bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of germline cancer predisposition variants during clinical ctDNA testing.
Authors: Stout Leigh Anne Anne , Kassem Nawal , Hunter Cynthia , Philips Santosh , Radovich Milan , Schneider Bryan P.
Published: May 2020

e15555

Background: Next-generation sequencing (NGS) of circulating tumor DNA (ctDNA) is a non-invasive method to guide therapy selection for cancer patients. Identification of inherited germline cancer predisposition mutations that have significant implications for at-risk relatives may be missed during routine ctDNA testing. Allele frequency has the potential to enhance the likelihood that a mutation is germline; and is often reported in many NGS tests from ctDNA. Here, we report on the fidelity of allele frequency in ctDNA as a predictor for pathogenic germline variant carriage. Methods: ctDNA sequencing of patients with metastatic cancer from the Indiana University Health Precision Genomics Program was performed using the FoundationOne Liquid assay. All variants detected by the ctDNA assay report were considered. All patients also had germline testing information and pathogenicity of germline variants were determined using ClinVar. Germline variants with conflicting interpretations were manually reviewed to determine pathogenicity. Comparisons between ctDNA results with known germline status were performed. Results: Of 91 previously identified germline cancer predisposition variants, 36 (40%) were also identified by ctDNA analysis. All germline variants that were tested for in the ctDNA assay (n = 36, 100%) were identified. When detected, the allele frequencies of detected germline variants in the ctDNA ranged from 39-87.6% with an average of 52.1%. Conversely, 111 of 160 (69%) variants identified by ctDNA analysis with allele frequency between 40-60% in a cancer predisposition gene were found to be germline in origin (regardless of pathogenicity). Variants in the BRCA2, BRCA1, and CDH1 genes were most likely to be germline in origin (26/27 [96%], 20/22 [91%], 13/15 [87%], respectively). Variants in the TP53 and APC genes were least likely to be germline in origin (9/36 [25%] and 1/6 [17%], respectively). There was an 85% (95/111) concordance in actionability between the somatic testing lab and ClinVar germline classifications. Of the 16 discordant variants, 100% were determined to be actionable by the somatic testing lab but not actionable in ClinVar. Conclusions: ctDNA allele frequency can alter the likelihood that a variant is germline. Importantly, however, this testing is far from comprehensive and should not be used as a replacement for germline testing. Variants with allele frequency between 40-60% in cancer predisposition genes should induce a high level of suspicion for germline status.
Detection of residual disease and recurrence in early-stage non-small cell lung cancer (NSCLC) patients using sensitive personalized ctDNA sequencing assays.
Authors: Heider Katrin , Gale Davina Gillian , Marsico Giovanni , Ruiz-Valdepeñas Andrea , Sharma Garima , Perry Malcolm , Osborne Robert , Howarth Karen , Rundell Viona , Belic Jelena , Wulff Jerome , Harden Susan , Rassl Doris M , Rintoul Robert , Rosenfeld Nitzan
Published: May 2020

e15560

Background: Detection of residual circulating tumour DNA (ctDNA) in patient plasma following curative intervention for localized non-small cell lung cancer (NSCLC) could identify patients who are at higher risk of relapse. These patients may benefit from adjuvant treatment, even if they have no macroscopic disease identified by radiographic imaging, which is the current standard of care. Here we evaluate the performance of the Inivata personalized sequencing assays to detect ctDNA in a cohort of 90 patients with early-stage NSCLC undergoing treatment with curative intent. Methods: The Inivata assay uses a highly sensitive next-generation sequencing platform, to identify tumor-specific variants from exome sequencing of tumor tissue and to track up to 48 patient-specific mutations in plasma specimens by multiplex PCR and ultra-high-depth next-generation sequencing. Samples from 90 patients with Stage I-III NSCLC who underwent radical treatment with curative intent, either surgery or radiotherapy ± chemotherapy, were collected as part of the LUng cancer - CIrculating tumor DNA (LUCID) study. Results: 350 plasma samples from 90 patients were analyzed using the Inivata assay, including samples collected before and after treatment and at subsequent follow-up visits. ctDNA was detected in pre-treatment samples in 38% of 32 patients (12/32) with Stage I NSCLC and in 90% of 21 patients (19/21) with Stage II/III disease, at allele fractions ranging from 6 parts per million (ppm, equivalent to 0.0006%) to over 20,000 ppm (equivalent to 2%). In plasma samples collected post-treatment, ctDNA was detected in close to 50% of cases. Conclusions: These findings highlight the Inivata assay is a sensitive method for detection of residual ctDNA and recurrence in early stage NSCLC. Initial detection rates ranged from 38% in Stage I disease to 90% for patients with Stage II/III disease prior to treatment, including detection of ctDNA to levels as low as a few parts per million. ctDNA was detected in at least one post-treatment timepoint in close to 50% patients. Together with additional data to be presented from the full 90 patient cohort, this suggests a possible route to improving treatment and designs of adjuvant trials for early stage NSCLC by detection of residual disease post-treatment and monitoring for early detection of relapse.
A phase I study of the novel immunotoxin, MT-5111, in subjects (subj) with HER-2 positive tumors: Interim results.
Authors: Wainberg Zev A. , Barve Minal A. , Hamilton Erika Paige , Brenner Andrew Jacob , Valdes Frances , Mita Monica M. , González Valeria , Anand Banmeet S. , Burnett Christine , Pelham Joshua , Waltzman Roger J. , Strack Thomas , Van Tine Brian A.
Published: May 2020

