bluuebunny/crossref_metadata_2025 huggingface dataset:
A landscape analysis of immunotherapy-related biomarkers across solid tumors.
Authors: Kujtan Lara Ann , Morris Scott , Masood Ashiq , Subramanian Janakiraman
Published: May 2019

e14268

Background: Checkpoint inhibitor-based immunotherapy has varying success among tumor types and patients. Predictive biomarkers are in high demand to assist with patient selection. Responses to these agents are correlated with programmed cell death ligand 1 (PD-L1) expression, microsatellite instability (MSI), and tumor mutational burden (TMB). Here we evaluated PD-L1 expression, MSI status and TMB in a variety of solid tumors to determine their relationships. Methods: A total of 109 specimens were identified in patients diagnosed with solid tumors that underwent a Paradigm Diagnostic Cancer Test. MSI scores were determined by the number of indels present in runs of 6 homopolymer bases per megabase, with a cut-off of 6 to distinguish between MSI-high (MSI-H) and MSI-stable (MSI-S). TMB scores greater than or equal to 10 muts/Mb were designated as high (TMB-H). Results: Of all tumors, 19.3% were MSI-high. PD-L1 testing was performed in 71.5% of all samples; of these, 21.8% were PD-L1 positive. TMB scores of PD-L1 positive tumors (mean = 10.3 muts/Mb) and PD-L1 negative tumors (mean = 8.6 muts/Mb) did not differ significantly (p = 0.57). All MSI-H tumors were TMB-H, and MSI-H tumors had higher mean TMB scores than MSI-S tumors (57 muts/Mb versus 7.6 muts/Mb; p = 0.06). Among TMB-H tumors, MSI-H status was associated with higher TMB scores compared to MSI-S tumors (55.2 muts/Mb versus 22.1 muts/Mb, p = 0.18). Of the both MSI-S and TMB-H tumors, 68.8% were lung cancers and 18.8% were breast cancers. Tumors both MSI-H and TMB-H were 47.6% gastrointestinal carcinomas and 33.3% endometrial carcinomas. Conclusions: In our analysis, TMB was independent of PD-L1 status. All MSI-H tumors were also TMB-H; these were primarily gastrointestinal and endometrial carcinomas whereas TMB-H tumors that were MSI-S consisted predominantly of lung cancers. This difference may be due to the different mechanisms of acquiring mutations during carcinogenesis in these two tumor types. These data also highlight the need for integrated PD-L1, MSI and TMB testing to identify potential responders to immunotherapy.
Preliminary effects on the innate immune system with the combination of cetuximab and durvalumab in metastatic/ relapsed head and neck squamous cell carcinoma in a phase II trial.
Authors: Gulati Shuchi , Vachon Rachel , Desai Shireen , Steele Aubrey , Palackdharry Sarah , Takiar Vinita , Wise-Draper Trisha Michel
Published: May 2019

e14273

Background: Cetuximab is a recombinant chimeric monoclonal IgG1 antibody which binds specifically to the epidermal growth factor receptor (EGFR) and stimulates an innate immune response by promoting natural killer (NK) cell mediated antibody-dependent cell-mediated cytotoxicity (ADCC). Cetuximab is approved as a single agent in relapsed/metastatic head and neck squamous cell carcinoma (R/M HNSCC). PD-1 check-point inhibitors which release the inhibition of the adaptive immune response, are also approved as single agents in this setting. However the response rates with these drugs, when used individually range from 10-20%. We hypothesized that adding a PD-L1 inhibitor, durvalumab to cetuximab would cause anti-tumor synergy by activating the innate as well as adaptive immune systems without compromising safety in this phase-2 trial in R/M HNSCC patients who have progressed on platinum based therapy. Methods: Blood samples were collected from the first six enrolled patients prior to starting treatment and 4 weeks after the first combined dose of cetuximab and durvalumab. PBMCs were isolated, stained with a live/dead stain as well as CD3, CD4, CD8, CD56, CD16 and NK2GD (natural killer group 2 member D activation receptor) antibodies and analyzed by flow cytometry. Cytokine levels in plasma were measured using standardized ELISA assay kits. Results: Compared to pre-treatment levels, post-treatment samples showed an increase in activated cytokine producing NK cells (CD56bright/CD16-) in all but one patient. Activated cytotoxic NK cell subpopulations (CD56dim/CD16+) showed variable results post-treatment. CD8+ T cells were similar pre and post-treatment in 5 patients. TGF-b levels increased in 5 patients and decreased in 1 patient post-treatment. Interestingly, the patient with decreased TGF-b levels post-treatment had an almost doubling of CD8+ T-cells and an increase in activated cytokine producing NK cells (CD56bright/CD16-). Conclusions: The clinical trial is ongoing and therefore, comparison to clinical response has not yet been analyzed. However, these findings support the combination of cetuximab and durvalumab in R/M HNSCC given the activation of an NK-cell mediated innate immune response in these patient samples. Clinical trial information: NCT03691714.
Comprehensive analysis of immune indicators provides prognostic markers for early stage non-small cell lung cancer.
Authors: Ding Xiao , Cui Jiuwei
Published: May 2019

