bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of biopsy methodology on biomarker testing in NSCLC in EU5.
Authors: Homer Christophe
Published: May 2019

e14264

Background: With the increasing number of drug treatments in NSCLC reliant on complementary or companion diagnostics and the evolution of testing methodologies that require larger sample volumes, biopsy remains a gatekeeper to providing adequate sample for biomarker testing and informing treatment decisions. Methods: The Ipsos Global MDx Monitor 2018 Q3 NSCLC EU5 (UK, France, Germany, Italy, Spain) Module: an online multi-stakeholder study involving drug treating physicians (225) and pathologists (81) who answer perceptual questions and report de-identified medical chart data on NSCLC patients (1257) / samples (516), that have undergone at least 1 biomarker test. Study fielded July-September 2018. This abstract reviews pathologist data only. Results: EU5 pathologists (n = 81) state that in 14% NSCLC cases there was not enough sample to carry out any biomarker tests, in 19% cases there was enough sample to carry out some requested biomarker tests and in 67% cases there was enough sample to carry out all biomarker tests. On analyzing NSCLC sample record forms, 83% (421/505) samples from unique patients were from solid samples (FFPE block /Fresh frozen), 14% liquid samples (fluid/cytology) (73/505). Of the 487 NSCLC sample records submitted, 39% biopsies were performed by bronchoscopy (190/487), 25% by Transthoracic Core Needle Biopsy (CNB) (126/487), 16% by Transthoracic Fine Needle Aspiration (FNA) (78/487), 8% by Open biopsy/resection (39/487). Sample sufficiency varies by biopsy methodology; 100% open biopsies/resection (39/39) provided enough sample to carry out all tests ordered - versus 87% for Transthoracic CNB (99/113), 84% for bronchoscopies (155/184), 67% for Transthoracic FNA (49/73). Sample sufficiency varies by sample type; 89% solid samples (357/399) provided enough sample to carry out all tests ordered, 75% liquid samples (49/65). Conclusions: Sample sufficiency and thus ability to test for biomarkers is affected by the biopsy method used to extract the sample, and sample type used for testing (i.e. liquid vs. solid). Sample insufficiency can necessitate a re-biopsy or lead to treatment decisions being taken without the tumour’s full genomic profile at the physician’s disposal.
Coexpression patterns of IDO-1, PD-L1 and EGFR in non-small cell lung cancer.
Authors: Wang Weili , Zhang Chen , Cheng Liang , Jin Jianyue , Kong Feng-Ming Spring
Published: May 2019

e14279

Background: Indoleamine-2, 3-dioxygenase (IDO), a well-established immune suppressor, may offset efficacy of immune checkpoint inhibitors (ICIs) such as programmed death receptor ligand-1(PD-L1) by inducing unfavorable tumor microenvironment. The unpredictability of one ICI response may partially due to the concomitant presence of other ICIs. Therefore, increasing interest has been focus on the dual ICIs or the combination of ICIs with conventional treatment modalities such as radiotherapy, chemotherapy as well as target therapy. This study aimed to study the coexpression patterns and significance of IDO-1, PD-L1 and epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC). Methods: Patients with newly diagnosed NSCLC enrolled in three prospective surgery clinical trials were included in this study. The expressions of IDO-1, PD-L1 and EGFR were evaluated by immunohistochemistry (IHC). For positive cases, the percentile score and H-score were generated respectively from two independent pathologists. The descriptive analysis of Crosstab was used to compare the distributions of these biomarkers expression by pathological types. Results: A total of 117 patients (adenocarcinoma -54%, squamous cell carcinoma -32% and others -14%) with pretreatment tissue specimen available for IHC analysis were studied. Of these, 105 (90%) were from lung primary and 12 (10%) from distant metastasis sites. The expressions of IDO-1 (≥ 1%), PD-L1 ((≥ 1%) and EGFR (≥ 90% or intensity = 3) were identified in 43%, 40% and 52% of NSCLC patients. For IDO-1 and PD-L1, coexpression rates were 21% for co-positive and 37% for co-negative. However, there were still 42% patients showed isolated PD-L1 (20%) or IDO-1 expression (22%). The coexpression rates for EGFR and IDO-1 were 21% for co-positive and 26% for co-negative. The coexpression rates for EGFR and PD-L1 were 20% for co-positive and 25% for co-negative. 38% patients had a positive EGFR with either IDO-1 or PD-L1 positive. The expression patterns were not significant associated with clinical factors including pathological types and differentiation grades. Conclusions: IDO-1 and PD-L1 expression patterns may be useful in the selection of NSCLC patients for dual ICIs immunotherapy. The combinations of EGFR target therapy with immunotherapy may benefit some of NSCLC patients. However, testing expression status of EGFR and immune checkpoints becomes essential since majority of NSCLC patients exhibits EGFR positive with IDO-1 or PD-L1 negative patterns.
Breast MRI as a radiomic biomarker of immune enrichment in HER2+ breast cancer.
Authors: Kennedy Laura , Durenberger Grace , Stanton Sasha E. , Vinayak Shaveta , Dintzis Suzanne , Partridge Savannah C. , Gadi Vijayakrishna K.
Published: May 2019

