tbuckley/cabot-search huggingface dataset:
Inhibition of autophagy by MPT0L145, a novel PIK3C3 inhibitor, to overcome drug resistance in cancer cells.
Authors: Chun-Han Chen, Yu-Chen Lin, Yun Yen
Journal: Journal of Clinical Oncology (2018)

e14583 Background: PIK3C3 is a class III PI3K, which specifically phosphorylates phosphatidylinositol (PtdIns) at the 3 position of the inositol ring and generates phosphatidylinositol 3-phosphate (PtdIns3P). It in turn recruits WIPI proteins and is crucial to the initiation of autophagy, including nucleation and phagophore formation. Recently, several studies have shown that pro-survival autophagy is upregulated in cancer cells when exposing to chemotherapy and targeted therapy. MPT0L145, a highly potent PIK3C3 inhibitor, has been reported to impair autophagy in bladder cancer cells. Therefore, the rationale of this study is to combine MPT0L145 with agents that induce pro-survival autophagy to increase their therapeutic efficacy. Methods: The activity of PIK3C3 is inhibited by MPT0L145 (covered by PCT/US2016/043203) or stably knocking down by shRNA in cancer cell lines and combined with clinical therapeutic agents reportedly induce pro-survival autophagy. Cell viability was examined by MTT assay and combination index was determined by Compusyn software. The autophagy flux was evaluated by detecting protein levels of LC-3II and p62 through western blot analysis. Results: The ability to induce autophagy by gefitinib and gemcitabine was confirmed as evidence by the increase of LC3-II and decrease of autophagy substrate, p62. Both of PIK3C3 knockdown and administration of MPT0L145 synergistically sensitize the killing effects of gefitinib and gemcitabine in A549 cells and Panc1 cells, respectively. However, the protein levels of LC3-II and p62 were increased, suggesting an incomplete autophagy where the function of PIK3C3 was impaired. Moreover, no appreciable apoptotic cell death was observed. Conclusions: The combination of MPT0L145 and autophagy inducers showed synergistic effects in vitro. This study revealed a potential strategy by targeting PIK3C3 to overcome drug resistance associated with pro-survival autophagy in cancer therapy.
Sarcopenia and toxicity of the anti-PD1 inhibitors in real-life lung cancer patients: Results from the French Nationwide SCAN study.
Authors: Marie-Pierre Revel, Bruno Raynard, Frederic Pigneur, Mario Di Palma, Alain Toledano, Elise Deluche, Olivier Romano, Francois Goldwasser
Journal: Journal of Clinical Oncology (2018)

e21066 Background: The SCAN study is a French nationwide study which aimed to measure the prevalence of sarcopenia in real-life cancer patients (pts). A secondary objective was to study the relationship between sarcopenia and the occurrence of treatments toxicities. The subgroups of lung cancer pts and pts under anti-PD1 are shown. Methods: This prospective study lasted over 2 months from Sept 2017, included pts of 29 private and public hospitals with either lung, prostate, breast, colon, or renal cancer, an ongoing treatment, and an available CT-scan to assess muscle mass. Total muscle area (TMA), skeletal muscle index (SMI) were measured from axial L3 CT sections. Clinical characteristics included PS status, dietary intakes, measurement of muscle strength using a JAMAR hydraulic hand dynamometer. Any grade of toxicity leading to treatment interruption was considered as acute limiting toxicity (ALT). Results: Of the 779 (25.8%) pts included in SCAN, 201 had metastatic lung cancer, 195 were assessable for toxicity, 198 for muscle mass. Median age was 65 years (19-87), and there were 120 males (60%). Half of pts (53%) were treated with cytotoxic chemotherapy, 38% with immunotherapy and 9% with targeted therapy. The PS status was 0-1 in 70% of pts. The prevalence of sarcopenia was 70%. Severe anorexia (dietary intakes≤4/10) was more frequent under chemotherapy than under anti-PD-1: 21.2% vs 1.4% (p < 0.01). Few pts experienced ALT: 10/195 (5.1%). Among them, 80.0% had lost weight within the last 30 days. In this group, the occurrence of ALT was 11.4% (8/70) versus 1.7% (2/118) in the pts without weight loss (p = 0.01). Considering all treatments, there was a reproducible, non-significant, association between muscle loss, using TMA or SMI, reduced muscle strength, and occurrence of ALT. In the 115 pts under anti-PD1 (77 lung, 38 renal cancer), the ALT were 5/40 (12.5%) in pts with SMI ≤41 cm²/m² vs 1/68 (1.5%) in pts with SMI ≥42 cm²/m² (p = 0.05). The odds ratio was 7.74 (95%CI: [1.01-92.79]; p = 0.024) in pts with TMA < 144cm². Conclusions: Sarcopenia is frequent in lung cancer patients. Sarcopenia is associated with higher risk of immunotherapy interruption.