e14276

Background: Adjuvant chemotherapy has been established as a standard for patients with early stage NSCLC after complete tumor resection, but the 5-year survival rate can only increase by about 5%. Given its high morbidity and mortality, new biomarkers are needed to predict clinical response and prognosis of resected NSCLC. The expression of MHC-I, PD-L1 and CD8+ TILs were detected in this study to investigate the potential prognostic markers in patients with early stage NSCLC. Methods: Tissue of 125 patients with surgically resected NSCLC (stage I-III) were obtained from the First Hospital of Jilin Universtiy. MHC-I, PD-L1 and CD8+ TILs were detected with immunohistochemistry. The association between their expression levels and patients’ prognosis was analyzed. Results: MHC-I was down-expressed, and PD-L1 was up-expressed in NSCLC tissues compared with the para-cancer tissues. CD8+ TILs could be seen in tumor stroma and nest. PFS and OS was poorer in MHC-Ilow group compared with MHC-Ihigh group (P < 0.05). PFS and OS in PD-L1− group tended to be longer than the PD-L1+ group, but the difference was not significant (P > 0.05). For CD8+ TILs, PFS and OS of the CD8inflamed group were longer than the CD8non-inflamed group (P < 0.05). With multivariate analysis, we found that MHC-I and CD8+ TILs might be independent prognostic factors of NSCLC (P < 0.05). Based on the PD-L1 and CD8+ TILs expression, we divided the patients into four subgroups: PD-L1+/CD8inflamed, PD-L1+/CD8non-inflamed, PD-L1−/CD8inflamed, and PD-L1−/CD8non-inflamed. Statistical differences were achieved both in PFS and OS (P < 0.05). PFS and OS of the PD-L1+/CD8inflamed and PD-L1−/CD8inflamed group were longer than the other two groups. Patients with PD-L1+/CD8non-inflamed experienced the worst PFS and OS. Besides, we divided the patients into four subgroups according to their MHC-I and CD8+ TILs expression conditions: MHC-Ihigh/CD8inflamed, MHC-Ihigh/CD8non-inflamed, MHC-Ilow /CD8inflamed, and MHC-Ilow/CD8non-inflamed. Statistical differences were achieved both in PFS and OS of these subgroups (P < 0.05). PFS and OS of the MHC-Ihigh/CD8inflamed group were longer than the other three groups. Patients in MHC-Ilow/CD8non-inflamed group experienced the worst PFS and OS. Conclusions: It was reported for the first time that the combination of PD-L1 and CD8+ TILs, MHC-I and CD8+ TILs suggested impressive prognostic values in early stage NSCLC in this study. The comprehensive analysis of immune indicators may provide prognostic markers for early stage NSCLC.
Prognostic value of Tfh-like cells in breast cancer.
Authors: Ma Linxiaoxi , Guo Liang , Goswami Shyamal , Zhang Xiaoming , Wu Jiong
Published: May 2019

e14285

Background: The treatment of breast cancer, one of the most common malignant tumors for female, focuses on the combined treatments based on subtypes including Luminal A, Luminal B, HER2 overexpression and triple negative. Given that none of those subtypes takes into consideration of immunological parameters, it is valuable to explore the biological characteristics and prognostic value of immune cells in breast caner. T follicular helper cells (Tfh cells) have been identified in different types of tumors with rare datas in breast cancer. One of them indicated the positive prognostic value of 8-gene Tfh signature. Exploration of Tfh cells may provide new clues to the potential immunotherapies in breast cancer. Methods: Freshly resected invasive breast cancer tissue from Fudan University Shanghai Cancer Center were collected during 06/2015 to 05/2017. Patients were divided into Luminal A (N = 61), Luminal B (HER2-) (N = 138), Luminal B (HER2+) (N = 72), HER2+ (non-luminal) (N = 57) and triple negative (N = 58) based on St Gallen International Expert Consensus. Results: A subpopulation of PD1hiCD4+ T cells in the tumor tissues of breast cancer was identified as Tfh-like cells, with the specific expression of Bcl-6 and CXCL13, confirmed by IHC. Phenotypes of those cells were checked by flow cytometry. Tfh-like cells were of a high level of ICOS but negative for CXCR5, suggesting that they were atypical Tfh cells. Data showed that high grade (N = 166; mean±SEM, 17.62±0.87) compared with low grade (N = 220; mean±SEM, 12.41±0.46) or high Ki-67 level (N = 300; mean±SEM, 15.38±0.58) with low Ki-67 level (N = 86; mean±SEM, 12.10±0.67), and negative ER expression (N = 115; mean±SEM, 17.47±1.05) compared with positive ER expression (N = 271; mean±SEM, 13.46±0.50) or negative PR expression (N = 177; mean±SEM, 16.87±0.83) compared with positive PR expression (N = 209; mean±SEM, 12.77±0.50) was associated with higher frequency of Tfh-like cells (P < 0.01 to all). Patients with triple negative had higher frequency of Tfh-like cells than those with Luminal A (difference, 6.83; P = 0.0006) and Luminal B (HER2-) (difference, 4.61; P = 0.0124). Patients with HER2+ (non-luminal) had higher frequency of Tfh-like cells than those with Luminal A (difference, 5.34; P = 0.0146). These results indicate that higher frequency of Tfh-like cells could have negative prognostic influence. Conclusions: Our study defined a PD1hiBcl6+CXCL13+CD4+ T cell subpopulation, which specifically secretes cytokine IL-21 as Tfh-like cells. Higher frequency of Tfh-like cells is more likely to be seen in patients with poor prognosis.
Prediction the clinical outcomes of cancer patients after peptide vaccination.
Authors: Toke Eniko Rita , Megyesi Mónika , Molnar Levente , Tóth József , Lőrincz Orsolya , van der Burg Sjoerd H. , Welters Marij , Melief Cornelis Joseph , Schönharting Wolfgang , Urban Sybille , Roehnisch Tim , Heine Uwe , Somogyi Eszter , Csiszovszki Zsolt , Pántya Katalin , Páles Péter , Miklós István , Lori Franco , Lisziewicz Julianna
Published: May 2019