e14281

Background: Trastuzumab is an important drug in the treatment (tx) of HER2+ breast cancer, but not all tumors are responsive. Immune-enriched tumors have a better response to tx, and a biomarker to identify these tumors would be valuable. Multiparametric functional MRI is a non-invasive technique that can assess the entire tumor without the sampling error of biopsy. In this pilot study, we evaluate breast MRI as a potential biomarker for tumor immune response to trastuzumab. Methods: Women with de novo cT1-2N0 HER2+ breast cancer were enrolled and received paired multiparametric breast MRI examinations (including DWI and DCE sequences) prior to and ~2 weeks after a single dose of trastuzumab, followed by surgery or biopsy (if additional neoadjuvant tx planned). Tumor peak signal enhancement ratio (SER) and mean apparent diffusion coefficient (ADC), reflecting tissue perfusion and tissue cellularity, were determined from the images. Pre- and post-tx tissue were immunologically profiled with TIL counts and an immune-focused RNA gene expression panel. SER and ADC were correlated with TIL and RNA gene expression using Spearman’s rho. Results: To date, 8 women have enrolled and 6 had pre- and post-tx tissue available. The primary endpoint was correlation between SER and ADC measurements with tumor TILs. Secondary endpoints included correlating SER with angiogenesis and inflammation markers and correlating changes in SER and ADC with apoptosis/cell death markers. About half of the patients had TIL-enriched (TIL≥ 50%) tumors at baseline, and after trastuzumab the majority of tumors remained enriched or had an increase in TILs. Results suggest that pre-tx ADC had a negative correlation with TILs by histology and gene expression, although correlations did not reach statistical significance in this small cohort. Results also suggest a strong correlation between pre-tx SER and VEGF-A (r = 0.83) with a trend towards significance (P = 0.10). A decrease in SER post-tx had a strong correlation with an increase in caspase 3 expression (r = -0.85, p = 0.05), which suggested tx effect. Conclusions: Functional MRI may provide a novel biomarker of breast tumor immune response to trastuzumab. Results from additional patients will be available at the meeting.
Frequency of discordance in programmed death ligand 1 (PD-L1) expression between primary tumors and paired distant metastases in advanced cancers: A systematic review and meta-analysis.
Authors: Lee Chia Ching , Tham Ivan Weng Keong , Tan Char Loo , Lum Jeffrey , Tey Jeremy Chee Seong , Soon Yu Yang
Published: May 2019

e14290

Background: Randomized trials have demonstrated that PD-L1 expression in tumor cells predicts for response to PD-1 or PD-L1 checkpoint inhibitors in advanced cancers. However, the frequency of discordance in PD-L1 expression between primary tumors and paired distant metastases is currently unclear. We aimed to determine the discordance rate of PD-L1 expression between primary tumor and paired distant metastases in advanced cancers. Methods: We searched MEDLINE and EMBASE for eligible studies. We noted the discordance rate (positive-to-negative or vice versa) and performed subgroup analyses based on the PD-L1 expression in primary tumor, locations of primary tumor and distant metastases, positivity thresholds and type of antibody used for testing. We used QUADAS-2 tool to assess the methodologic quality of the included studies. We performed the meta-analysis using the logistic-normal random effects model. Results: We identified 11 eligible studies including 351 cases of lung, colorectal, breast and ovarian cancers. Nine studies were judged to have low risk of bias in their methodological quality. The overall discordance rate in PD-L1 expression between primary tumor and paired distant metastases was 28% (95% confidence interval (CI) = 18-41) with significant heterogeneity among the studies (chi-square test P-value < 0.0001). There was no statistically significant differences in discordance rates between the subgroups: PD-L1-positive (40%, 95% CI = 21-63) vs PD-L1-negative primary tumors (21%, 95% CI = 12-34), lung primary (31%, 95% CI = 14-55) vs colorectal/ breast/ ovarian primary (27%, 95% CI = 19-37), central nervous system metastases (22%, 95% CI = 8-47) vs liver/ lung/ pleural/ peritoneum metastases (34%, 95% CI = 24-46); 1% (23%, 95% CI 16-33) vs 5% positivity threshold (33%, 95% CI 11-68); E1L3N (42%, 95% CI 10-82) vs SP142 antibody (27%, 95% CI 20-36). Conclusions: The discordance in PD-L1 expression between primary tumors and paired distant metastases is close to 30%, which could potentially affect response to immunotherapy. Further works exploring the mechanisms and impact of this discordance are warranted.
Systematic assessment of LCMV based vaccine vectors expressing melanocyte differentiation antigens in human in vitro assays and in mouse melanoma models.
Authors: Flatz Lukas , Cupovic Jovana , Schmidt Sarah , Onder Lucas , Abdou Marie Therese , Fritsche Anna Katharina , Bonilla Weldy , Pop Oltin Tiberiu , Tüting Thomas , Bergthaler Andreas , Orlinger Klaus , Bald Tobias , Ludewig Burkhard , Ring Sandra
Published: May 2019

e14299

Background: Antibodies blocking the immune checkpoint pathways represent an important milestone in the treatment of patients with metastatic melanoma. However, individuals presenting cold tumors do usually not show a clinical benefit. New cancer vaccine approaches are needed to cover this gap. Methods: A novel replication attenuated vaccine vector based on lymphocytic choriomeningitis virus expressing full length melanocyte differentiation antigens is evaluated in mouse melanoma models and in PBMCs and T cell cultures from melanoma patients. Results: Here, we demonstrate that intratumoral but not intravenous injection of a recombinant propagating LCMV vector expressing the melanoma-associated antigen TRP2 leads to T cell-dependent eradication of established s.c. melanoma. Importantly, intratumoral vaccination shows an abscopal effect on distant lung metastasis and protects from a rechallenge with melanoma. Confocal microscopy and flow cytometry reveal that intratumoral injection of rLCMV vectors reprograms the tumor microenvironment resulting in sustained T cell fitness. In addition, we demonstrate that rLCMV vectors can efficiently transduce human antigen presenting cells. Moreover, in vitro data confirm that rLCMV efficiently induces T cells against various melanoma-associated antigens in PBMCs from melanoma patients. Conclusions: Preclinical assessment of propagating rLCMV vectors shows unique features of this cancer vaccine resulting in a profound and multistep activation of the cancer immunity cycle resulting in eradication of established melanomas in the B16F10 mouse model after one single immunization. Positive proof of principle experiments using PBMCs from melanoma patients suggest a rapid evaluation in clinical trials.
Live-attenuated LCMV-based vector for active immunotherapy of HPV16+ cancer.
Authors: Schmidt Sarah , Bonilla Weldy V. , Pauzuolis Mindaugas , Reiter Andrea , Kleissner Theresa , Oeler Daniel , Stemeseder Felix , Berka Ursula , Kiefmann Bettina , Schulha Sophie , Matushansky Igor , Merkler Doron , Pinschewer Daniel , Orlinger Klaus Karl
Published: May 2019

