, AUC0-∞ was 13555 h∙ng Eq./g, and MRT0-t was 145h. The average recovery rate of total radioactivity in urine and feces was 62.03%. Prototype drugs and metabolites were mainly excreted through feces, accounting for 48.52% of the administered dose, otherwise rine excretion accounted for 13.51%. The liver and kidney were the main drug excretion organs. Five patients developed adverse reactions, and one person developed SAE. Three of study population were assessed as stable disease (SD), two with partial response (PR), and one had progressive disease (PD). Conclusions: Anlotinib showed a favourable pharmacokinetic profile with high absolute bioavailability and an extensive hepatic metabolism. The adverse events and efficacy were as expected. Clinical trial information: CTR20160329.
Pharmacokinetics of ciclopirox prodrug, a novel agent for the treatment of bladder cancer, in animals and humans.
Authors: Weir Scott James , Wood Robyn , Baltezor Michael Jay , Reed Greg , Brinker Amanda E , Ham Tammy , Schorno Karl , Toren Paul , Ramamoorthy Prabhu , Zhukova-Harrill Valentina , Dalton Michael , McCulloch William , Patel Manish R. , Ulahannan Susanna Varkey , Burris Howard A. , Falchook Gerald Steven , Jensen Roy A. , Anant Shrikant , Taylor John Arthur
Published: May 2019

e14705

Background: Ciclopirox Prodrug (CPX-POM) is a novel anticancer agent currently being evaluated in patients with advanced solid tumors participating in a First-in-Human, Phase 1 safety, dose tolerance, pharmacokinetics (PK) and pharmacodynamics trial at four US sites. In vitro and in vivo preclinical proof of principle was established in high grade human urothelial cancer cell lines as well as a mouse model of bladder cancer.Methods: A series of in vivo PK studies were conducted in mice, rats and dogs to characterize the absolute bioavailability of CPX following intravenous (IV), subcutaneous (SC) and oral administration of CPX-POM. The single dose and steady-state plasma and urine pharmacokinetics of CPX-POM are also currently being characterized in patients participating in the ongoing Phase 1 trial. Plasma and urine concentrations of the prodrug and metabolites were determined by LC-MS/MS validated in each specie and matrix. Non-parametric pharmacokinetic parameters were generated from resultant plasma and urine drug and metabolite concentration-time data. Results: CPX-POM is rapidly and completely metabolized to CPX in blood via circulating phosphatases in animals and humans. CPX is completely bioavailable following IV CPX-POM administration in mice, rats and dogs. CPX and its major inactive glucuronide metabolite (CPX-G) are extensively eliminated in urine in all animal species. SC administration of CPX-POM demonstrated excellent bioavailability in rats and dogs. Following IV administration of 30-900 mg/m2CPX-POM to patients, the apparent elimination half-life of CPX ranged from 2 to 8 hours, CPX systemic exposure was dose-proportional and time-independent in cancer patients, and a major portion of the dose was eliminated as CPX-G. Conclusions: IV CPX-POM achieves plasma and urine CPX exposures that exceed in vitro IC50 values several-fold at well tolerated doses in animals and humans. CPX pharmacokinetics observed in animals were predictive of human systemic clearance based on allometric scaling. Clinical trial information: NCT03348514.
Effect of MALAT1 in the crosstalk between nucleus and mitochondria on mitochondrial reprogramming in hepatocellular carcinoma cells.
Authors: Zhao Yijing , Cui Jiuwei , Hu Jifan , Hoffman Andrew R
Published: May 2019