e15567

Background: Engineered toxin bodies (ETBs), composed of a Shiga-like Toxin A subunit genetically fused to antibody-like binding domains, can force receptor internalization, self-route through intracellular compartments to the cytosol, and induce potent cell-kill via the enzymatic and permanent inactivation of ribosomes. MT-5111, a de-immunized 55 kD ETB targeting HER2 in solid tumors, binds to an epitope distinct from and non-competitive with trastuzumab and pertuzumab. Methods: The primary objective is to determine the maximum tolerated dose (MTD) of MT-5111 monotherapy in adults with advanced HER2+ solid tumors. Secondary objectives are pharmacokinetics, efficacy, and immunogenicity. Part 1 will identify the MTD via MT-5111 dose escalation (0.5, 1.0, 2.0, 3.0, 4.5, 6.75, 10 µg/kg/dose) according to a modified 3+3 design. Part 2 will further evaluate the safety of MT-5111 at the recommended phase 2 dose (RP2D) in 3 separate cohorts of subjs with a HER2 immunohistochemistry score ≥2 in a metastatic lesion of primary breast, gastroesophageal, or other cancers. All subjs receive MT-5111 as a 30-minute IV infusion on Days 1, 8, and 15 of each 21-day treatment (tx) cycle (C) until disease progression, unacceptable toxicity, death, or withdrawn consent. Details can be found on clinicaltrials.gov (NCT04029922). Results: The first cohort (0.5 µg/kg/dose) enrolled 4 subjs (metastatic breast cancer, n = 2; metastatic cholangiocarcinoma, n = 2). Three subjs were female and the mean age was 69 years (median 65, range 64-78). Subjs received a mean of 5 prior lines of therapy (median 4.5, range 3-8). Three subjs completed C1 of tx without dose-limiting toxicities; 1 subj was inevaluable. Two subjs had progressive disease in C2. A total of 23 AEs occurred in 4 subjs; all were grade (G) 1-2 except one G3 event of hypertension in a subj with a history of hypertension. There were 2 tx-related AEs (G1 chills; G2 aspartate aminotransferase increased in the setting of progressive liver metastases). There was 1 serious, non-tx-related AE (G2 dyspnea) that occurred in the inevaluable subj. No cardiac AEs were noted, nor clinically significant changes in cardiac biomarkers, an important safety parameter given non-human primate toxicity. Conclusions: MT-5111 appears to be well tolerated at the lowest dose with no apparent cardiotoxicity to date. Drug concentrations are expected to be below the level required for in vitro tumor cell killing. Safety and efficacy data from subsequent dose escalation cohorts are expected by May 2020. Clinical trial information: NCT04029922.
Validation of an exome and transcriptome based diagnostic platform enabling clinical cancer therapy selection and emerging composite biomarkers for immunotherapy.
Authors: Saldivar Juan-Sebstian , Harris Jason , Desai Sejal , Ayash Erin , Tandon Prateek , Marfatia Twinkal , Power Robert , Morra Massimo , Chinnappa Manju , Clark Michael James , McClory Rena , Chen Richard
Published: May 2020

e15583

Background: While immunotherapy has become a pillar of cancer treatment, diagnostic biomarkers that consistently predict patient response to these therapies have remained elusive. There is an increasing need for the development of integrative, composite biomarkers that can model the complex biology driving response and/or resistance to immunotherapy more effectively than existing single-analyte approaches. However, the majority of current cancer diagnostic panels, with their focus on a small set of genes, provide limited ability to support these emerging advanced biomarkers. Methods: To address these limitations, we developed and validated NeXT Dx, a comprehensive enhanced exome and transcriptome based diagnostic platform designed to simultaneously characterize tumor and immune genomics from a single limited FFPE sample. To achieve higher accuracy and sensitivity for an exome scale diagnostic platform, we developed an augmented exome assay that improves uniformity of coverage across all ~20,000 genes, including boosted coverage of 248 clinically-relevant cancer genes. We validated this assay using genomic DNA and RNA extracted from tumor-derived cell-lines, constructs, clinical FFPE samples, and proficiency testing samples. The assay utilizes > = 25ng of co-extracted DNA and RNA which were sequenced using Illumina NovaSeq instruments at our CAP-accredited, CLIA-certified laboratory. Additional assay enhancements for HLA, immune repertoire, and oncoviruses were designed to further optimize the platform for immunotherapy biomarker discovery applications. Results: Validation of NeXT Dx demonstrated a performance of 99.5% sensitivity and 99.8% positive predictive value (PPV) for SNVs with > = 5% AF; 98.7% sensitivity and 97.4% PPV for indels with > = 10% AF; 97.2% sensitivity and 94.6% PPV for CNAs in samples with > = 30% tumor content; 94.9% sensitivity and 94.9% PPV for fusions; and a 2.1% error rate for MSI classification. TMB was calculated using gold-standard whole exome data from SNVs and indels. Typical median coverage depth was > 1,000X for 248 clinically-relevant genes, ~300X for the remaining (whole exome) footprint. Conclusions: With NeXT Dx, we demonstrate a exome/transcriptome scale diagnostic platform that can detect current clinical biomarkers with high sensitivity as well as support emerging, advanced biomarkers that integrate across both tumor and immune features.
Next-generation sequencing for genetic analysis of endobronchial ultrasound transbronchial needle aspiration samples in advanced lung cancer.
Authors: Min Xianjun , Li Mingwei , Liu Qiang , Yang Yingshun , Pei Guotian , Wang Shuai , Wang Xiaoyu , Wang Huina , Lou Feng , Cao Shanbo , Liu Jun , Huang Yuqing
Published: May 2020