e14303

Background: Active immunization against cancer requires the induction of exquisitely potent tumor-specific CD8+ T cell (CTL) responses, which can be repeatedly boosted and reactivated. Hookipa Pharma engineered a replication-attenuated viral vector platform (TheraT) based on the arenavirus lymphocytic choriomeningitis virus (LCMV), which meets these aforementioned criteria. Here, we present a preclinical data package of Hookipa´s lead immunotherapy product HB-201, targeting HPV16-driven cancer. Methods: TheraT-E7E6 encodes a highly immunogenic, non-oncogenic version of the human papilloma virus 16 (HPV16) oncoproteins E7 and E6. Results: Attenuation and safety of TheraT-E7E6 were demonstrated by i) rapid viral clearance after systemic administration of the vector and ii) reduced neurovirulence in mice.TheraT-E7E6 can be administered intravenously as systemic therapy or intratumorally as local therapy with an abscopal-like effect. Treatment was shown to induce substantial CD8+ T cell expansion and high frequencies of E7- and E6-specific CTL responses with a balanced effector / central memory profile. These responses were boosted and reactivated upon TheraT-E7E6 re-administration. TheraT-E7E6 eradicated palpable tumors in a syngeneic mouse TC-1 tumor model of HPV-driven cancer. Even in mice with large tumors (~300mm3) TheraT-E7E6 afforded significant tumor control and improved survival, with high frequencies of E7-specific CTLs persisting for several weeks. Animals which cleared the tumor after TheraT therapy were long term protected from tumor re-challenge. Furthermore, TheraT is efficacious in checkpoint inhibitory refractory models and can also synergize in combination with checkpoint inhibitors (CPIs). Conclusions: In conclusion, replication-attenuated TheraT-E7E6 is safe, highly immunogenic and shows excellent therapeutic efficacy as mono- or combination therapy in a preclinical model of HPV-induced cancer. Clinical trials for the treatment of HPV16+ cancers will be initiated in 2019.
A phase I study of the safety and immunogenicity of a multi-peptide personalized genomic vaccine in the adjuvant treatment of solid tumors and hematological malignancies.
Authors: Blazquez Ana , Rubinsteyn Alex , Kodysh Julia , Finnigan John Patrick , Marron Thomas Urban , Meseck Marcia , O'Donnell Timothy , Hammerbacher Jeffrey , Donovan Michael J. , Mahajan Milind , Miles Brett A. , Irie Hanna , Tiersten Amy , Tewari Ashutosh , Parekh Samir S. , Nair Sujit , Galsky Matt D. , Schadt Eric E. , Friedlander Philip Adam , Bhardwaj Nina
Published: May 2019

e14307

Background: Mutation-derived tumor antigens (MTAs) arise as a direct result of somatic variations that occur during carcinogenesis and can be characterized via genetic sequencing and used to identify MTAs. We developed a platform for a fully-personalized MTA-based vaccine in the adjuvant treatment of solid and hematological malignancies. Methods: This is a single-arm, open label, proof-of-concept phase I study designed to test the safety and immunogenicity of Personalized Genomic Vaccine 001 (PGV001) that targets up to 10 predicted personal tumor neoantigens based on patient’s HLA profile (ClinicalTrials.gov: NCT02721043). Results: Patients who completed vaccination with PGV001_002 (head and neck squamous cell cancer) received 10 doses of vaccine comprising 10 long peptides (LP) combined with poly-ICLC (toll-like receptor-3 agonist) intradermally. Vaccine-induced T-cell responses were determined at weeks 0 and 27 (before and after treatment, respectively), ex vivo by interferon (IFN)-g enzyme-linked immunospot assay and after expansion by intracellular cytokine staining. Overlapping 15-mer peptides (OLPs) spanning the entirety of each LP and 9-10-mer peptides corresponding to each predicted class I epitope (Min) were pooled. Ex vivo responses to these peptide pools were undetectable at week 0 but were evident at week 27 against 2 OLPs out of 10 (20%) and in 5 Min out of 10 (50%). After in vitro expansion, neoantigen-specific CD4+ and CD8+ T-cell responses were found in 5 out of 10 pooled peptides (50%). 7 out of 10 (70%) epitopes elicited polyfunctional T-cell responses (secretion of INF-g, TNF-a, and/or IL-2) from either CD4+ or CD8+ T cells. Conclusions: The PGV001 vaccine in our first patient showed both safety and immunogenicity, eliciting CD4+ and CD8+ responses to the vaccine peptides. As we enroll additional patients in this clinical trial, and perform deeper phenotyping of their tumor-reactive T cells, we will learn the determinants necessary for the successful generation of MTA-based vaccines, while informing future immunotherapeutic approaches and rational combinations. Clinical trial information: NCT02721043.
Role of metabolic reprogramming toward fatty acid oxidation on tumor lymph node metastasis.
Authors: Lee Choong-kun , Jeong Seung-hwan , Jang Cholsoon , Bae Hosung , Kim Yoo Hyung , Park Intae , Kim Sang Kyum , Koh Gou Young
Published: May 2019