e14711

Background: Mitochondria-nuclear crosstalk is a bidirectional pathway of communication between mitochondria and nucleus that influences many cellular and organismal activities. This crosstalk can regulate several oncogenic pathways involved in tumorigenesis. MALAT1 (metastasis-associated lung adenocarcinoma transcript 1), a nucleus-encoded lncRNA, dysregulated in multiple human malignancies is recently found to drive mitochondria dysfunction in Mitochondria-nuclear crosstalk. Methods: RNA sequencing, “RNA reverse transcription-associated trap sequencing” (RAT-seq), RNA immunoprecipitation(RIP), fluorescence in situ hybridization (FISH), were performed to detect the position of the MALAT1, and its interaction protein and DNAs in HepG2 cell line. After silencing MALAT1 by shRNAs, Seahorse, ATP production, lysotracker staining, Western blot, and electronic microscope were used to measure metabolism, ROS amount, mitophagy, apoptosis and mitochondrial morphology of the silencing cell lines. Results: By combining mitochondrial RNA-Seq with FISH, it is surprised to discover that MALAT1 was enriched in the mitochondria of HepG2 cells. Using RAT-seq approach, MALAT1 was found to utilized it 3’-fragment to interact with multiple loci of mitochondrial DNA( D-loop, COX2, ND3, and CYTB ). The RIP and affinity RNA pulldown assays suggested that the RNA-binding protein HuR mediated the transportation of MALAT1 to the mitochondria. Also, mitochondria transmembrane protein mitochondrial carrier 2(MTCH2) is found to interacted MALAT1, suggesting that MALAT1 may through the MTCH2 to get into the inside of the mitochondria. Knockdown of MALAT1 induced multiple abnormalities in mitochondrial functions, including low OXPHOS, low ATP production, reduced mitophagy, declined mtDNA copy number, and activation of the mitochondrial apoptosis pathway. Conclusions: Together, this study greatly expands our knowledge of the nucleus-encoded lncRNA MALAT1 driving the mitochondria dysfunction. Our study establishes MALAT1 as a regulator through interacting with MT-DNA, HuR, MTCH2 protein, revealing a new regulatory mechanism of mitochondria. Many novel nucleus-encoded RNAs existing in mitochondria were identified at the first time, suggesting novel biological functions of lncRNAs, laying the foundation for further clarifying their roles.
Effect of tivantinib on VEGF signaling pathways and apoptosis of gastric cancer cells with c-MET or VEGFA amplification.
Authors: Kim Hyeong Su , Zang Dae Young , Sohn Sung-Hwa , Kim Bohyun , Sul Hee Jung
Published: May 2019

e14719

Background: VEGFA is the key mediator of angiogenesis in cancer and previous studies reported that VEGFA expression was significantly up-regulated in gastric cancer tissues compared with matched normal tissues. We showed that increased levels of VEGFA are significantly associated with expression of hepatocyte growth factor receptor (MET) (r = 0.6255, P < 0.0001). In addition, it is well known that c-MET is potentially a highly plausible target for cancer therapy in gastric cancer. In this study, cytotoxic activity of tivantinib were evaluated in gastric cancer cells with high c-MET expression or VEGFA amplification. Methods: In this study, Western blot and quantitative real-time PCR analysis were used to detect the expression of protein and genes after treatment of tivantinib. In addition, MTS, flow cytometry, and migration assay were used. Results: Tivantinib inhibited growths of a high c-MET-expressed or VEGFA-amplified cell lines. Furthermore, in migration and apoptosis analysis, tivantinib induced apoptosis of SNU620, MKN45 (carried VEGFB mutation), AGS, and MKN28 cells but not in KATO III (carried VEGFB and VEGFC mutation) cells. We also found that tivantinib inhibited the VEGF signaling pathway and MYC expression in VEGFA-amplified gastric cancer cell lines. Conclusions: The data indicate that tivantinib could be a potential therapeutic agent for the treatment of gastric cancer with high c-MET expression or VEGFA amplification.
FBXO4 as a novel ubiquitin ligase that targets Cyclin D in the pathogenesis of breast cancer.
Authors: Dhar Ruby , Rath Goura Kishor , Karmakar Subhradip
Published: May 2019

e14722

Background: Members of ubiquitin ligase family are reported to be associated with cancer. Genome wide RNAi screen using Dharmacon on target Plus RNAi library identified FBXO4 as one of the potential hit . We therefore aimed to study its detailed role on proliferation of MCF7 breast cancer cells with special focus on cell cycle genes. We want to expand it further and study its involvement in context to E2 signaling in breast cancer. We hypothesized that increase in CCND1 expression, could mimic some of the E2 transcriptional regulation and that CCND1 overexpression could lead to E2 independent ER alpha binding to estrogen response element (ERE). Methods: Cell lines: MCF7 grown in MEM alpha supplemented with 10% FBS . For stimulation studies, cells were maintained in phenol free MEM alpha supplemented with 10% FBS and 10 nM estradiol. RNAi knockdown studies: On-target plus siRNA were used to knockdown FBXO4 in MCF7 cells. Western Blot: Western Blot was performed following standard protocols. IHC/Tissue Array: Paraffin embedded malignant breast cancer (ER/PR/EGFR +ve) tissue samples that are scored by pathologist were used for the study. Gene Expression Studies: Microarray and NGS based approach. Results: FBXO4 has been identified as a potential hit in our RNAi screen showing a hyper-proliferative phenotype in MCF7 cellsupon its knockdown. Western blot and ubiquitination assay identified cyclin D (CCND1) as a dedicated FBXO4 target as revealed by confocal microscopy where FBXO4 knockdown showed an enrichment of CCND1 in cytoplasm. Ingenuity pathway analysis and ChIP assay revealed an exploitation of E2 signaling and enhanced recruitment of ER alpha to estrogen response element (ERE) in FBXO4 depleted cells. CCND1 further seems to co-occupy ESR binding motifs thereby representing ER functions. These effects were observed even in absence of any E2 stimulation. Further we observed a significant overlap of differentially expressed genes in E2 stimulated and FBXO4 knockdown cells that were nullified by CCND1 siRNA. Conclusions: Our study elucidated that CCND1 is spared from degradation following loss of FBXO4 which in turn leads to a hyper-proliferative state. Our findings co-related with TCGA and METABRIC databases indicating a poor patient survival and relapse in FBXO4 low and CCND1 high cohorts of ER +ve breast cancer thereby implying a diagnostic value. Along with traditional hormonal therapy, an effective targeted therapy in breast cancer should be aimed to restore FBXO4 levels.
Utility of the Breast Cancer Index (BCI) in the clinical practice.
Authors: Kodali Saranya , Tipirneni Eswar , Dittus Kim
Published: May 2019