e14295

Background: Neoantigen vaccines can activate T-cells that specifically kill tumor cells. However, most vaccine peptides, selected either as HLA-binders or as HLA-presented epitopes, do not induce T-cell responses in HLA allele-matched individuals. We hypothesized that personal-epitopes (PEPIs) binding to multiple autologous HLA-alleles induce potent T-cell responses. Methods: Epitopes binding to single HLA and PEPIs binding to 3 autologous HLA-alleles were identified with our novel and validated PEPI Test (CE Marked) and correlated with CD8+ and CD4+ T-cell responses of (pre)malignant patients vaccinated with Synthetic Long Peptides (SLPs) in 2 clinical trials. A Model Population of 433 HLA-genotyped individuals was used to determine the percentage of subjects with PEPIs from SLPs (PEPI-Score). As a proof of principle, a personal vaccine was developed matching with 14 HLA-alleles of a cancer patient with PEPIs from 12 tumor-antigens. T-cell reactivities were tested by interferon-γ ELISPOT. Results: There was no correlation between single HLA-binding epitopes and HPV-specific T-cell responses of patients. In contrast, there was 90% and 69% agreement between HLA-class-I PEPIs and CD8+ T-cell responses (p < 0.001) and HLA-class-II PEPIs and CD4+ T-cell responses (p = 0.005), respectively. PEPI-Score predicted the T-cell response rate of SLP vaccine clinical trials. As predicted by the PEPI Test, personal vaccine treatment activated tumor-specific T-cells: 91% and 100% of the vaccine peptides elicited CD8+ and CD4+ T-cell responses, respectively. Conclusions: A patient’s HLA genotype is a major determinant of vaccine responses and PEPIs are genetic biomarkers of T-cell responses. PEPI Test prediction of vaccine responses can be utilized by clinicians for selecting the vaccine treatment and for the development of highly immunogenic personal vaccines matched to a patient’s complete HLA genotype (NCT03391232).
Analysis of Chinese acral and mucosal melanoma patient genomic and neoantigen profiles in cancer vaccine development: A pilot study.
Authors: Guo Xianling , Gao Song , Yang Li , Fang Juemin , Wei Guochao , Ma Xiaochun , Zhao Zhikun , Weeraratne S Dilhan , Snowdon Jane , Wang Sheng , Qing Xu
Published: May 2019

e14300

Background: Acral and mucosal melanoma are rare subtypes accounting for about 3% of all melanoma cases. The cutaneous melanoma genomic landscape is well defined; however, little is known about the acral and mucosal melanoma mutational spectrum. In this pilot study, we evaluated the genomic and neo-antigen profiles and tumor mutational burden (TMB) from acral and mucosal melanoma patients with the aim of designing personalized vaccines and longitudinally tracking patients’ clinical courses. Methods: Tumor whole exome sequencing and neo-antigen profiling of 5 acral and 3 mucosal melanoma patients at Shanghai Tenth Peoples Hospital, Tongji University, China between April 2018 and January 2019 was performed using YuceBio’s proprietary analytics platform. Watsonä for Genomics, an artificial intelligence decision-support system, was used for variant interpretation and annotation. A comparative analysis was performed on Chinese acral melanoma data with the published Caucasian acral cohort from the Translational Genomics Research Institute (TGen) and The Cancer Genome Atlas (TCGA) predominantly Caucasian cutaneous melanoma data set. Results: TMB in our acral/mucosal melanoma cohort was 2.26/Megabase (Mb) compared to over 20/Mb in published cutaneous melanoma studies. Tumor neo-antigen burden (TNAB) in our group was 1.03 neo-epitopes/Mb. Low TNAB levels were associated with low TMB levels in all tumors. Incidence of BRAF and NRAS mutant cases in our cohort was 0% (0/8) and 13% (1/8) respectively compared to 19% (5/27) and 7% (2/27) of the Caucasian acral population in the TGen dataset. Incidence of BRAF and NRAS mutations in the TCGA cutaneous melanoma dataset was 54% (237/440) and 28% (125/440), respectively. Conclusions: TMB was significantly lower in acral/mucosal than in cutaneous melanoma and may be a surrogate for TNAB. Detection of BRAF and NRAS mutations, the two most prevalent driver mutations in cutaneous melanoma, were significantly lower frequencies in both Chinese and Caucasian acral melanoma patients in this study, suggesting alternate cancer drivers may exist in this subtype. Strategies to address challenges of low TNAB in vaccine development are being explored.
Effect of ligand binding domain activating mutations of ESR1 on cellular metabolism, liver tropism, and chemotherapy resistance.
Authors: Wolf Ido , Zinger Lotem , Merenbakh-Lamin Keren , Klein Goldberg Anat , Elazar Adi , Journo Shani , Boldes Tomer , Taya Marwa , Pasmanik-Chor Metsada , Spitzer Avishay , Rubinek Tami
Published: May 2019