e15586

Background: Next-generation sequencing (NGS) typically requires greater quantities of DNA than traditional molecular testing. Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration (EBUS-TBNA) is a minimally invasive technique with high sensitivity in the mediastinal staging of lung cancer. This study aimed to evaluate the adequacy of EBUS-TBNA in providing adequate size specimens for genetic mutations and immunotherapy biomarkers analysis in patients with lung cancer. Methods: Tissue samples from patients with advanced lung cancer were collected by EBUS-TBNA and were formalin-fixed paraffin-embedded. NGS assay was carried on with acornmed panel including 808 genes. PD-L1 expression through immunohistochemistry was assessed. Results: A total of 98 patients was enrolled, of which 74 (76%) were adenocarcinoma and 20 (20%) were squamous cell carcinomas. Among the patients, 108 samples (including multipoint puncture of different location and retest of the same patient) were obtained. NGS assay was completed successfully on 106 of the 108 samples (98.14%), and 97.96% of patients had successful testing, identifying an average of 11.3 mutations. With a multi-gene panel comprising up to 808 tumor related genes, actionable variations were found in 93 (86%) samples. Of these, the positive rates of actionable alterations in lung adenocarcinoma samples and lung squamous cell carcinoma samples were 90% and 79%, respectively. The tumor mutation burden (TMB), Microsatellite instability (MSI) and PD-L1 expression were found High in 20 (41%), 1 (2%), and 37 (72%) patients respectively. Conclusions: NGS assay can be successfully conducted with tissue samples obtained from EBUS-TBNA. NGS assay provides more comprehensive information on genetic mutations in tumors, which greatly assists therapeutic decision making for advanced lung cancer.
The dark age of single organ screening is over: CD24 is a novel universal simple blood test for early detection of cancer.
Authors: Shapira Shiran , Kazanov Dina , Ben Shimon Marina , Hay Levy Mori , Mdah Fatin , Asido Shir , Carmel Narin , Yossepowitch Ofer , Grisaru Dan , Fliss Dan , Isakov Ofer , Lahat Guy , Nachmany Ido , Gluck Nathan , Peer Michael , Wolf Ido , Arber Nadir
Published: May 2020

e15591

Background: More than 1.8 million new cancer cases are expected to be diagnosed in 2020. Only in the U.S, about 606,520 Americans are expected to die this year, which translates to about 1,660 deaths per day. However, when found early, in the asymptomatic phase, cancer is often effectively treated or even cured. The few cancer screens available today, including mammograms and colonoscopies, each look for a single type of cancer. CD24 is hardly expressed in normal cells (but to B-lymphocytes, differentiating neuroblasts and neutrophils) but is overexpressed in numerous human cancers. Aim: To establish a universal cancer screening blood test. Methods: Blood samples were obtained from consecutive patients and healthy volunteers. 1x106 leukocytes were stained using anti-CD11b-PerCp-Cy5.5 and anti-CD24-FITC and analyzed by flow cytometry (CyFlow Cube 6, Sysmex, Germany). Percentage of positive cells was determined by subtracting the percentage of CD24 and CD11b-positive cells (dual stain) from CD24-positive cells (single stain). The cut off for cancer detection was 25. Healthy subjects underwent a thorough evaluation at the health promotion center and integrated cancer prevention center. All cancers were verified histologically. Results: CD24/CD11b expression in the healthy population was evaluated in 337 women and 408 men, ages 20-85. There were 222 cancer patients. After excluding thyroid and bladder cancers, the assay detected 146 out of 196 (75%) cancers at all stages and even in 31 out of colonic adenomas (table). The levels of CD24 drop back to near normal level following successful surgery/chemotherapy. The test score was not affected by age, gender, fasting and the time of the day that the blood sample is taken. In 55 cases the test was performed twice, with similar results, as well as when the test was performed in another flow cytometry, after 1d, 1w, 1m after freezing . Conclusions: 1. CD24 is a universal cancer screening. 2. It can detect pre malignant lesions like adenomas in the colon. 3. Preliminary results suggest that it can serve as a predictive marker. [Table: see text]
Effect of UV-curable and injectable dendrimer matrix on tumor pharmacokinetics and survival in a hepatocellular carcinoma mouse model using interstitial photodynamic therapy.
Authors: Vuoti Sauli , Narasimha Kumar , Reinikainen Kai
Published: May 2020