e14723

Background: Extended endocrine therapy (EET) greater than 5 years in early stage hormone receptor positive (HR+) breast cancer (BC) patients has shown benefit. However, EET is associated with side effects and there is no validated assay to determine which group of patients would derive benefit. Breast Cancer Index (BCI) is a validated bio-marker test that incorporates 2 distinct genomic assays and is prognostic/predictive. The objective of this study is to assess patient characteristics, pathologic features and patient preferences with regards to extending endocrine therapy after reviewing the BCI results. Methods: We performed a retrospective chart review on early stage HR+ BC patients from Jan, 2016 to Jan, 2017 at the University of Vermont Medical Center. We identified 25 cases on whom BCI was submitted. Results: Median age was 68 years. Majority of the patients were stage IA (64%). 56% of the tumors were moderately differentiated. All patients were ER +ve and 12% were HER2+. Median tumor size was 1.4 cm (0.3-4). 76% had poor tolerance to the ET and preferred the test to be sent. In LN-patients, BCI identified 42% as high risk and 52% as low risk for late recurrence and 32% who derive high benefit from EET. In LN+ patients, BCI identified 75% as high risk for late recurrence and 25% as low risk for late recurrence. 40% of the entire group were identified to highly benefit from EET (70% agreed to continue ET and 30% denied due to side effects). Conclusions: BCI is a reasonable test to consider in early stage HR+ BC, especially in patients with poor tolerance to ET. This test might aid in decision making with tolerability/compliance challenges to EET. [Table: see text]
HER2 targeted therapy and outcome in HER2-equivocal cases after 2018 ASCO/CAP HER2 guideline modification.
Authors: Crespo James , Sun Hongxia , Wu Jimin , Ding Qingqing , Tang Guilin , Robinson Melissa , Chen Hui , Sahin Aysegul A. , Lim Bora
Published: May 2019

e14729

Background: The best targeted therapeutic approach for HER2-equivocal cases remains unclear. New ASCO/CAP HER2 guidelines are intended to decrease this ambiguity by combining immunohistochemistry and in situ hybridization to resolve equivocal cases as positive or negative. However, the benefit of anti-HER2 therapy in HER2-equivocal cases is unknown. Methods: We retrospectively reviewed patients who visited MD Anderson from April 2017 to March 2018 with equivocal HER2 results based on the 2013 ASCO/CAP guidelines. The population was divided into 2 cohorts according to biopsy origin (primary cohort: biopsy from breast or axilla; recurrent/metastatic cohort: biopsy from recurrent or metastatic site). HER2 status was redefined using the 2018 ASCO/CAP guidelines. OS and PFS were calculated (Kaplan-Meier method) based on redefined HER2 status and use of HER2 targeted therapy. Results: A total of 139 equivocal results were found. Primary cohort had 90 patients (33 received neoadjuvant and 57 adjuvant therapy). HER2 IHC results were 0 (6.6%), 1+ (37.7%), 2+ (50%), 3+ (1.1%), and no IHC (4.4%). 94% of HER2-equivocal results became HER2 negative. Only 5 patients received anti-HER2 therapy, all of them in the HER2-negative group. After median follow-up of 1.91 yrs, 3 deaths and 8 progressions had occurred. There was no statistically significant association between anti-HER2 therapy and OS (p = 0.67) or PFS (p = 0.49). The recurrence/metastatic cohort had 49 cases with equivocal results. HER2 IHC results were 0 (6.1%), 1+ (22.4%), 2+ (26.5%), and no IHC (44.9%). 55% of HER2-equivocal results became HER2 negative, and only 1 patient received anti-HER2 therapy. After median follow-up of 2.96 yrs, 15 deaths and 35 progressions had occurred. There was no statistically significant association between anti-HER2 therapy and OS (p = 0.61) or PFS (p = 0.78). Conclusions: Most HER2-equivocal results were redefined as HER2 negative using the new ASCO/CAP guidelines. Association between anti-HER2 therapy and OS or PFS according to the new HER2 status was not observed. Although this is a small sample with short follow-up, patients with HER2-equivocal breast cancers seem to have clinical behavior similar to HER2-negative breast cancer.
ALK actionable mutations (muts) within cancer types and their responses to crizotinib (CZ).
Authors: Li Lin , Liu Caixia , Zhao Jun , Gen Lin , Dong Xiaorong , Yu Fenglei , Ai Xinghao , Huang Dongning , Chen Likun , Cheng Xiangyang , Kong Jinliang , Guo Renhua , Miao Liyun , Lin Li , Liu Li , Huang Gui , Wang Qiaoli , Chen Rongrong , Xia Xuefeng
Published: May 2019