e14501

Background: Metastasis of tumors to lymph node (LN) is a strong predictor of cancer patient mortality and determinant of disease progression and treatment options. Despite the clinical significance of tumor LN metastasis, the mechanisms underlying this process remain largely unexplored. Methods: We used comparative transcriptomics and metabolomics analyses of the primary tumor and LN micro- and macro-metastatic tumors from B16F10 murine melanoma model. B16F10 melanoma footpad implantation model and MMTV-PyMT spontaneous breast cancer genetic mouse models were analyzed in vivo. We also investigated the LN-metastatic tumors from sentinel LNs in 21 patients with melanoma. Results: We found that tumor cells undergo dramatic metabolic reprogramming towards fatty acid oxidation (FAO) for successful LN metastasis. Mechanistically, transcription co-activator YAP is selectively activated in LN metastatic tumors, leading to stimulation of FAO gene programs. Pharmacological inhibition of FAO or genetic ablation of YAP in tumors markedly suppressed LN metastasis. Unexpectedly, metastatic tumors highly stimulated the bile acid synthesis pathway upon arrival at the LN, and these bile acids activated YAP of tumor cells via nuclear vitamin D receptor. Lastly, YAP activation in the metastatic LNs of melanoma patients was reversely correlated with distant metastasis–free survival. Conclusions: This study propose that inhibition of FAO or YAP signaling to prevent the metabolic reprogramming of metastatic tumors offers a potential therapeutic strategy for mitigating tumor LN metastasis.
The relationship between mesenchymal stem cells, monocyte-macrophages, endothelial progenitor cells, and prognosis of hepatocellular carcinoma.
Authors: Yan Kun
Published: May 2019

e14507

Background: This experiment is designed to investigate the clinical correlation between mesenchymal stem cells, macrophages, endothelial progenitor cells and hepatocellular carcinoma, furthermore to assess their diagnostic value. Methods: A prospective cohort study was conducted to collect patients who admitted to the Department of General Surgery, Second Affiliated Hospital of Xi'an Jiaotong University. The peripheral blood samples of patients with hepatocellular carcinoma (HCC) who diagnosed by pathology from September 2014 to August 2016 were collected. The mesenchymal stem cells, macrophages and endothelial progenitor cells (EPCs) were analyzed by flow cytometers. Results: There was no correlation between hepatocellular carcinoma and peripheral blood mesenchymal stem cells (p > 0.05), and there was correlation between hepatocellular carcinoma and macrophages and endothelial progenitor cells (p < 0.01). The positive correlated coefficient is 0.938 in macrophages, and the negative correlated coefficient is -0.835 in the endothelial progenitor cells. Conclusions: The differentiated pathways of mesenchymal stem cells in hepatocellular carcinoma play key roles in the progression and prognosis of hepatocellular carcinoma.
A breakthrough in treatment of cancer patients with grade 4/profound febrile neutropenia.
Authors: Cainap Calin Ioan , Cainap Simona Sorana , Cetean- Gheorghe Sanziana , Leucuta Daniel Corneliu , Bochis Ovidiu Vasile , Gherman Alexandra
Published: May 2019

e14513

Background: febrile neutropenia (FN) remains one of the most serious side effects of chemotherapy treatment, which is life-threatening. Late consequence of FN is dose reduction and delaying of therapies. Actual guidelines allows granulocyte-colony-stimulating factors (G-CSF) in management of profound FN in addition to antibiotics and supportive care Methods: it is an original prospective study, approved by local ethics comitee, which included 73 patients with confirmed malignancy, treated in our Institute with chemotherapy, who developed febrile neutropenia (FN) and were hospitalised. Patients with myeloablative chemotherapy and bone marrow transplantation support, severe renal impairment, abnormal liver function or with a history of allergic reactions to the antibiotics were excluded from the study. There were recorded 96 episodes of grade 4 FN (1-3 episodes / patients). Each of one was hospitalised according to institutional guideline but with a dose of G-CSF of 16 µg/Kg/day i.v. continous infusion Results: median age of included patients was 59 years, with approximately 48 % male and 52% female, whithout significance in terms of recovery from FN (p = 1.00). 30% of the patients had prophilactic G-CSF administrated, but not significant for recovery from aplasia for included patients (p = 0.34). Median chemotherapy line responsible for FN was 2, and median cycle which produced FN was 3. Median level of neutrophiles (PMN) 450/mm3 and leucocytes (WBC) 1875/ mm3 at time of FN, 10 patients being in profound FN (PMN < 100/mm3). Median time to recovery was 25.5 hours for 72 included patients, with 1 patient dead whithout recovery. Predictive factors for shortened recovery were lower levels of reactive protein C (p < 0.001) and procalcitonine (p = 0.002) and higher WBC (p = 0.006) and PMN (p < 0.001) at time of administration of responsible chemotherapy regimen for FN. The earlier cycle responsible, between a line of chemotherapy, for FN, the better chance for patient to have shortened duration of FN (p < 0.0001). Conclusions: continous i.v. administration of G-CSF could represents a very effective alternative for patients with profound febrile neutropenia, with the shortest ever reported interval for neutrophiles recovery
Pharmacokinetic/pharmacodynamic assessment of a proposed biosimilar MSB11455 versus the currently licensed pegfilgrastim: A randomized, double-blind trial.
Authors: Lickliter Jason D. , Griffin Paul , Vincent Emmanuelle , Schuler Armin , Harrison-Moench Eleanor , Stahl Michael , El Bawab Samer , Kanceva Radmila , Gascón Pere
Published: May 2019