e15598

Background: Intratumoral drug delivery in cancer treatment has shown to increase drug accumulation in tumors and decrease adverse effects of systemic therapies. The active agent is typically required to stay in contact with the tumor and thus attachment directly into the tumor or to the surrounding tissue is warranted. In addition, a long-acting release of the active drug is vital. Dendrimers are nano-sized, radially symmetric molecules with homogeneous and monodisperse structures, which can used to encapsulate drug molecules for controlled release with the aid of external stimuli at physiological pH. Methods: We prepared a dendrimeric drug delivery matrix based on a PEG-core. The model drug (5-fluorouracil or doxorubicin) was covalently bonded to the core. The release of the drug was observed up to 29 days in vitro. The outer shell of the matrix was partially modified to obtain a fast UV-curing and subsequent attachment to the tumor tissue utilizing interstitial photodynamic therapy. We compared the pharmacokinetics, tissue distribution and antitumor efficacy of conventional systemic therapy and the intratumoral therapy in xenograft-bearing mice. In addition, we monitored blood parameters and body weight of the animals for adverse effects. Results: The dendrimer matrix caused significantly greater inhibition of human hepatocellular carcinoma xenograft tumor growth when compared with systemic therapy, without increasing toxicity. The matrix enabled a significant increase in drug accumulation in tumors, and markedly extended the survival of mice compared with systemic treatments. A larger decrease in blood parameters as well as spleen weight was observed in the systemic therapy group especially when using 5-fluorouracil. Conclusions: Our study shows that the intratumoral treatment strategy based on a novel UV-curable dendrimer hydrogel matrix enhances antitumor activity by improving pharmacokinetics and drug accumulation and increases efficacy in comparison to conventional systemic therapy. These results highlight the potential use of dendrimer hydrogels in the treatment of hepatocellular carcinoma.
LCN2 and colon cancer — Have we hit the jackpot.
Authors: Srinivasalu Vijay Kumar , Chaudhary Nazia , . Bhagyashree , Dwivedi Nehanjali , P K Smitha , Thorat Rahul , Batulla Christopher , Dhar Sujan Kumar , Dalal Sorab N , Das Manjula
Published: May 2020

e15608

Background: Lipocalin2 (LCN2, also known as neutrophil gelatinase-associated lipocalin) is a protein that in humans is encoded by the LCN2 gene. Its abnormal expression serves critical roles in EMT transition, angiogenesis, cell migration and invasion in many cancers. We aim to assess the in vitro and in vivo effects of LCN2 as a potential chemo and radiosensitizer. Methods: Normalized RNAseq RSEM values of LCN2 were compared between normal and tumour samples from TCGA. Differences between median expression levels were assessed using Wilcoxon rank sum test. Kaplan-Meier model was used for survival analysis. Immune cell population in publicly available Colon Adenocarcinoma dataset was estimated using MCP Counter tool. Cell systems used to experimentally study the role of LCN2 in therapy resistance and tumor progression were HCT116, HT29 and DLD1. PKP3 and/or LCN2 were knocked down by shRNA. Tumor regression and therapy (5FU and radiation) sensitivity upon Anti-LCN2 treatment were demonstrated in Xenograft mouse models. Results: Analysis of 23 TCGA datasets containing gene expression data for both tumour and adjacent normal samples indicated that LCN2 levels are elevated in colon tumors. Colon cancer cell line HCT116 derived PKP3 knock-down or LCN2 over-expressing cells showed therapy resistance. A comparison of the tumor cell lines HCT116, HT29 and DLD1 show that increased LCN2 expression correlates with therapy resistance. LCN2 levels correlated with resistance to 5FU (p = 0.006) and its ability to clear ROS (p < 0.05) in vitro. Inhibiting LCN2 led to a decrease in invasion in vitro (p = 0.0005), increased sensitivity to 5FU in vitro (p = 0.001) and inhibition in tumor growth and increased sensitivity to 5FU and radiation (p = 0.005) in xenograft mouse models. On MCP counter analysis of TCGA, in Colon adenoca the normal samples show a correlation between LCN2 expression and T-cells (Pearson r = 0.45, p = 0.0028) and with the T-cell chemoattractant CXCL10 (Pearson r = 0.5, p < 0.0001). Such correlations are broken in tumour samples. Conclusions: LCN2 expression leads to chemo and radio resistance in colon cancer cell lines and xenograft mouse models. Inhibiting LCN2 function can inhibit tumor progression and sensitizes tumors to radiation and 5FU. These results suggest that LCN2 expression could be a marker that can be used to determine the choice of therapy offered to patients and that LCN2 could serve as a therapeutic target that sensitizes cells to radio and chemotherapy. LCN2 affects tumor progression and therapy sensitivity probably through T cell mediated immune pathway.
Prevalence of iron deficiency in cancer patients: The French prospective CARENFER study.
Authors: Luporsi Elisabeth , Carnot Aurelien , Massard Vincent , Morin Sophie , Chauffert Bruno , Turpin Anthony , Trochu Jean-Noël , Cacoub Patrice
Published: May 2020