e14506

Background: Mutations in the ligand binding domain (LBD) of estrogen receptor α (ER) confer constitutive transcriptional activity and resistance to endocrine therapies in breast cancer patients. Accumulating clinical data suggest adverse outcome for patients harboring tumors expressing these mutations. We aimed to elucidate mechanisms conferring this aggressive phenotype. Methods: Cells constitutively expressing physiologic levels of ER harboring activating LBD mutations were generated and characterized for viability, invasiveness and tumor formation in vivo. Gene expression profile was studied using RNAseq. Metabolic properties were assessed using global metabolite screen and direct measurement of metabolic activity. Response to chemotherapies was assessed using viability assays. Results: Cells expressing mutated ER showed increased proliferation, migration and in vivo tumorogenicity compared to cells expressing the WT-ER, even in the presence of estrogen. Experiments in mice revealed a more aggressive phenotype of the Y537S mutant compared to D538G as well as unique tissue predilection. Thus, 538G-ER cells exhibited liver tropism while Y537-ER cells mainly metastasized to the lungs and lymph nodes. Importantly, both mutations conferred resistance to paclitaxel and doxorubicin. Further studies indicated association of the mutated ER with upregulation of genes involved in tumor cell metabolism. Indeed, a global metabolic screen revealed distinct metabolic profile for cells harboring activating mutations, including the ability to utilize glutamine as an alternative carbon source. Moreover, we observed unique metabolic activities enabling cells to thrive in urea-reach environment of the liver, and overcome doxorubicin-induced stress responses. Conclusions: Taken together, these data indicate estrogen-independent rewiring of breast cancer cell metabolism by LBD-activating mutations, enabling aggressivene clinical behavior and chemotherapy resistance. Importantly, These unique metabolic activities may serve as a potential vulnerability and aid in the development of novel treatment strategies to overcome endocrine resistance.
Assessing the risk of chemotherapy toxicity and hospital admission due to toxicity.
Authors: Malton Samuel Robert
Published: May 2019

e14508

Background: Acute chemotherapy toxicity can have negative effects for the patient and the health economy. Finding overall toxicity incidence for a typical teaching hospital population proved difficult, although the 2008 report in to deaths within 30 days of chemotherapy found that 43% of patients who died reported a grade 3 or 4 toxicity1. At Nottingham University Hospitals (NUH) a Cancer Admissions and Triage Team was established, who undertake a proactive telephone toxicity assessment 24 hours following the administration of a first cycle of chemotherapy. This had the aim of reducing hospital admission due to toxicity through timely advice and intervention and allowed the accrual of data regarding reported toxicity. Methods: Data was obtained from the telephone assessment of acute toxicity 24 hours after administration of a first cycle of chemotherapy, once the service had been in use for 1 year. Descriptive statistics were produced to establish toxicity incidence and severity and hospital admission rates as well as length of stay. Regression modelling was used to identify predictors of the four outcome measures. The commonest toxicities were explored as secondary outcome measures. Results: 1539 patients were studied and the overall incidence of toxicity was 35.6%. Disease site and number of chemotherapy agents given were shown to predict toxicity, with breast and upper gastrointestinal cancers having a higher likelihood of toxicity. The more anticancer agents used, the higher the risk of toxicity. Disease was predictive of toxicity grade, with urology, gynaecology and lung patients experiencing higher grades of toxicity than other tumour sites. The rate of hospital admission due to toxicity was 13.1% and median length of stay 3 days. The risk of admission had some risk factors in common with toxicity. Disease and the number of drugs in the regimen affected the risk of admission, with gynaecology, head and neck and lung patients and patients who received 3 drugs having a higher likelihood of admission. No predictors could be reliably identified for length of stay. Predictors in the sub-groups of breast, lower gastrointestinal and lung cancers did not differ greatly from the whole population and the number of drugs was shown to be a predictor of nausea, vomiting and fatigue when explored as secondary outcomes. Conclusions: Predictors were identified for occurrence of toxicity, severity of toxicity, risk of hospital admission but not for length of stay. Sub-group analyses were undertaken. The overall burden of acute chemotherapy toxicity of a heterogeneous population was elucidated.
Rationale and preclinical development of LEAF-1401 and LEAF-1701: A novel class of potent next generation antifolates.
Authors: Moyo Victor M. , Xu Zhenghong , Khalifa Kaniz , Gandhi Nishant , Geng Bolin , Dhindsa Navreet , Niyikiza Clet
Published: May 2019

e14515

Background: Polyglutamation of antifolates by folylpolyglutamate synthase (FPGS) greatly enhances their activity, e.g. pentaglutamated pemetrexed is 80-fold more potent at inhibiting thymidylate synthase than pemetrexed. Polyglutamated antifolates however, do not readily cross the cell membrane, due to high negative charge and molecular mass. The resulting poor intracellular bioavailability greatly limits their therapeutic potential. Gamma glutamyl hydrolase (GGH) enzyme removes the glutamate residues from polyglutamates, which then are subject to efflux pumps. Resistance to pemetrexed has been linked to downregulation of both FPGS and folate transporters and upregulation of both GGH and the efflux pumps. We have been developing a novel class of nanoliposomal polyglutamated-antifolates including: LEAF-1401; nanoliposomal gamma-L pentaglutamated antifolate LEAF-1701; nanoliposomal alpha-L pentaglutamated antifolate These were designed to address key challenges of currently used antifolates namely: Bypassing the need for intracellular endogenous FPGS Direct delivery of the much more active polyglutamates through nanoliposome technology Minimizing toxicity to normal cells Mitigating against key antifolate resistance mechanisms such as downregulation of FPGS and folate transporters, and upregulation of GGH and efflux pumps Methods: In vitro testing in cell lines and in vivo testing in mice were performed using LEAF-1701 and LEAF-1401 in various tumor types. Results: Compared to pemetrexed: In vitro, LEAF-1701 and LEAF-1401 were more potent, in various cell lines while sparing normal neutrophils, colon and liver cells. In vivo, in H292 lung cancer xenografts, treatment with LEAF-1701 improved survival to 59 days vs 39 days for pemetrexed. In A549 lung cancer mouse orthotopic models, treatment with LEAF-1401 reduced metastatic tumor burden and prevented new metastases. In vivo, in mice doses of up to 80 mg/kg IV weekly were tolerated with little impact on blood counts and chemistry. Conclusions: LEAF-1701 and LEAF-1401 are innovative new chemical entities with promising preclinical activity and improved safety profiles and are now advancing to the clinic.
Integrin β5 in plasma-derived exosomal protein predicting liver-specific metastasis in advanced gastric cancer patients.
Authors: Li Qian , Xu Xiaojing , Yu Yiyi , Li Wei , Liang Li , Song Jia , Li Bin , Li Fu-Gen , Cai Shangli , Liu Tianshu
Published: May 2019