e14514

Background: MSB11455 is a proposed biosimilar to the currently licensed pegfilgrastim (Neulasta). This phase I study (NCT03251248) assessed the pharmacokinetic (PK)/pharmacodynamic (PD) bioequivalence of MSB11455 to Neulasta. Methods: Healthy volunteers were randomized to one of two crossover sequences, MSB11455/Neulasta or Neulasta/MSB11455. Subjects received a single subcutaneous dose of either MSB11455 or Neulasta (both 6 mg/0.6 mL) on Day 1 of each study period. Samples for PK/PD analysis were taken predose and up to Day 16 postdose. Immunogenicity samples were taken predose and up to Day 84 postdose. Safety was assessed throughout the study. Results: 244 subjects were randomized and received both treatments. For all primary PK/PD parameters 90% repeated confidence intervals of the geometric mean ratio of MSB11455 versus Neulasta were within the pre-defined equivalence range (80.00%–125.00%): AUC0–∞ (96.59, 112.82), AUC0–last (97.29, 113.96), Cmax (97.13, 114.99), Emax (98.74, 102.39) and AUE0–t (97.30, 100.23). Safety and tolerability as well as immunogenicity were comparable between treatment sequences. No filgrastim-specific neutralizing antibodies were detected in either treatment sequence. Conclusions: PK/PD equivalence of MSB11455 and pegfilgrastim was demonstrated with comparable immunogenicity, safety, and tolerability. This study supports the biosimilarity of MSB11455 to Neulasta. Clinical trial information: NCT03251248.
A highly efficient, low-cost, novel multicomponent nanosystem for rapid enumeration of circulating tumor cells.
Authors: Khandare Jayant , Qayyumi Burhanuddin , Prabhash Kumar , Chavan Rohit , Gourishankar Aland , Khatun Zamila , Mair Manish , Singh Arjun , Vasudevan Aravindan , Sagare Ajit , Chidara Vamshi , Bharde Atul , Chaturvedi Pankaj
Published: May 2019

e14516

Background: ‘Liquid biopsy’ technologies are unaffordable and unavailable in developing countries despite having highest cancer burden and mortality rates. Current Circulating Tumor Cell (CTCs) technologies sustain clinical concerns of a) non-specificity b) low efficiency c) high blood volume requirement d) long turn-around time, and d) exorbitant cost (~$900-1400). We report, an extremely low cost, innovative nanosystem for rapid enumeration of CTS with higher specificity and efficiency. Methods: We designed a nanosystem mediated by conjugation of anti-EpCAM through multi-reactive glutathione spacer, carbon allotrope and amine terminated dendrimer. The platform was evaluated for enhanced aqueous dispersibility and increased interaction with CTCs for rapid isolation and enumeration in 100 Head and Neck Cancer (HNC) patients having primary tumor sub-sites-oral cavity, larynx, hypopharynx, oropharynx, nasopharynx, salivary gland, and thyroid. The captured cells were immuno-stained and the optimal fluorescence acquisition intensity was validated in accounting CTCs with CK18 protein expression. There was complete elimination of the false positive normal cells (NC) count to CTCs by our method. The analysis was performed with only 1.5 ml of collected blood samples. Results: The CTC distribution in cohort study ranged from 1 - 85 cells per 1.5 mL of blood. In more than 80% of patient’s CTCs, the quantitative estimation of anti-CK18 protein over-expression indicated ~10-fold higher intensity over to NCs. As compared to treatment naive, recurrent, and disease-free patients, the spread of CTC number across the clinical range appeared to be tight (close to mean value). The CTC enumeration sensitivity linearity was ~99.2%, and the complete enumeration process time was under 03 hours/1.5 ml of blood. Consequently, efficient, rapid and yet affordable CTC platform was designed and clinically validated. Conclusions: ‘OncoDiscover’ liquid biopsy technology for CTC enumeration is poised to revolutionize the liquid biopsy due to its high sensitivity and affordability (~ $120) and shall resolve a major unmet medical need in impoverished world.
Systemic immune-inflammation index to predict prognosis of elderly patients with newly diagnosed solid tumors.
Authors: Li Chan , Tian Wei , Zhao Feng , Li Meng , Ye Qin , Wei Yuquan , Li Tao , Xie Ke
Published: May 2019

e14517

Background: Cancer in the elderly has become a common problem due in part to the increase in life expectancy. Compared to younger counterparts, the biological characteristics of tumors and their responsiveness to therapy may differ in elderly patients, and the elderly also can have a decreased tolerance to anticancer therapy. In addition, there is less evidence from clinical trials to guide physicians in treating aged patients with solid tumors. It would be highly useful to have a simple and powerful indicator of their prognosis. In this paper we evaluated the predictive value of the Systemic Immune-inflammation Index, SII, in determining the one-year survival in elderly patients with newly diagnosed solid tumors. Methods: Data from 118 patients, 70 to 89 years of age with newly diagnosed solid tumors in 2014 were retrospectively analyzed. The patients were divided into two groups according to age: (1) 70-75 years of age and (2) over 75 years of age. We calculated SII from the equation, SII = P x N/L, where P, N and L are the preoperative peripheral blood platelet, neutrophil and lymphocyte counts per liter respectively. The optimum cutoff point for SII for a favorable prognosis was determined to be 390×109 cells/L. For evaluation of SII as a prognostic indicator, the patients were divided into high SII ( > 390×109 cells /L) and low SII (≤ 390×109 cells/L) groups. Individual values were used to determine the relationship between SII and one-year survival in the two age groups. Results: A high SII > 390×109 cells/L was correlated with poor one-year survival (χ² = 7.658, P = 0.006). Patients with low SII had improved survival. Conclusions: SII was a robust indicator of one-year survival in elderly patients with newly diagnosed solid tumors. Patients in the high SII group showed poor prognosis compared to patients with a low SII score. [Table: see text]
EGF (epidermal growth factor) receptor expression in blood: Prognostic biomarker in overall survival (OS) of patients with non-metastatic triple negative (TN) breast carcinoma.
Authors: De Araujo Rogerio Agenor , da Luz Felipe Andrés Cordero , do Amaral Pedro Augusto , Marinho Eduarda da Costa , Nascimento Camila Piqui , Mosca Etelvina Rocha , Delfino Patricia Ferreira Ribeiro , Antonioli Rafael Mathias , Mendes Thais Rezende , Araújo Breno Jeha , Silva Marcelo José Barbosa , Neto Morun Bernardino
Published: May 2019