PD-L1 assessment in FNA (EBUS) dervied samples.
Authors: Andrew Lerner, Lonny Yarmus, David Feller-Kopman, Hans Lee, Jesi Oswiecimka, Patrick M. Forde, Peter B. Illei
Journal: Journal of Clinical Oncology (2017)

11615 Background: Pembrolizumab therapy for non-small cell lung cancer requires PD-L1 immunohistochemistry (IHC). FDA approval was based on staining of resections or core biopsies. Little is known about PD-L1 expression in fine needle aspiration (FNA) specimens including EBUS of mediastinal lymph nodes. Methods: IHC was performed using the PD-L1 IHC 22C3 pharmDx test on formalin fixed paraffin embedded FNA cell blocks of 8 squamous cell carcinomas and 15 adenocarcinomas (ACA) (age: 42-84, mean: 63, median: 63; 11 female and 12 male). Mutation data (50 gene NGS panel, and ALK, ROS1, RET and MET FISH) was available for 14 ACA. Membranous PD-L1 staining of any intensity and extent was recorded in at least 100 tumor cells (tumor proportion score). The tumors were grouped as: no staining (&lt;1%), low expression (1-49%) and high expression (50% or more). Results: Six (26%) tumors showed no staining, 6 (26%) low expression and 11 (48%) high expression (table 1). High PD-L1 expression was seen in 1 ALK, 1 braf, 1 EGFR, 1 Her2 exon20, 2 kras mutated tumors and 2 tumors with no or unknown mutation status, low PD-L1 expression in 1 braf and 1 EGFR mutated case and 1 tumor without mutation. No expression was seen in 1 ROS1, 1 kras mutated tumor and 1 tumor without mutation. One EBUS biopsy also had PD-L1 assessed on a transbronchial biopsy of the primary tumor showing similar staining (30% versus 20%). Conclusions: PD-L1 immunohistochemistry appears to be feasible using cell blocks of fine needle aspirates containing adeqaute number of viable tumor cells. The majority of ACA were also adequate for NGS mutation and FISH analysis. High PD-L1 expression was seen in approximately half the tumors, which is greater than has been observed in resections/core biopsies, this finding merits further study. High expression of PD-L1 was seen in both squamous cell carcinoma and adenocarcinoma, and tumors with and without common driver mutations. [Table: see text]
Abstract 11905: Loss of Adrenergic and Adenosine Regulation of Extradyadic L-type Calcium Channels in Rat Atrial Myocytes in Heart Failure
Authors: Alexey V Glukhov, Marina Balycheva, Jose Sanchez-Alonso, Navneet Bhogal, Ivan Diakonov, Marta Mazzola, Giuseppe Faggian, Julia Gorelik
Journal: Circulation (2015)

It has recently become evident that discrete clusters of L-type Ca2+ channels (LTCCs) exist along the cardiomyocyte sarcolemma in association with distinct membrane structures. Such microdomain-specific localization impact channel function and regulation by a variety of neurohormonal pathways, including adrenergic and adenosine. Disruption in their subcellular targeting may contribute to the pathophysiology of cardiac diseases, including heart failure (HF). We used Ca2+ imaging and super-resolution scanning patch clamp to examine microdomain-specific regulation of LTCCs in atrial myocytes isolated from control and 16-weeks post-MI HF rats. In control, β1 adrenergic receptors (β1ARs) stimulation (ISO 100 nM and β2AR antagonist ICI 50 nM) enhanced spontaneous Ca2+ release events while the following adenosine A1 receptors (AdoA1Rs) stimulation (2'-MeCCPA 200 nM) abolished β1ARs effects. Non-localized β1ARs stimulation (ISO 2 μM and ICI 50 nM in external solution) activated single LTCCs in both t-tubules and on the crest of sarcolemma, increasing LTCC occurrence and enhancing their open probability and amplitude. Local AdoA1R stimulation through