e14521

Background: Gastric cancer (GC) is the second leading cause of cancer death in China. Liver metastasis (LM) is a highly frequent event of the advanced GC, and the patients with LM always present dismal prognosis. A predicted factor of LM is urgently established to screen out the population in high risk of LM. Tumor-derived exosomes play critical roles in cancer metastasis process. Exosomes integrin (ITG) αvβ5 is uptake by liver resident cells and linked to LM in cell lines. This study is designed to evaluate the association between exosomes ITG αvβ5 and liver metastasis, especially in HER2 positive gastric cancer. Methods: 74 untreated advanced GC patients with matched blood and tumor tissue samples and clinical features were collected in the study. Exosomal HER2, ITG αv and β5 expression were measured by enzyme-linked immunosorbent assay (ELISA) after the exosomes isolated from plasma. HER2 status of tumor tissue was determined by IHC and/or FISH. Results: Based on HER2 status of tumor tissue, we initially proved the feasibility of detecting HER2 expression from exosome. ROC curves showed the optimum exosomal HER2 expression diagnostic cutoff for HER2 positive patients was 266.38 pg/ml (sensitivity 71.4% and specificity 72.6%). In total, 27 patients were LM and the rest (n = 47) was non-LM. The median expression value of ITG αv and β5 were 57.55 pg/ml and 836.00 pg/ml, respectively. And these two median value were identified as the cutoff value for subsequent analysis, respectively. HER2 expression was merely linked to ITG β5 expression (P = 0.019), not ITG αv (P > 0.050). There was no relationship between ITG αv and LM (P = 0.629), however, ITG β5 expression presented significant association with LM (P = 0.016). We further probed the relationship between ITG and other metastasis. Results showed no correlation between any ITG and other non-liver organic metastases, like lung, bone, ovarian, lymph, and peritoneal metastasis. Besides, ITG β5 expression was also associated with LM in the HER2-positive GC patients. Conclusions: Exosomes might serve as a potential non-invasive diagnostic and screening tool. In clinical samples, ITG β5, not αv, was linked to LM, and might be a valuable biomarker in the use of predicting LM of GC, even in the HER2-positive patients.
Imaging features of genomic instability in circulating tumor cells (CTCs) from metastatic triple-negative breast cancer (metTNBC) patients (pts) who received PIKTOR.
Authors: Ontiveros Priscilla , Levin Maren K. , Wang Yipeng , Hom Beverly , Dittamore Ryan , O'Shaughnessy Joyce
Published: May 2019

e14527

Background: A subset of TNBCs have homologous recombination deficiency (HRD) with upregulation of compensatory DNA repair pathways. A combination of TAK-228 and TAK-117 (PIKTOR), investigational oral TORC1/2 and PI3Kα selective inhibitors, respectively, is hypothesized to increase genomic instability (GI) and decrease double strand DNA repair. CTCs from metTNBC pts are being characterized for imaging features that may correlate GI in blood obtained at baseline (BL) prior to beginning PIKTOR, and at disease progression (PD) on PIKTOR. Pts are treated with a cisplatin-based regimen after PD on PIKTOR. Methods: Following IRB-approved informed consent, metTNBC pts received 4 mg PO TAK-228 and 200 mg PO TAK-117 QDx3d QW until PD. Blood samples collected at BL and at PD were sent to Epic Sciences for CTC analyses including enumeration, cell morphology, phenotypic heterogeneity, and GI analysis via a previously developed GI prediction algorithm based on cell phenotypes (Jendrisak et al AACR 2018). Results: The 9 pts treated with PIKTOR to date had mets in lymph nodes (n = 8), lung (n = 5), chest wall (n = 2), and bone (n = 2). Median number of prior regimens for metTNBC was 2; median duration of PIKTOR treatment was 8 wks (range, 5-15). CTCs were detected in 62.5% (5/8) BL and 100% (9/9) PD samples. 2 pts had high phenotypic CTC heterogeneity at both BL and PD (pt#36: 0.69 to 0.64; pt#80: 0.93 to 1.41), 1 pt had increased heterogeneity (pt#75: 0 to 1.68) and 1 pt had decreased heterogeneity (pt#67: 1.26 to 0) post-PIKTOR. 1 pt had predicted phenotypic GI (pGI+) CTCs detected at both BL and PD (pt#80), 1 pt had pGI+ CTCs at BL only (pt#36) and 1 pt at PD only (pt#75). Higher expression of the epithelial marker, cytokeratin (CK), and larger cell size were observed in the PD samples. Conclusions: Following treatment with and PD on PIKTOR, higher CK expression, higher phenotypic heterogeneity, and more pGI+ CTCs were observed in a subset of pts, suggesting that these TN CTCs may have developed epithelial transition and greater GI with PIKTOR treatment. Accrual continues and CTC GI biomarkers will be correlated with pts’ response to subsequent cisplatin-based therapy.
Modeling of pharmacodynamic (PD) biomarkers (BM) related to CDK4/6 inhibition and exploratory BM-response (progression free survival [PFS]) analyses in patients with advanced breast cancer.
Authors: Yu Yanke , Liu Yuan , Wang Diane Dan
Published: May 2019