e14518

Background: EGF acts on tumor progression and its receptor can be measured by immunohistochemistry, or in the blood by immunoenzymatic method. This study quantified the expression of blood EGF receptors (bEGFR) in the molecular subtypes of breast carcinoma and their impact on OS. Methods: Prospective study with 125 patients (pts) and 89 healthy women (controls). All of them signed the free and detailed consent form. The blood collection was performed before the surgery (stages I–10pts; II–59pts) and before neoadjuvant chemotherapy (stage III-41pts) or palliative (stage IV–15pts). All the patients have made the protocol of a public service treatment. Measurement of bEGFR expression was done by the ELISA method. The statistical significance was set at p < 0.05. Results: In a follow-up of 29.9 months (8-44m) 23 deaths occurred at 15.8 months (8-34.5m); 58pt had bEGFR < 40ng/mL (E40-) and 67pt ≥40ng/mL (E40 +). TNs (28pt) had lower OS than Luminal (A = 29pts; B = 44pts) and HER2 (24pts) (p = 0.0020). But in TN E40- the significant difference in OS disappeared (TN versus Luminal A: p = 0.0785; TN versus Luminal B: p = 0.1995; TN versus HER2: p = 0.3017); which did not occur in TN E40 + (TN versus Luminal A: p = 0.0009; TN versus Luminal B: p = 0.0241; TN versus HER2: p = 0.0014). In OS of non-metastatic 24 TN (3pts stages 0/I; 13pts II; 8pts III) there was significant difference according to bEGFR expression (E40+ with lower OS than E40-, HR:5.49; 95%CI:1.26–15.07;p = 0.0159). In binary logistic regression, bEGFR was considered to be a significant predictor of death in TN non-metastatic, p = 0.008 (bEGFR of 30ng/mL = death probability of 6.7% until bEGFR of 80ng/mL = probability of death of 99.8%). There were no significant differences in bEGFR between patients and controls, among grades of histological differentiation, or in those with ki67 > 14%. There was no difference in OS according to bEGFR in stages I, III or IV, except in stage II (E40+ with lower OS than E40-, HR:4.248;95% CI:1.229–14.68; p = 0.0460). Conclusions: Increased expression of bEGFR in non-metastatic TNs is a predictor of lower OS, with statistical significance.
Association of forkhead box protein O1 and paired box gene 3 overexpression with prognosis in patients with cervical cancer.
Authors: Cho Hanbyoul , Han Gwan Hee , Chay Doo Byung , Kim Jaehoon
Published: May 2019

e14519

Background: Transcriptional factor FOXO1and PAX3 has been reported to play an imported role in human cancer, but the role in cervical cancer has not yet been clarified. Here, we evaluated the functional role of FOXO1 with cervical cancer cells and the expression together with clinical significance of FOXO1 and PAX3 among cervical lesions was investigated. Methods: In vitro assessment of cell functions by cell viably assay, cell migration and invasion assay was evaluated using FOXO1 knockdown cervical cancer cell lines. Immuno-histochemical (IHC) staining analyses of FOXO1 and PAX3 were performed with tissue microarray (TMA) consisting of 209 cervical cancers, 366 high grade cervical intraepithelial neoplasias (CIN), 130 low grade CIN and 328 matched nonadjacent normal cervical epithelial tissues. The clinical significance was investigated by comparing the data with various clinicopathologic characteristics, including survival in cervical cancer. Results: In vitro result revealed that knockdown of FOXO1 was associated decreased cell viability ( p< 0.001), migration ( p< 0.001) and invasion ( p< 0.05) supporting the oncogenic role of FOXO1 in cervical cancer. FOXO1 and PAX3 expression was significantly higher in CIN (both p< 0.001) and cancer tissue (both p< 0.001) than in normal tissue. FOXO1 expression showed significant correlation with PAX3 (Spearman’s rho = 0.377, p< 0.001) in a cervical cancer. Multivariate analysis indicates that FOXO1 expression (hazard ratio = 4.01 [95% CI, 1.22–13.10], p= 0.021) and advanced FIGO stage (hazard ratio = 3.89 [95% CI, 1.35–11.19], p= 0.012) were independent prognostic factor on overall survival. Conclusions: This study reveals the association between FOXO1 and PAX3 expression with clinicopathologic variables, including survival of cervical cancer patients. Our results not only suggest the promising potential of FOXO1 as a prognostic and survival marker, but also warrant further studies on a possible link between the biological function of FOXO1 and the pathogenesis of cervical cancer.
Detection of low abundant mutants from circulating tumor DNA of plasma, pleural effusion, and cerebrospinal fluid of lung cancer patients using a novel mutant-capture enrichment approach.
Authors: Lin Rui , Pu Yue , Mao Li
Published: May 2019