e15617

Background: Despite the potential deleterious consequences of iron deficiency (ID) in patients with cancer, under-diagnosis is frequent and existence of ID based on both serum ferritin concentration and transferrin coefficient saturation (TSAT) index is rarely determined. The CARENFER Study aimed to assess prospectively the prevalence of ID, anemia and iron deficiency anemia (IDA) in cancer patients. Methods: The study was conducted in France from May to July 2019, as a prospective cross-sectional survey carried out in 15 oncology units. The analysis focused on 1221 patients with different types of solid malignant tumors. Serum ferritin and iron concentrations, TSAT index and hemoglobin (Hb) level were determined. Based on ESMO 2018 Guidelines, ID was defined as ferritin < 100 μg/L or TSAT < 20%, and then categorized as either absolute ID (ferritin < 100 μg/L) or functional ID (ferritin ≥ 100μg/L and TSAT < 20%). Anemia was defined as Hb level ≤ 11 g/dL. Iron deficiency anemia (IDA) was defined according to ESMO 2018 Guidelines. The prevalence of ID, anemia and IDA was estimated using Agresti-Coull 95% confidence interval. Results: Patient’s median age was 63 years (55% women and 45% men). 89.4% of patients were currently under treatment for their cancer, mainly by chemotherapy (75.4%). The proportion of cancer patients with ferritin level < 100 μg/L was 20.5% [18.3-22.9] and 50.6% [47.8-53.4] of patients had a TSAT index < 20%. Overall, ID was found in 57.9% [55.1-60.6] of patients, with absolute ID accounting for 36% of all ID cases. 36.0% of patients were anemic and among them, IDA was reported in 21.8% [19.6-24.2] of patients. The type of cancer treatment did not influence the ID status as ID was also present in 57.4% of untreated cancer patient. Conclusions: This study highlights the high prevalence of ID in cancer patients, whether or not concomitantly associated with anemia or treatment. These results emphasize the need to better management of ID in cancer thereby optimizing overall patient care. Clinical trial information: NCT03924271.
Comparison of the tumor cells proliferative activity in donor tissue and samples of xenograft generations.
Authors: Lukbanova Ekaterina A. , Kolesnikov Evgeniy N. , Kozhushko Mikhail A , Kit Sergei , Myagkov Roman E. , Anisimov Aleksandr E. , Goncharova Anna S. , Protasova Tatiana P. , Karnaukhov Nikolay S. , Zaikina Ekaterina V. , Volkova A. V. , Mindar M. V. , Tkachev Sergey Yu. , Khodakova D. V. , Maksimov Aleksey Yu. , Vladimirova Liubov Yu
Published: May 2020

e15619

Background: Esophageal cancer is a common tumor of the mucous layer of the esophagus and the sixth most common cause of cancer deaths worldwide. Studying the mechanisms of its occurrence and development and the search for new approaches to its prevention and treatment are relevant. Such studies require a reliable experimental basis with such an important component as the use of animal models of cancer. In vivo models are an important tool both for testing new drugs and for studying the mechanisms of disease development. The purpose of the study was to create a patient-derived xenograft (PDX) model of esophageal cancer and to evaluate the proliferative activity of cells in the donor tumor and in xenograft samples in a series of successive generations. Methods: A PDX model of esophageal cancer was created by transplanting a tumor fragment from a patient with esophageal squamous cell carcinoma to the distal esophagus of BALB/c Nude athymic mice. The tumor histotype was confirmed histologically (hematoxylin and eosin staining). The proliferative activity of cells was evaluated by immunohistochemistry (IHC) with anti-Ki-67 antibodies. Results: Histological identity of xenograft samples and the donor tumor was confirmed. IHC showed an increase in the proliferative activity index in samples of transplanted tumors compared to the donor material: 12.4% in the primary tumor and 58%, 65.2%, 54.2% and 60.7% in generations 1, 3, 4 and 5, respectively. Conclusions: Histological characteristics of the donor tumor are preserved in all PDX models. The study demonstrated an increase of the proliferative activity in the first PDX generation compared to the primary tumor which persisted in the subsequent models.
Bstronximab, a novel therapeutic antibody to treat metastatic testis cancer.
Authors: Schopperle William Michael
Published: May 2020