e14528

Background: Palbociclib (PAL) is an oral inhibitor of cyclin-dependent kinase 4 and 6 (CDK4/6) approved for advanced breast cancer (ABC). In a clinical trial, the expression levels of PD BMs related to PAL effect, including thymidine kinase (TK) in serum and phosphor-Rb (pRb) and Ki67 in skin tissues, were measured at both baseline and post-treatment in patients with ABC treated with PAL plus letrozole (LET). Pharmacokinetic (PK)/PD modeling was conducted previously to characterize the longitudinal change of pRb and Ki67. The PK/PD was further evaluated for TK and exploratory analyses were conducted to evaluate the relationship between PFS and all three BMs. Methods: The present analyses used data from a Phase 1 study evaluating the PK, PD, safety and efficacy of PAL plus LET in 26 Chinese women with ABC. A 2 compartment model was used to describe the PK of PAL. A precursor-dependent indirect response model was developed to describe the TK time course after PAL treatment. PAL effect was modeled as Imax inhibitory model. Cox proportional hazard model was used to assess the correlation of PFS with three BM metrics (baseline BM, simulated lowest BM and maximum BM change in Cycle 1). Results: The BM data included 194 TK observations from 26 patients. The PK/PD models adequately described the longitudinal change of TK, and showed PAL caused TK reduction. The estimated IC50 value was 49.5 ng/mL, similar to those for pRb and Ki67. The BM-response analyses for PFS showed that correlation was found for TK. There was a significant correlation between PFS and baseline TK (BTK) and minimum TK (MTK) in Cycle 1. A longer PFS was associated with lower BTK and lower MTK. A trend for longer PFS with higher maximum TK change in Cycle 1 was observed although the relationship was not statistically significant. Conclusions: PAL exposure had significant correlation with the reduction of all three BMs. Longer PFS was associated with lower BTK and MTK. Due to the small sample size (N = 26), this analyses result need to be confirmed in a larger study.
Inflammatory biomarkers as pronostic factors of mortality in critical ILL oncology patients at ICU.
Authors: Aliaga Macha Carlos , Runciman Thanya , Carracedo Carlos F.
Published: May 2019

e14535

Background: Inflammatory markers have been used as prognostic factors in multiple malignancies.In cancer patients, critically ill, the utility of these have limited data.The aim of our study is to determine whether neutrophil lymphocyte ratio (NLR) or lymphocyte platelet ratio(PLR) are prognostic factors for mortality in critically ill patients. Methods: We retrospectively analyzed data of 79 patients with solid tumors admitted to ICU at Sanna-Aliada Clinic between January 2018 to December 2018. Inflammatory markers results were obtained from laboratory tests performed during the first 24h of admission to ICU. Receiving operating characteristic (ROC) curves were constructed and the sensitivity, specificity, predictive values and probability indicators for the NLR and PLR. Results: A total of 79 patients were assessed, 39 women and 40 men. The average age was 60.28 years, median of 61 ( 18 to 91). 51.9% had metastatic disease. The most frequent places were lung 12 (15.2 %) and brain 9 (11,4%) . The main cause for admission to ICU was infectious disease (40.5%). The analysis of normality (Kolmogorov-Smirnov test) indicates that the variables age, hemoglobin, leukocytes, platelets, neutrophils, lymphocytes, have a normal deviation while the other variables: lactate, PCR, neutrophil to lymphocyte ratio (NLR) , Platelet to lymphocyte ratio (PLR) are not distributed normally. Regarding mortality, 44 patients were alive at 30 days (66.7%), and 30 (45.5%) were alive at 90 days. The average stay in the ICU was 8.43 days, with a median of 6, (SD 7.17, 1 to 40 days), 22.8% died in the ICU. The evaluation of PLR and NLR as a mortality marker is significant for the group of patients admitted to the ICU due to a noninfectious pathology, generating an area under the curve (AUC) of 0.706 for NLR (95% CI, 0.535 - 0.876, p-value = 0.035) and 0.767 for PLR (95% CI, 0.615-0.918; p-value = 0.006); the optimal cut point by Youden’s index for NLR was 8.29 and 267.94 for PLR (Sensitivity: 76%, Specificity: 67%). In contrast, the group with infectious pathology, the AUC was 0.47 for NLR (p = 0.78) and 0.42 for PLR (p = 0.44). The relationship of the biomarkers with stay in ICU was also evaluated, finding a statistically significant association with the lactate value (p = 0.024, Kruskal-Wallis) Conclusions: Inflammatory markers are useful as predictive markers of mortality in critically ill patients due to non-infectious causes. The lactate value serves as a predictive factor of stay in the ICU for all the patients. We suggest carrying out prospective studies to confirm the validity of our findings.
The utility of circulating tumor DNA analysis to identify BRAF mutations in malignant melanoma.
Authors: Kirby Eric , Leming Philip D.
Published: May 2019

e14539

Background: Circulating tumor DNA provides an opportunity to assess the genetic profile of a patient’s tumor without performing a tissue biopsy. The Guardant360 assay is one platform available to perform genetic sequencing of circulating tumor DNA from a blood sample. This can allow for assessing the BRAF mutation status of patients with malignant melanoma from a liquid biopsy. Methods: Retrospective chart review was done of patients with malignant melanoma with a positive BRAF V600 mutation based on a tissue biopsy. The BRAF V600 mutation status of these patents as determined by a Guardant360 assay, which was not necessarily done at the same time as the tissue biopsy, was gathered. The status of each patient’s malignancy at the time of each liquid biopsy was assessed and divided into three categories, which included active distant metastatic disease, active locoregional disease, and no evidence of active disease. Results: 51 total patients with a positive BRAF V600 mutation based on tissue biopsy were included in this analysis. Some patients had multiple Guardant360 assays which were done at different timepoints, which made for 83 total liquid biopsies. Out of 30 liquid biopsies done when the patient had evidence of active distant metastatic disease, 21 (70%) were positive for a BRAF V600 mutation. Out of 13 liquid biopsies done when the patient had evidence of active locoregional disease, 2 (15.4%) were positive for a BRAF V600 mutation. Out of 40 liquid biopsies done when the patient had no evidence of active disease, 3 (7.5%) were positive for a BRAF V600 mutation. Conclusions: The Guardant360 assay in detecting a BRAF V600 mutation had it’s highest sensitivity in the setting of active distant metastatic disease. This assay can allow for sparing patients more invasive tissue biopsies when the suspicion for metastatic melanoma is high and the BRAF mutation status is unknown.
REG4 as a novel biomarker of lung adenocarcinoma with mutated KRAS and TTF-1 low expression.
Authors: Yu Hui , Sun Si
Published: May 2019