the pipette (10 μM) on β1ARs-pretreated cells completely abolished β1ARs effects. In HF, both β1ARs and AdoA1R effects on LTCCs were preserved in t-tubules and lost on the crest. Similar, the effect of β1AR stimulation on spontaneous Ca2+ release events was significantly reduced in HF (28% vs 134% in HF vs control, P&lt;0.01) and the anti-adrenergic effect of AdoA1R was lost (8% vs 84% in HF vs control, P&lt;0.01), despite up-regulation of β1 and AdoA1Rs mRNA. No changes in caveolae density were observed in HF (4.6±0.4 caveolae/μm vs 3.7±0.5 caveolae/μm for HF vs control, P=0.167). Our findings provide the first direct evidence for microdomain-specific remodeling of LTCC regulation by adrenergic and adenosine receptors in HF which could be linked to local disruption in the interactions between LTCCs and their cellular microenvironment. This extends beyond the classical concept of electrical remodeling, stressing the involvement of alteration of spatial compartmentation of ion channels and receptors in addition to classical concepts of changes in protein expression and post-translational modifications.
Cisplatin to induce cancer stem cell state in ovarian cancer.
Authors: Caner Saygin, Andrew Wiechert, Praveena Thiagarajan, Vinay Rao, James Hale, Masahiro Hitomi, Analisa DiFeo, Justin Lathia, Ofer Reizes
Journal: Journal of Clinical Oncology (2016)

e17098 Background: Ovarian cancer (OC) is the most lethal gynecological cancer. Despite high rates of response to initial cytoreductive surgery and taxane-platinum combination therapy, vast majority of patients recur. OC is marked by a high degree of cellular heterogeneity and contains a cancer stem cell (CSC) population that contributes to tumor growth, recurrence, and treatment resistance. Methods: We used isogenic OC cell lines A2780/CP70 (cisplatin naïve/resistant). A lentiviral construct, which contained GFP attached to the promoter region of the CSC transcription factor NANOG, was used to transduce them. Stably transduced cells were sorted based on their GFP intensity. Tumorsphere formation assays and tumor initiation experiments were performed to assess self-renewal. Sorted cells were cultured, treated with cisplatin, and counted for 5 days. Time-lapse imaging was performed to monitor GFP expression. Results: A2780 GFP+ cells expressed higher levels of CSC markers including CD44 and CD133. These cells had higher levels of NANOG and OCT4. Moreover, CP70 cells displayed higher NANOG and OCT4 expression as compared with A2780 cells. A2780 GFP+ cells were significantly more self-renewing than their GFP- counterparts. In addition, CP70 cells possessed higher self-renewal as compared to A2780. The gold standard functional CSC assay is tumor initiation and upon sorting and injection into NSG mice, all mice injected with GFP+ cells developed tumors whereas in mice injected with 50K and 5K GFP- cells, 4/5 and 3/5 developed tumors, respectively (CSC frequencies were 1:1; CI = 1:6,271-1:1, and 1:17,979; CI = 1:49,395-1:6,544 in GFP+ vs GFP- cells, respectively). Moreover, we compared cisplatin sensitivity between cultured GFP+ and GFP- A2780 cells, and found a significant survival advantage in GFP+ over GFP- cells. Upon cisplatin treatment, surviving GFP- cells showed higher GFP intensity and NANOG mRNA expression as compared to untreated cells. This was confirmed by visual induction of GFP using time-lapse microscopy. Conclusions: Cisplatin may serve as an inductive pressure for the stem cell state and enrich CSCs. Applying this knowledge to current OC treatment paradigms, partnering cytotoxic chemotherapy with a CSC targeted therapy, is paramount.