e14520

Background: In the era of precision medicine, liquid biopsy analysis is well accepted based on advantages including availability, non-invasiveness, and non-heterogeneity. However, the circulating tumor DNA (ctDNA) in liquid biopsy is diluted by a large excess of wild-type alleles, which necessitates high sensitivity approach for ctDNA detection. In addition, ctDNA analysis from different liquid biopsy samples need to be evaluated. Methods: We have developed a novel mutant-capture based method, termed PErsonalized Analysis of Cancer (PEAC), for high sensitivity detection of cancer driver mutants at abundance as low as 0.01-0.1% for circulating free DNA (cfDNA) standards. ctDNA samples were extracted from body fluids of lung cancer patients including plasma, pleural effusion and cerebrospinal fluid. EGFR mutants predictive of EGFR tyrosine kinase activity were enriched using PEAC technology, and analyzed using Sanger sequencing. Results: Plasma ctDNA samples B7110003, B7110010, and B7112012 had no or barely detectable L858R mutation, which was enriched to 50-90% after PEAC and readily detected by Sanger. T790M was undetectable before PEAC in plasma sample B7112052 and became 50% after PEAC enrichment. Pleural effusion samples E8106029 and E8111305 had dominated L858R and T790M peaks, respectively, in Sanger chromatograms after PEAC, which was almost to the background levels prior to PEAC. Interestingly, both EGFR L858R and T790M mutants were detected in pleural effusion sample E8106029 after PEAC; the sample was from a patient who had previously treated with an EGFR tyrosine kinase inhibitor (TKI), suggestive of resistance developed after target therapy and the utility of PEAC in monitoring patient’s response to EGFR TKI. In addition to enriching point mutations, we also established enrichment of the most frequently occurred EGFR 19 deletion, E746_A750del (c. 2235_2249 del15), which were dominant after PEAC enrichment of ctDNA from plasma samples (B8101186 and B8101241), pleural effusion (E8108088) and cerebrospinal fluid (C8108095); the mutants were undetectable without PEAC enrichment. Conclusions: PEAC technology can enrich ctDNA from body fluids in lung cancer patients and allow detection of low abundant mutants predictive for EGFR TKI therapy. With further validation, the technology may improve current detection methods used in clinical practice.
Elevated plasma cell-free DNA copy number variations to predict survival for cancer patient with spinal metastasis.
Authors: Chen Su , Yang Minglei , Zhong Nanzhe , Ziliang Qian , Jiao Jian , Xiao Yasong , Jiang Dongjie , Zhou Zhenhua , Wang Tianzhen , Lou Yan , Xu Wei , Wan Wei , Wu Zhipeng , Yang Cheng , Wei Wei , Liu Tielong , Zhao Jian , Wei Haifeng , Yang Xinghai , Xiao Jianru
Published: May 2019

e14522

Background: Approximately 70% of patients with cancer have evidence of spinal metastatic disease at the time of their deaths. The spinal column is the most common location among osseous sites for metastatic deposits. Expected overall survival is the major determinant of treatments for those ones suffering from intractable pain, neurologic deficits, and even paralysis. We sought to evaluate copy number variations (CNV) in spinal metastatic cancer via cfDNA and determine if CNV is associated with prognosis and treatment decision. Methods: In this study, 314 patients with pathologically confirmed spinal metastatic cancers were recruited since November 2015. 314 plasma samples were sent to low-coverage genome-wide sequencing of cfDNA from plasma followed by a customized bioinformatics workflow UCAD. Tokuhashi score was also evaluated for each patient before surgery. Statistical correlation with clinical index like prognosis was estimated. Results: 280 evaluable data were collected (34 samples failed Quality Control). The median follow-up time is 276 days. 114 (40.7%) patients died during follow up till December the 25th, 2018. Elevated CNVs was found in 109 (38.9%) patients, including 9(69.2%) head&neck, 15(46.9%) liver and gallbladder, 27(44.3%) lung, 7(38.9%) breast, 2(28.6%) prostate, 5(19.2%) thyroid cancer, 4 (10.5%) kidney and 20(40.0%) cancer with unknown primary site. Further analyses showed that patients with elevated CNVs were found with worse survival. The median overall survival (OS) was 298 (95% CI: 258-422) days, as compared with those low-CNVs status with median OS 657 (95% CI:433-NA) days (Hazard ratio = 3.73 [95% CI: 2.22-6.27], P < 0.01). Especially for patients with low Tokuhashi score (≤8) who have the predictive survival less than 6 months, CNVs score distinguish those well-prognosis patients with median OS 433 (95% CI: 308-NA) days from the worse survival group with median OS 285(95% CI:243-348) days (Hazard ratio = 2.42 [95% CI:1.38-4.25], P = 0.013). Conclusions: We present the largest cfDNA genomic characterization of spinal metastatic cancers. Specific CNVs features are enriched in spinal metastasis cancers with different primary sites. Elevated CNVs in plasma cfDNA is significantly associated with worse survival in a large spinal metastatic cancer cohort. This demonstrates that cfDNA CNVs could be a useful marker in estimating the survival time of spinal metastasis cancer patients for whom the outcome is mainly dependent on the selection of proper treatment.
Epigenomic profiling of plasma cell-free DNA (cfDNA) to predict bladder cancer recurrence after radical cystectomy.
Authors: Lu Yi-Tsung , Gooden Gerald C , Tew Ben Yi , Buckley David , Siegmund Kimberly , Daneshmand Siamak , Salhia Bodour , Goldkorn Amir
Published: May 2019