e15621

Background: The treatment of metastatic testis cancer is by far the gold standard for treating metastatic solid cancers: over 90% of men diagnosis with metastatic testis cancer are completely cured with current therapies. What seems to be lost and somewhat surprising within this truly remarkable therapeutic cancer success in treating a metastatic cancer is the fact that in United States alone there are still over 400 young men who died from this disease because they do not respond to current therapies. Thus, despite the tremendous clinical success with treating this disease, there is still a strong unmet need to develop new treatments for men with metastatic testis cancer who fall outside the celebration of this disease as being completely cured. Methods: The human germ cell tumor embryonal carcinoma line, TERA-1. was used to generate purified testis cancer-specific cell membrane antigens to immunize mice and generated antibodies, by standard conventional mouse B cell hybridoma protocols. Isolated monoclonal antibodies where tested by ELISA and SDS-PAGE Western blotting to identify testis cancer specific antibodies. Testis cancer positive antibodies were further screen in an antibody-drug-conjugate screen to determine if the monoclonal antibody could function as a antibody-drug-conjugate therapeutic drug for metastatic testis cancer. Results: Dozens of monoclonal antibodies were generated with our antibody screen. Based on the screen results, a lead monoclonal antibody, named Bstrongomab, was selected for further development as a testis cancer therapeutic antibody. Bstrongomab is an IgG1 monoclonal antibody specific to the human embryonic cell marker, and testis cancer marker, TRA-1-60, a carbohydrate molecule which has been identified over 4 decades ago as a true human testis cancer marker. A human/mouse chimeric version of Bstrongomab was created - Bstrongximab - as a human therapeutic antibody. An antibody-drug-conjugate using Bstrongximab and the toxic payload MMAE (Monomethyl auristatin E) was developed and tested in pre-clinical human cancer testis in-vitro and in-vivo models and showed to be a powerful testis cancer therapeutic drug targeting TRA-1-60. Conclusions: Bstrongximab-MMAE is a novel therapeutic antibody-drug-conjugate which targets the testis cancer antigen TRA-1-60. TRA-1-60 is not expressed in normal tissues. Bstrongximab-MMAE is a new cancer drug ready to move into clinical trials for men with metastatic cancer that do not respond with current therapy protocols.
Multi-analyte interrogation based treatment of advanced refractory cancers.
Authors: Akolkar Dadasaheb B , Crook Timothy , Patil Darshana , Ranade Anantbhushan , Bhatt Amit , Limaye Sewanti , Datta Vineet , Schuster Stefan , Sims Cynthe , Srivastava Navin , Fulmali Pradip , Fulmali Pooja , Devhare Pradip , Patil Sanket , Apurwa Sachin , Srinivasan Ajay , Datar Rajan
Published: May 2020

e15624

Background: Treatment of advanced refractory cancers face challenges in non-availability of systemic therapy regimens with evidenced benefit. Post failure of two to three lines of systemic treatments, patients with such cancers are usually considered for palliation or clinical trials. Prior attempts at label-agnostic treatment regimens (precision medicine) in such populations were largely based on a single-indication-single-drug paradigm which had limited application. We hypothesized that advanced refractory malignancies have latent vulnerabilities which can be identified by an integrational multi-analyte interrogation of the tumor, and can be targeted using patient-specific combination regimens to yield clinical benefit. Methods: Fresh tumor tissue and blood samples were obtained from 158 patients with solid organ cancers where the disease had progressed following failure of at least two lines of standard of care systemic treatment options. These samples were used for Encyclopedic Tumor Analysis (ETA) which interrogated gene mutations, gene overexpression, pathway dysregulation, immunohistochemistry as well as in vitro chemosensitivity profiling of viable tumor cells. Integration of datasets from the multi-analyte ETA was used to generate patient-specific therapy recommendations. Patients who received ETA-guided treatments were followed up and response to treatment was retrospectively evaluated from radiological scans. Results: All patients received ETA-guided individualized treatments which were combinations of cytotoxic, targeted and endocrine agents. No two patients received the same treatment regimen. Complete or Partial Response (CR or PR) was observed in 76 patients yielding an Objective Response Rate (ORR) of 48.1%. 67 patients showed Stable Disease (SD), thus yielding a Disease Control Rate (DCR) of 90.5%. Median Progression Free Survival (PFS) was 117 days (Range 27 – 379 days). There were no Grade IV therapy related Adverse Events or therapy related deaths. Conclusions: Viable efficacious combination treatment options can be made available for patients with advanced refractory malignancies via ETA, despite perceived non-availability or non-viability of standard of care treatment options.
Biosimilar trastuzumab active post-marketing surveillance real-world data update: Results from a patient support program for patients under treatment with the first biosimilar trastuzumab approved in Brazil.
Authors: Silva Allisson Monteiro da , Paulo Jaqueline Amália de , Watanabe Thais Tiemi , Dib Gislaine Villarta Capeleti
Published: May 2020