e14739

Background: CZ was recommended by NCCN for ALK-rearranged metastasis NSCLC as first-line therapy. Knowledge on the occurrence of ALK actionable muts within cancer types and the responses of ALK fusion types to CZ could better guide the treatment of ALK-rearranged pts. Methods: 772 ALK-mutated pts (with any ALK muts) detected by tissue/ctDNA NGS in our institution from Oct. 2016 to Oct. 2018 were included. ALK+ (with any ALK actionable muts) and EGFR- (with no EGFR actionable muts) pts treated with CZ at any line were included for survival analysis. Progress-free survival (PFS) was estimated using Kaplan-Meier method and compared using log-rank test. Results: 339 ALK+ pts were analyzed, including 331 lung cancer (LC: 265 NSCLC; 1 SCLC; 65 Unknown histology), 3 brain cancer (BC), 1 colon cancer (CC), 3 cancer of unknown primary (CUP), and 1 with no clinical information. The most common ALK mut was fusion (331/339), including 315 EML4-ALK, 5 KIF5B-ALK, 4 STRN-ALK, 1 ACSL3-ALK, 1 CLIP4-ALK, 1 DCTN1-ALK, 1 HIP1-ALK, 1 SRBD1-ALK, 1 TFG-ALK, and 1 ZFP161-ALK. Fusion position was known for 308 EML4-ALK, including 132 V1, 108 V3, 32 V2, 19 V5’, and 17 other variants (Vx). Except for 2 V1 with CUP, 1 KIF5B-ALK with CUP, 1 Vx with CC, and 1 EML4-ALK (no variant information) with BC, all the other fusions occurred in LC. ALK F1174L was detected only in 2 pts, both of whom had BC, specifically neuroblastoma. CZ was given to 76 ALK+ EGFR- pts (mPFS: 9 mo, 95% CI: 8 – 13), including 70 EML4-ALK and 6 non- EML4-ALK (mPFS: 10 vs. 9 mo, p = 0.7). Though non-significant, V1 (n = 32) on average had a longer mPFS than V3 (n = 28) (12 vs. 8 mo, p = 0.3), which may partially be explained by the younger age at diagnosis of V1 than V3 (48 vs. 53, p = 0.005). 1 TFG-ALK, 1 CLIP4-ALK, 1 DCTN1-ALK, 1 HIPI-ALK, 1 ZFP161-ALK, and 1 KIF5B-ALK achieved a PFS of 3, 4, 8, 9, 13, and 19 mo on CZ, respectively. 1 SCLC (V3) received CZ at first-line progressed after 6 mo. Conclusions: Though majority of the ALK actionable muts were commonly found in LC, some alterations may occur more often in other cancer types, such as F1174L in neuroblastoma. Some fusion types (e.g. TFG-ALK) may benefit less from CZ than others (e.g. KIF5B-ALK). Limited by relative small sample size, these results merit further validation and investigation.
Tumor mutational burden reference materials for assay standardization.
Authors: Butler Matt , Konigshofer Yves , Clement Omoshile , Liu Li , Zhao Chen , Arcila Maria E. , Zehir Ahmet , Kohle Ravindra , Magliocco Anthony Martin , Gligorich Keith , Gloeckner Christian , Lovejoy Alexander F. , Hantash Feras , Sougnez Carrie , Lennon Niall , Anekella Bharathi , Garlick Russell
Published: May 2019