NIMG-04. DIFFUSION RESTRICTION ON MR IMAGING IN THE T2 HYPERINTENSE, BUT OTHERWISE NORMAL-APPEARING WHITE MATTER OF GLIOBLASTOMA PATIENTS TREATED WITH TTFIELDS CORRELATES WITH SURVIVAL
Authors: Josef Vymazal, Radovan Zacek, Jan Sroubek, Aaron M Rulseh
Journal: Neuro-Oncology (2018)

Based on the positive results of our pilot study that evaluated MR diffusion tensor imaging characteristics in the white matter of 13 subjects with glioblastoma, we aimed to evaluate the prognostic value of diffusion restriction in a new, larger cohort of glioblastoma patients. Thirty-five subjects diagnosed with glioblastoma were treated by surgical resection, radiotherapy, temozolomide and TTFields (Optune®). All subjects were followed by MRI at 1.5 T approximately every two months (total range 160–1810 days) with standard imaging, including diffusion-weighted imaging, until tumor progression. Apparent diffusion coefficient (ADC) maps were calculated using FSL and freehand ROIs were placed on B0 images within the T2 hyperintense white matter (T2HWM) near the resection cavity by a single investigor (AMR), and then projected onto the ADC maps. Additionally, an ROI was similarly placed within the normal-appearing white matter (NAWM). Final values (ADC-ratio) were calculated by dividing ADC in T2HWM by ADC in NAWM. Linear regression slopes (ADC-ratio vs. time) for individual subjects were fitted in R. In 26 subjects a positive slope was observed in ADC-ratio over time, while in the remaining subjects the slope was negative. We found a significant difference between subjects with positive and negative regression slopes in overall survival (OS; hazard ratio = 0.27 [0.10.69], log rank p = 0.009) and progression-free survival (PFS; hazard ratio = 0.31 [0.120.81], log rank p = 0.02), with positive regression slope associated with more favorable outcome. Our data suggest that increasing ADC in the affected (T2 hyperintense) white matter of glioblastoma patients is favorable, with significantly greater PFS and OS in patients with increasing ADC over time. Changes in ACD likely reflect changes in tumor density in the affected white matter, with decreasing values (increasing diffusion restriction) corresponding to tumor progression.
Single-center "real life experience" with pembrolizumab (PEMBRO) in pretreated advanced melanoma patients.
Authors: Yanina Jansen, Max Schreuer, Bart Neyns
Journal: Journal of Clinical Oncology (2016)

e21049 Background: PEMBRO, a PD-1 blocking mAb, is approved for the treatment of patients (pts) with advanced melanoma. Outcome of pts treated outside a clinical trial is not established. Detection of cancer mutations on circulating cell-free DNA (cfDNA) holds promise as a prognostic biomarker. Methods: prospective academic study on the outcome of pretreated advanced melanoma pts in a PEMBRO expanded access program (EAP). BRAF V600 mutations in cfDNA were assessed by IdyllaTM ctBRAF Mutation Prototype Assay (Biocartis). Results: between Sep 2014 and Jan 2016, 111 pts initiated PEMBRO (2 mg/kg q3wks). Baseline characteristics: 42M/69F; median age 58y (range 26-93); 86x primary skin, 4x mucosal, 14x unknown and 7x uveal; WHO-PS 0/1/2: 72-, 27-, 12 pts; AJCC Stage: 9x IIIC, 5x IV-M1a, 8x -M1b, 89x M1c; 55 pts BRAF V600mut; 34 pts with brain metastases; LDH > ULN: 48 pts; CRP > ULN: 55 pts; ALC < 1200/mm³: 32 pts. In addition to IPI, most freq. prior therapies: BRAF(+MEK) inhib. (47 pts), chemotherapy (30 pts), and RT for brain metastases (20 pts). Median follow-up: 24 wks and PEMBRO treatment was ongoing in 53 pts. A PEMBRO related AE of any grade occurred in 75 pts (68%) and a gr3/4 PEMBRO related AE in 12 pts (11%) (most freq.: 3x colitis, 3x hepatitis, 3x fatigue). All PEMBRO-related AEs gr > 3 were reversible upon PEMBRO interruption, corticotherapy, and infliximab (1pt). There was no gr5 AE. Among the 74 pts (excl. uveal) evaluable for response, ORR was 23% (8 CR, 9 PR; incl. 2 pts with brain metastases). Median PFS (excl. uveal): 12 wks (95% CI 7-16); 24- and 52 wks PFS%: 39 and 35% respectively; median OS has not been reached. In multivariate analysis, brain metastases and BRAF V600mut were correlated with worse PFS; brain metastases and CRP > 5xULN with OS (Cox logistic-regression, P < .05). Idylla Cq values on 72 plasma samples correlated well (R² = 0.86, p < 0.05) with BRAF V600E droplet counts on ddPCR (Bio-Rad). No correlation was found between baseline positive BRAF V600mut ctDNA (N = 13; 42%) and survival in 31 assessable pts with BRAF V600mut melanoma. Conclusions: this single-center "real life" experience with PEMBRO in an EAP for pretreated advanced melanoma pts strongly resembles the profile of PEMBRO observed in prospective trials.