e14541

Background: Recent research has classified lung adenocarcinoma patients with KRAS mutation into three subtypes by co-occurring genetic events in TP53 (KP subgroup), STK11/LKB1 (KL subgroup) and CDKN2A/B inactivation coupled with low expression of the TTF-1 (KC subgroup). The aim of this study was to identify novel biomarkers through searching the candidate molecules that contributing to lung adenocarcinoma pathogenesis, especially KC subtype. Methods: We analyzed the publicly available database and identified the candidate REG4 using the E-GEOD-31210 dataset, and then confirmed by TCGA dataset. In addition, an independent cohort of 55 clinical samples was analyzed by quantitative real-time PCR analysis. Functional studies and RNA sequencing were performed after the silencing REG4 expression. Results: REG4, an important regulator of gastro-intestinal carcinogenesis, was highly expressed in KRAS mutated lung adenocarcinoma with low expression of TTF-1 (KC subtype). The results were validated both by gene expression analysis and immunohistochemistry study in an independent 55 clinical samples from Fudan University Shanghai Cancer Center. Further in vitro and in vivo functional assays revealed silencing REG4 expression significantly reduce cancer cell proliferation and tumorigenesis. Moreover, RNA sequencing and GSEA analysis displayed that REG4 knockdown might induce the cell cycle arrest by regulating G2/M checkpoint and E2F targets. Conclusions: Our results indicate that REG4 plays an important role in KRAS-driven lung cancer pathogenesis and is a novel biomarker of lung adenocarcinoma subtype. Future studies are required to clarify the underlying mechanism of REG4 in the division and proliferation of KC tumors and its potential therapeutic value.
Real-world application of liquid biopsy in gastrointestinal tumors: Experience from a comprehensive cancer center.
Authors: Sanz-Garcia Enrique , Rodriguez Laura , Alvarez-Gallego Rafael , Rodriguez-Pascual Jesus , Munoz Cesar , Ugidos Lisardo , Ceniceros Lucia , Boni Valentina , De Miguel Maria José , Almirall Cristina , Calvo Emiliano , Ayuso-Sacido Angel , Cubillo Gracián Antonio
Published: May 2019

e14546

Background: Determination of circulating tumoral DNA (ctDNA) in plasma has recently emerged as an alternative to genomic testing in solid biopsy. Although different clínical trials have suggested that ctDNA alterations can tailor patient's treatment, its routinary utility in clinical practice is still controversial. Methods: A total of 74 consecutive plasma samples from 63 patients (pts) have been collected from October 2017 to December 2018 in our center and analyzed using two different next-generation sequencing (NGS) platforms: OncoTRACE (OT) and Guardant360 (G360). Main epidemiological, pathological and clinical data have been retrospectively extracted from medical records and correlation with ctDNA alterations have been performed using Chi-Square test. Application of liquid biopsy results has been decided after discussion in molecular tumor board and physician’s opinion. Results: OT and G360 were performed in 63 and 11 plasma samples respectively. Most part of the samples were from pancreatic (34; 45.9%) and colorrectal (CRC) (28; 25.7%) adenocarcinoma pts although biliary tract, gastric and oesophageal carcinoma pts were also included (4 each; 5.4%). Plasma samples were analyzed after progression to ≥two systemic therapies in 48.6%. At least one pathogenic alteration of ctDNA was detected in 44.6% of the samples. No correlation has been observed between presence of alteration in ctDNA and tumor origin, surgery of the primary, number and location of metastases and CEA and CA19.9. ctDNA alteration could be targetable with approved or experimental therapies in 14 pts (20.9%; 6 pts OT and 8 pts G360): 9 pts with CRC ( BRAF, EGFR, RAS, PI3KCA, PTEN), 2pts with pancreatic ( IDH1, BRCA1) and 1 pt with gastric, oesophageal and biliary tract carcinoma ( PTEN, EZH2, IDH1). However, these results have been applied in 4 pts (6% of all pts, 28.6% with targeteable ctDNA alterations): 3 pts were treatment-naïve and 1 pt had progressive disease to 2 systemic therapies. Most part of the patients with targetable alterations who did not receive treatment was due to clinical deterioration. Conclusions: Detection of alterations in ctDNA using NGS platforms is feasible in gastrointestinal tumor patients and can help physicians to decide treatment, especially in CRC. However, many patients could not be treated according to ctDNA as plasma samples were collected in late stages of the disease.
Analysis of AKR1B10 levels among various groups of breast cancer patients.
Authors: Rao Krishna A. , Robinson Kathy , Delfino Kristin , Tiwari Vivekanand , Zhu Yun , Wang Xin , McClintock Andrew , Bekker Simon , Westbrook Joshua , Cao Deliang , Cao Zhe
Published: May 2019

e14548

Background: Aldo-keto reductase 1B10 (AKR1B10) is a protein that is primarily expressed in human colon and small intestine, but induced in hepatocellular carcinoma and non-small cell lung cancer. Our recent studies have revealed that AKR1B10 is overexpressed in primary, metastatic, and recurrent cancers of the breast.