e15626

Background: In 2017, biosimilar trastuzumab (Zedora) became the first biosimilar trastuzumab approved in Brazil. In 2019, biosimilar trastuzumab early data from an active postmarketing surveillance program were presented in ASCO. Now, with an extended follow up and more patients included, we present updated data from the surveillance program. Methods: This is a prospective observational study based on a patient support program. HER2 + breast cancer (BC) who received prescription for biosimilar trastuzumab were invited to participate. After agreement of informed consent, they were followed by periodical phone calls after each infusion and up to 3 months after the end of treatment. Treatment related data and adverse events (AEs) were collected. Results: A total of 76 female patients with HER2 + BC were enrolled in the program between May 2018 and December 2019. 74 patients with median age 51 (range 30 to 80) received at least one biosimilar trastuzumab infusion. Out of these, 52 (70.3%) patients reported 385 AEs all-causality (Table 1). Regarding severity and expectedness, 344 (89.4%) were non-serious AE (243 expected / 101 unexpected) and 41 (10.6%) were serious AE (29 expected / 12 unexpected). Considering all serious unexpected AEs (12), 8 were assessed as unlikely and 4 as likely. For the non-serious unexpected AEs (101) causality assessment was unlikely in 32 AEs, likely in 68 AEs and conditional in 1 AE. The three most frequently reported AEs according to SOC (System Organ Classification) were gastrointestinal disorders 59 (15.3%), general disorders and administration site conditions 57 (14.8%). The five most reported symptoms (PT - Preferential Term) were nausea 15 (3.9%), fatigue 14 (3.6%), infusion reactions 14 (3.6%), diarrhea 9 (2.3%) and insomnia 9 (2.3%). Further monitoring is continued. Conclusions: Nature of AEs for biosimilar trastuzumab are consistent with the label in female patients with HER2 + BC. The risk benefit remains consistent with the label and no new safety signals were detected. [Table: see text]
Injection mixed drug intratumoral to produce ROS and creating a coagulum as cancer therapeutics drug depot.
Authors: Zhang Jian , Fu Qiang , Li Bian , Han Yan , Chen Dong , Zheng Guoqin , Zhao Ming , Yu Baofa
Published: May 2020

e15635

Background: To explore the nature of hydrogen peroxide at higher dosage for denature-aggregation of tumor with drugs through aggregation in denatured tumor as a coagulum for drug depot and prolong function of drug. Methods: (1). Preparation for two 5 ml of BLM-I131, one diluted with 0.12 ml NS and 0.2 ml (0.2mCi = 49.67µg BLM) for tumor injection in control group; one diluted with 0.12 ml of H2O2 (0.833 mg/ml) and 0.2 ml (0.2mCi = 49.67µg BLM) for tumor injection in experimental group. (2).Tumor imaging with BLM-I131 analyzed at different time points. (3). Radioactivity in tumor of mice analyzed under SPECT scanning imaging instrument at the 0, 5 h, 1 h, 2 h, 4 h, 8 h, 24 h, 48 h, 96 h, 120 h, 144 h and 168 h, activity of isotope I131 is representing the BLM retaining time in tumor. (4). Also, tumor sectioned and observed cellular and extracellular matrix changes of histological structure. Results: It was observed that BLM-I131 with ROS in tumors sustained for 168 hours while BLM-I131 with NS in tumors sustained for 8 hours only. Radioactivity of BLM-I131 in tumors with ROS reach at peak 1.5 hours and second peak at 20 hours by average to extend to 168 hours, while radioactivity of BLM-I131 in tumors with NS reached at peak in 0.5 hour, decreased to 30% in 1.5 hour quickly to background in 8 hours; We observed the extracellular matrix changes in experimental tumor while no changes in control tumor. Conclusions: A water soluble oxidant mixed with free drug can play a biological scissors role to chop tumor matrix, then it resulted in a denature tumor matrix into a coagulation for a drug depot, it showed drug of BLM-I131 sustained in tumor for a long time; while oxidant plays an important role to punch holes on cell membrane and resulted a high permeability for high concentration drug in each cancer cells.
Effect of inhibition of receptor tyrosine kinase AXL by a selective small molecular inhibitor R428 (BGB321) on DNA damage repair response in ovarian cancer cells.
Authors: Huang Ruby Yun-Ju , Yeo Xun Hui , Tam Wai Leong
Published: May 2020