Targeted tumor profiling and actionable somatic variants in sarcoma.
Authors: Eytan Ben Ami, Matthew Louis Hemming, Michael Nathenson, Katherine Anne Thornton, Priscilla Merriam, Jeffrey A. Morgan, Chandrajit P. Raut, Suzanne George, Andrew J. Wagner, George D. Demetri
Journal: Journal of Clinical Oncology (2018)

11551 Background: The impact of next generation sequencing data on treatment decision and clinical outcome in sarcoma remains under investigation. Methods: We queried the Dana Farber Cancer Institute database to identify soft-tissue and bone-sarcoma patients (pts) who underwent targeted sequencing under a research protocol. We searched for mutations and somatic copy number alterations (SCNA) and evaluated clinical outcomes in cases bearing actionable genetic abnormalities (eligible for NCI-MATCH/ASCO-TAPUR studies). Results: 613 pts with 38 sarcoma histologies were evaluated. Leiomyosarcoma (LMS, 23.8%), liposarcoma (LPS, 13.2%), undifferentiated pleomorphic sarcoma (9.4%) and solitary fibrous tumor (4.7%) were the common histologies. Frequent mutations were observed in TP53 (30.1%), ATRX (12.1%) KMT2D (11.1%), NF1 (8%) and ATM (7.3%) genes. Common SCNA included homozygous deletions in RB1 (10%) and CDKN2A (8.1%), and high copy number gains in MDM2 (13.6%), CDK4 (11.5%), GLI1 and MYC (3%). 123 pts (20%) had 145 actionable somatic variants (59 mutations, 86 SCNA). Actionable mutations, observed in 52 pts, frequently involved the PI3K/Akt/mTOR pathway (PI3K3CA, TSC1/2 and AKT1 genes; 25% of actionable mutations), BRCA1/2 (15.2%), and IDH1/2 (17%, exclusively in chondrosarcoma) genes. Actionable SCNA, observed in 82 pts, comprised mostly of homozygous deletions in CDKN2A (51% of actionable SCNA) and PTEN (15.1%), and high copy number gain, not associated with LPS, in CDK4 (12.8%). Of 72 metastatic pts with actionable alterations and clinical data, 12.5% (9 pts) received matched therapies. Tumor responses were observed in 6 pts, including durable responses (6-28 months) in dedifferentiated chondrosarcoma (IDH2 mutation), inflammatory myofibroblastic tumor (ALK rearrangement), LMS (TSC2 mutation), spindle cell sarcoma (NTRK1 rearrangement), and chordoma (CDKN2A deletion). Conclusions: As many as 20% of sarcoma pts may harbor actionable genetic alterations. We observed durable tumor responses with matched therapies in several sarcoma histologies and further research is needed to understand the proportion of patients who may benefit from this approach. Mutations and SCNA incidence by histology will be presented at the meeting.
Trends in non-small cell lung cancer biomarker testing rates in the US and Western Europe from 2014-2017.