e14746

Background: Next Generation Sequencing based assays are designed to detect genomic aberrations in a limited number of target regions. However, there is a need for accurate measurement of tumor mutational burden (TMB) as low as 4 to as high as 50. As TMB assessment is added to various targeted panels, consistent results between panels are required to advance the broad use of this biomarker. Properly designed reference materials aid measurement standardization and are required to demonstrate assay concordance. We developed reference materials that vary in TMB score, tumor content 5-50% and are prepared in FFPE format. Methods: Seven lung and two breast tumor cell lines as well as matched “normal” lymphoblastoid cell lines were expanded in cell culture. Genomic DNA (gDNA) from each cell line was extracted. Tumor/normal mixes were made by mixing DNA and by embedding cells in FFPE blocks. Whole exome sequencing (WES) results were obtained using Agilent SureSelectXT for library construction and an Illumina Novaseq for sequencing. The Friends of Cancer Research TMB consensus method for analyzing WES data was used to filter variants and calculate TMB scores. Results: The cell lines were grown at large scale to produce extractable gDNA. 100% gDNA tumor, 30% gDNA tumor mixes and 30% FFPE cell line mixes were prepared. Preliminary results show that a clinically-relevant range of TMB values ranging from 4 to 35 mutations per million bases. The several thousand mutations that were observed across the lines were found in a variety of genes, which may explain why TMB in targeted panels is influenced by the specific target regions. Also, the initial results show that 30% cell line mixes showed similar TMB results to 100% gDNA. Conclusions: Our approach with wide ranging TMB values as tumor normal mixes is flexible and can be used to test different tumors and assays. For this study we established WES as the ground truth measurement for comparison to other assay formats and obtained comparison data from other panels. This approach also allows laboratories to test additional variables including formalin fixation, sample extraction, gene panel size, target regions, sequencing depth, filtering and limits of detection.
An aggressive RhoC phenotype is associated with relapse after external beam radiation therapy of a prostate cancer xenograft model.
Authors: Vilhelmsson Timmermand Oskar , Strand Sven-Erik , Ceberg Crister , Ceder Jens Adam
Published: May 2019

e14748

Background: Relapsed treatment resistant prostate cancer remains the second most common cause of male mortality in the western world. New therapies for therapy resistant prostate cancer depend on defining tumor cell sub-populations capable of surviving, reinitiating, and sustaining net growth of the cancer. In this study we focused on RhoC expression in a prostate cancer model after external beam radiation therapy (EBRT). RhoC has earlier been implicated in metastatic processes in most solid tumors. Methods: In mice, established s.c. LNCaP xenografts (n = 12) were irradiated, the beam collimated with lead to restrict it to the xenograft, with X-rays (200 kV, 1 Gymin-1, Gulmay Medical) for an absorbed dose of 15 Gy. Tumor tissue was collected after seven days or after 3-4 weeks when recurrent growth was seen. RhoC, AR, CD166 and Ki67 immunohistochemistry in tumor sections was visualized by using DAB or Peroxidase Green and nuclear red or haematoxylin blue as counter stain. Sections stained for RhoC and Ki67 were quantified with the Halo software (v2.3.2089.34) using the Indica Labs – cytonuclear v1.6 algorithm. The intensity of the RhoC staining was also quantified in a separate set of sections only stained for RhoC. Results: The RhoC staining intensity was significantly higher in relapsed xenografts (P = 0.003). Also, a significant difference in percentage of RhoC expressing cells in the two populations (P = 0.045) with 40.6 ± 5.8% (mean ± SEM) in relapsed tumors as compared to 15.0 ± 8.1% after 7d. The percentage of cells expressing Ki67 was not significantly different (P = 0.477). However, the percentage of double positive cells (Ki67, RhoC) was significantly higher in treated relapsed xenografts (P = 0.042). Further, a subpopulation of RhoC-positive cells were positive for AR and the stem cell marker CD166. Conclusions: Here we show that a subpopulation of proliferating RhoC-positive cells are more prevalent in relapsing PCa xenografts after EBRT, suggesting an aggressive phenotype that is associated with both proliferation and invasiveness. Another subpopulation of RhoC cells was found to express the prostate stem cell marker CD166, suggesting that pathways and properties involved in cellular self-renewal and invasiveness may be responsible for survival and recurrent growth in relapsed prostate tumors.