Methods: We recruited four cohorts of patients: Patients with breast cancer undergoing primary surgery from whom we procured breast cancer and matched normal adjacent tissue to evaluate AKR1B10 expression in primary tumors. The matched serum samples were collected before surgery and at various time points after the surgery (approximately 3 days, 7 days, and 1 month Patients with recurrent or advanced (metastatic) breast cancer. Serum samples were collected and serially monitored for up to 2years before and during metastatic therapy and correlated with RECIST measurements. Patients with locally advanced disease undergoing neoadjuvant chemotherapy. AKR1B10 serum levels were monitored during therapy and correlated with RECIST measurements. Healthy individuals with normal mammograms were recruited and submitted serum samples for AKR1B10 serum measurements. Results: AKR1B10 rapidly cleared the serum of early stage breast cancer patients with an estimated half-life of 24-30 hours. Serum AKR1B10 levels correlated strongly with tissue IHC staining and PR positivity but not with RECIST levels or Oncotype Dx scores. Neoadjuvant chemotherapy did not affect serum AKR1B10 levels. Individuals with normal mammograms displayed substantially lower levels of AKR1B10 than breast cancer patients. Patients with DCIS also displayed elevated serum AKR1B10 levels.

Conclusions: AKR10 may serve as tumor marker to independently identify high risk patients whose tumors have intermediated risk Oncotype Dx scores and may also identify early breast cancers in patients with equivocal breast biopsies.
Detection of circulating cancer stem cells (cCSCs) as a predictive biomarker for breast cancer relapse or metastasis.
Authors: Pizon Monika , Pachmann Ulrich A. , Pachmann Katharina , Schott Dorothea
Published: May 2019

e14559

Background: Breast cancer is one of the most common types of cancer in women worldwide. It has been demonstrated that even localized tumors without clinically apparent metastases give rise to circulating epithelial tumor cells (CETCs). Recent studies have provided strong support for the cCSC hypothesis, which suggests that a more aggressive subpopulation of circulating tumor cells is the source of metastatic spread from the primary tumor. Measuring CETCs in blood of patients has emerged as a non-invasive diagnostic procedure for screening patients who may be at high risk for developing metastatic cancers. However, accurate detection of CETCs may provide erroneous results, since CETCs undergoing EMT during metastasis are down-regulated for the expression of epithelial cell markers. Methods: 26 breast cancer patients were included into the study. The determination of CETCs and cCSC was performed using maintrac method and tumorsphere forming assay, respectively. Cell viability, surface marker expression and ALDH 1 activity of the cells in the spheres were evaluated by fluorescence scanning microscope. Results: Sphere formation was observed in 80 % of patients. We found that the number of tumorspheres depended on stage of disease. The number of tumorspheres increased significantly with tumor progression, especially with the presence of metastases. Tumorsphere formation was observed in all metastatic patients (median of 30 tumorspheres/100µl of blood), although only 27% of them had detectable CETCs. Analysis of surface marker expression profile showed that the cells in the spheres had typical phenotype of cancer stem cells. Sphere formation was not observed in healthy subjects (n = 20). Conclusions: There is a high correlation between the numbers of tumorspheres cultured from peripheral blood with clinical stage of disease. Identifying cCSC in blood sample can help clinicians to monitor breast cancer patients and to detect metastasis in early stages.
Use of a clinical trials screening tool in the NCI Community Oncology Research Program (NCORP) to enhance accrual and promote disparities research.
Authors: St. Germain Diane C. , Budd Troy , McCaskill-Stevens Worta J.
Published: May 2019

e14587

Background: Despite the identification of multiple patient, clinician, and health systems barriers, accrual to a new generation of cancer clinical trials continues to present challenges. It may be that barrier data is not sufficiently granular. A clinical trial screening tool was developed for use in NCORP to collect trial- and site-specific information as well as broadened demographic data to determine factors that may impact accrual. Methods: The tool was developed with stakeholder input and the NCI Oncology Patient Enrollment Network was used for data input and analyses. Results: From February 2016 to December 2019, 14,340 entries were made in the screening tool. Eighty-two percent of participants consented to participate. Participants screened were female (77%), married (64%) and Caucasian (85%). Fourteen percent of the participants were racial minorities (1% not reported) and 5% were Hispanic or Latino. The mean age was 60 (range 1-95). Thirty-six percent were employed, ≥ 32 hours per week followed by 35% retirees. Income did not vary significantly ( < $25,000(16%), $25,000 - $50,00 (20%), $51,000 - $10,000 (26%), and > $100,000 (19%), and 19% of participants refused to provide income data. Four percent of the participants were uninsured at diagnosis. Seventy-two percent (8,501) of participants screened enrolled in a clinical trial. Of those not enrolled, 49% were ineligible and 48% were eligible but declined to participate. The most common reasons for ineligibility included concurrent disease, abnormal lab or other tests, and patient could not comply with eligibility criteria. The most common reasons eligible participants declined to participate were perception that toxicities were too great and social issues (child care, transportation). Further analysis of the data will include correlation of race/ethnicity, age, income and co-morbidities with enrollment status. Conclusions: The majority of participants approached agreed to participate in the screening tool protocol. Approximately half of the participants were eligible for a trial but declined to participate. These issues will be addressed within the network to enhance accrual. The data collected will also provide opportunities for investigators within the network to develop research questions focused on disparities and clinical trial accrual.