e14525

Background: Muscle invasive bladder cancer (MIBC) frequently recurs despite radical cystectomy, and new biomarkers are needed to guide treatment. DNA methylation plays a key regulatory role in cancer initiation and progression and has been correlated with prognosis in multiple cancer types. We hypothesized that methylation patterns in cfDNA can predict recurrence after curative cystectomy. Methods: Using a clinically annotated USC bladder cancer tissue archive under an IRB approved protocol, we selected 40 patients whose cancer relapsed within 3 years of radical cystectomy, as well as 46 age-matched and stage-matched patients who did not recur after 3 years of follow-up. cfDNA was extracted from the archival plasma, and samples were pooled for whole genome bisulfite sequencing (WGBS) based on pathologic stage and recurrence status. Differentially methylated regions (DMRs) was identified between recurrence and non-recurrence groups in each stage. Results: Pooled analysis across stages yielded 729 DMRs, of which 487 (67%) were hypomethylated in the pooled recurrence group. Of the 729 DMRs from the pooled analysis, 63.5% were in gene bodies, 17.1% were in promoter regions and 47.9% were at CPG islands. Unsupervised clustering of these DMRs accurately distinguished the recurrence groups. Stage-specific analysis yielded DMR sets ranging from 6412 to 9532 DMRs per stage, with a similar trend of global hypomethylation in the recurrence groups. Basespace Correlation Engine analysis revealed that methylation in our recurrence groups (pooled and by stage) was correlated with bladder cancer methylation in publicly available data and also correlated with expression in the TCGA bladder cancer dataset. Conclusions: WGBS was successfully performed in MIBC using cfDNA extracted from clinically annotated archival plasma. Methylation patterns were correlated with publicly available methylation and expression datasets, and cfDNA hypomethylation pre-cystectomy in our cohort was associated with cancer recurrence. A subset of maximally discriminatory DMRs has been selected to define a methylation signature that is currently undergoing validation as a biomarker predictive of recurrence in MIBC.
Circulating tumor DNA molecular profiling in rare cancer patients from the MASTER KEY Project.
Authors: Okuma Hitomi Sumiyoshi , Yonemori Kan , Seo Takuji , Noguchi Emi , Wakakuwa Keiko , Kato Ken , Ichikawa Hitoshi , Kohno Takashi , Hirakawa Akihiro , Kuchiba Aya , Shibata Taro , Sukigara Tamie , Nakamura Kenichi , Mano Hiroyuki , Nishida Toshirou , Fujiwara Yasuhiro
Published: May 2019

e14532

Background: In April 2017, MASTER KEY Project, composed of a prospective registry study part and a multiple clinical trials part, was established to promote treatment development for rare cancers, which is lacking standard or investigational therapeutic options. Circulating tumor DNA (ctDNA) analysis by next-generation sequencing (NGS) has provided new insight into personalized medicine in a more accessible, non-invasive manner; however, most reports are focused on common cancers. We report genetic alterations detected by ctDNA NGS analysis in rare solid cancers. Methods: Prospectively consented patients (pts), also enrolled in MASTER KEY registry study, had ctDNA NGS testing at a CLIA-certified lab (Guardant360) for point mutations, indels, copy number amplifications, fusions, and microsatellite instability. Alterations were assessed for incidence according to cancer type, functional impact, therapeutic implications, and comparison with tissue NGS. Main inclusion criteria: 1) age ≥16, 2) histological diagnosis of rare cancer (annual incidence less than 6 cases per 100,000 population), cancer of unknown primary (CUP), or rare tissue subtypes of common cancers, 3) active metastatic / unresectable cancer, 4) a written consent. Results: From Nov. 2018 to Jan 2019, 50 pts had Guardant360 testing. Diseases included: soft tissue sarcoma (28), ovarian carc. (7), CUP (4), salivary gland tumor (3), thyroid carc. (2), GIST (1), adrenal cortical carc. (1), rare subtype of GI tract (1), malignant mesothelioma (1), nephroblastoma (1), NET (1), NUT carc. (1). All Japanese, male/female = 14/36, median age 61, ECOG performance status 0/1/2/3 = 30/19/0/1. 76% of pts (38/50) had ≥1 alteration detected, with median number of 2 alterations (range: 0-9), with VAF(0.1-60.3%); 87%(33/38) of those pts had a variant most likely pathogenic; 61% (20/33) of those pts had a variant potential for clinical action. Mean TAT was 10.3 days. 19 pts had tissue NGS testing and in 3 pts, alterations were detected by ctDNA NGS but not by tissue NGS. Updated results of 100 patients will be presented at the conference. Conclusions: Most rare cancer patients with advanced disease had a detectable genomic alteration by Guardant360. Clinical trial information: UMIN000034394.
Cell-free DNA methylation markers for noninvasive early detection of nasopharyngeal carcinoma.
Authors: Roy Dhruvajyoti , Taggart David J , Zheng Lianghong , Liu Dan , Li Gen , Li Mingzhen , Zhang Kang , Van Etten Richard A.
Published: May 2019

e14537

Background: Nasopharyngeal carcinoma (NPC) is one of the most prevalent malignancies among populations native to Southeast Asia, the Mediterranean Basin and the Arctic. Early diagnosis of NPC is predicted to improve survival. The identification of cancer-specific DNA methylation patterns of cell-free DNA (cfDNA) isolated from blood samples is an established approach for detecting various cancers. In the present study, we evaluated the performance characteristics of a previously identified NPC methylation marker panel for the diagnosis of nasopharyngeal carcinoma. Methods: Retrospective samples were obtained for 168 subjects, including: 59 subjects diagnosed with NPC (Stage I to IV), 14 subjects diagnosed with benign nasopharyngeal disease and 43 healthy subjects. In addition, sample were obtained for 52 subjects diagnosed with breast, colorectal, liver or lung cancer. Samples were provided to the laboratory blinded for DNA methylation analysis by using the IvyGene Platform. Results: A total of 57 of the 59 samples drawn from subjects with NPC were correctly identified for an overall sensitivity of 97%, with little difference between the sensitivity of detecting Stage I to Stage IV nasopharyngeal carcinoma (range 92% to 100%). For subjects diagnosed with other cancers, 85% of colorectal cancer samples, 82% of lung cancer samples, 93% of both breast cancer and liver cancer samples, were correctly identified as negative for NPC, for a total calculated analytical specificity of 86%. Additionally, all 43 samples drawn from healthy donors and 14 samples drawn from subjects diagnosed with benign nasopharyngeal disease were correctly identified as negative for nasopharyngeal carcinoma for a combined specificity of 100%. Conclusions: The NPC methylation panel was demonstrated to be both sensitive and specific for the detection of nasopharyngeal carcinoma. The potential of cfDNA methylation markers for the early detection of nasopharyngeal carcinoma is predicted to improve patient outcomes through earlier detection of the disease.