Authors: Bijal Shah-Manek, Chitra Karki, Sarah Whitmire, Long Ha, Samantha Martino, Valerie Wriede, Ayse Levent
Journal: Journal of Clinical Oncology (2018)

e20515 Background: Molecular diagnostics have been developed and used as potential predictors of responsiveness to personalized targeted therapies, but research is still ongoing. Using data from Ipsos’ Global Oncology Monitor, this study characterizes the trends of diagnostic testing in both the US and Europe over 3 years. Methods: A multi-country retrospective chart review of non-small cell lung cancer (NSCLC) patients in the US and 5 European (EU5) countries (France, Germany, Italy, Spain, UK) was conducted between 7/2014-6/2017. Year 1 was defined as 7/2014-6/2015, and years 2 and 3 were defined similarly in the corresponding years. Physicians randomly selected patient charts currently on an anti-cancer regimen and extracted data on patient demographics, disease and biomarker status. Testing rates of EGFR mutation (EGFRm), ALK, PD-L1, ROS-1, and KRAS were analyzed. Data for year 1 was not available for PD-L1 and ROS-1. Sample size varied by test and year. Chi-square test for trend was used to analyze biomarker testing rates. Results: The rates of all 5 biomarker tests increased in both the US and EU5 samples (p < 0.0001). In the US cohort, PD-L1 testing experienced the highest growth from years 2 to 3, with an increase from 19% (n = 857/4508) to 46% (n = 1861/4046). KRAS testing experienced the highest growth in the EU5 cohort, with an increase from 38% (n = 2809/4582) to 59% (8072/5610) over 3 years. The absolute growth rates of ROS-1 were similar between the US (11%) and EU5 (13%) cohort. EGFRm and ALK testing rates were high with slower growths in both cohorts. ALK testing experienced the slowest growth in the US cohort, with an increase from 63% (n = 2639/4189) to 70% (n = 2832/4045), and EGFRm testing experienced the slowest growth in the EU5 cohort, with an increase from 71% (n = 5263/7413) to 81% (n = 11086/13687) over 3 years. Conclusions: Diagnostic testing is increasingly becoming commonplace in the treatment of NSCLC. In both the US and EU5 cohorts, the majority of patients received EGFRm and ALK testing; and testing rates continue to increase in both groups. Although PD-L1 and ROS-1 testing rates were far less common, they experienced the greatest growth in years 2 and 3 of the study period.
Alterations in the uterine microbiome in patients with early endometrial cancer: Variations by ethnicity and obesity.
Authors: Leslie Horn Clark, Temitope O Keku, Nicole Amber McCoy, Gabrielle Hawkins, Victoria Lin Bae-Jump, Wendy R. Brewster
Journal: Journal of Clinical Oncology (2017)

e17114 Background: Obesity and race are known to negatively impact clinical outcomes in endometrial cancer (EC). Mounting evidence suggests the gut microbiome may play an important role in the pathogenesis of both obesity and cancer, however, little is known about the microbiome of the malignant uterus. Further, there is known to be microbiome variability by ethnicity in the vagina and gut. Thus, we sought to evaluate the microbiota of ECs and to assess for variations by obesity and race. Methods: Following IRB approval, banked tumor specimens of patients undergoing hysterectomy for management of endometrioid EC were identified. Tumors were analyzed from African-American (AA) and Caucasian (CAU) patients and stratified as obese (BMI ≥ 30kg/m 2 ) or non-obese (BMI &lt; 30kg/m 2 ). The microbiota of the endometrial tumors were characterized by 16S rRNA high throughput sequencing. Descriptive statistics were performed. Results: We evaluated specimens from 11 AA (52%) and 10 CAU (48%). The median age was 68.9 years. Majority of women were postmenopausal (n=17, 81%) and obese (n=14, 66.7%). Tumor grade was evenly distributed with 7 grade 1 (33.3%), 6 grade 2 (28.6%), and 8 grade 3 tumors (38.1%). We found that EC microbiota richness was higher in AA than CAU (p=0.02). When analyzed by race and obesity status, we observed significant differences in EC microbiota composition. The microbiota composition differed between obese AA and obese CAU (p=0.04). In obese subjects the distribution of Proteobacteria was 55% AA vs. 30.7% CAU (p=0.02) while Actinobacteria abundance was 17.8% versus 34.4% (p=0.05) (Table 1). Among CAU, the EC microbiota profile differed between obese and non-obese (p=0.07). At the genus level, the relative abundances of Delftia, Finelgoldia, Geobacillus, Propionibacterium and Pseudomonasvaried by obesity and race. Conclusions: Bacteria exist in the malignant uterus. EC microbiota diversity was greater in tumors of AA versus CAU women, and tumor microbiota profiles were distinct between ECs of obese and lean women. Better understanding of the inter-relationship of obesity and race on the EC microbiota may provide critical insight into the disparate clinical outcomes between AA and CAU women with EC.