e15640

Background: AXL is a receptor tyrosine kinase that is often overexpressed in many cancers. It contributes to tumor progression, metastasis and drug resistance through activating downstream signaling cascades, making it an emerging therapeutic target. The first-in-class AXL inhibitor R428 (BGB321) was approved by the FDA for the treatment of relapsed or refractory acute myeloid leukemia. R428 (BGB321) was also reported to show selective sensitivity towards ovarian cancers (OC) with a Mesenchymal (Mes) molecular subtype. Recently, a novel role of AXL in the regulation of DNA damage responses has been described. In this study, we explored further the role of AXL in mediating DNA damage responses by using OC as a disease model. Methods: OC cell lines were treated with R428. Accumulation of γH2AX positive foci was assessed for DNA damage response. Western blotting for γH2AX, ATM and ATR levels were performed. Dose response curves of ATR inhibitors were generated by treating OC cells with the fixed dose of R428 (IC20 concentration of each cell line). Results: AXL inhibition by using R428 resulted in the increase of DNA damage foci in Mes OC cells SKOV3 and HeyA8. This occurred concurrently with the up-regulation of classic DNA damage response signaling molecules such as γH2AX, ATM and ATR. The IC50 of the ATR inhibitor significantly decreased for 2-3 folds in all OC cell lines tested. AXL inhibitor R428 sensitized both BRCA-mutated and non-BRCA-mutated OC cells to a potent and highly selective ATR inhibitor. Conclusions: Our results showed that AXL inhibition rendered cells more sensitive to the inhibition of ATR, a crucial mediator for replication stress, paving ways to the rationale for potential combinatory use of AXL and DNA damage repair inhibitors.
Next generation proteomics in precision oncology: 1000s of proteome and phosphoproteome profiles of tumors and matching healthy tissues as meaningful layer in multi-omics database.
Authors: Escher Claudia , Vowinckel Jakob , Novy Karel , Corwin Thomas , Treiber Tobias , Bruderer Roland , Reiter Lukas , von Leitner Eike-Christin , Rinner Oliver
Published: May 2020

e15672

Background: The rise of precision oncology therapeutics requires deep understanding of all molecular mechanisms involved in cancer biology. IndivuType offers the world’s first multi-omics database for individualized cancer therapy, analyzing the highest quality cancer biospecimens to generate the most comprehensive dataset, including genomics (WGS), transcriptomics, proteomics, and clinical outcome information. Indivumed is committed to the quality of the IndivuType ecosystem starting with stringent SOP-driven sample collection combined with thorough validation of clinical information and data integrity. The availability of multi-omics data from the same tumor can provide a comprehensive molecular picture of cancer for a given patient. Protein expression and activation are directly related to cellular function and hence provide actionable information about druggable targets. Until recently, the proteomics technology could not match the scale of next-gen sequencing and consequently precision medicine has almost exclusively been based on gene level data. Here we present the first large-scale data set for protein expression and phosphorylation. Enabled by the data independent acquisition (DIA) workflow, a mass spectrometric method provided by Biognosys that obtains peptide fragmentation data in a highly parallelized way with high sensitivity, more than 7,000 proteins in the whole proteome (WP) and 20,000 phospho-peptides in the phospho-proteome (PP) workflow were profiled. Methods: Sample processing from 5 mg of tissue per sample was performed using liquid handling robot. Phospho-peptide enrichment was carried out with a Kingfisher Flex device and MagReSyn Ti-IMAC magnetic beads. DIA LC-MS/MS was performed on multiple platforms consisting of a Thermo Scientific Q Exactive HF-X mass spectrometer coupled to a Waters M-Class LC. Chromatography was operating at 5 µL/min, and separation was achieved using 45 min (WP) and 60 min (PP) gradients. Results: Several thousands of high-quality patient samples of various cancer types have been analyzed to date. The resulting proteome and phospho-proteome data has been integrated into the IndivuType database, thereby providing a solid foundation to advance our understanding of cancer. Conclusions: With the ongoing addition of more samples and associated deep and rich data, the platform could unravel key molecular events and is expected to transform knowledge into actionable treatments and personalized therapies.
Immunohistochemical biomarker expression in gastro-entero-pancreatic neuroendocrine tumors at ENETS Centre of Excellence, University Hospital Zurich, Switzerland.
Authors: Duschek Silvia , Friemel Juliane , Curioni Fontecedro Alessandra , Siebenhuener Alexander Rheinhard
Published: May 2020

e15673

Background: Gastro-entero-pancreatic neuroendocrine tumors (GEP-NET) are a heterogeneous tumor entity with respect to biological behaviour and prognosis. Definition of new predictive and prognostic biomarkers as well as new therapy targets are of upmost clinical interest. Methods: At the ENETS CoE Zurich tissue microarray (TMA) blocks were constructed with 147 tissue samples of tumors diagnosed from 2000 to 2017, including primary tumors and metastasis. Tissue microarray sections were immunostained for SOX-9, SOX-10, SSTR-2 +, PDL-1, thyroid transcription factor 1 (TTF1), estrogen receptor-α (ER-α) and -β (ER-β), progesterone receptor (PR), androgen receptor (AR), BRAF and HER2. Results: In total 270 patients were screened between 2000 - 2017 and 92 patient’s material were sufficient for TMA analysis. Subgroup analysis showed 32 pancreatic, 37 ileum, 10 duodenum, 7 appendix, 3 colorectal, 3 gastric and 1 NET of gallbladder. 50% were male, the median age 63.5 years (range 19-88y). AR, ER-β, TTF1, PDL-1 and SOX10 was only present in a small number of NETs. HER2 and BRAF were negative in all samples. We found ER-α expression in 27.2%, 30% were PR positive and 67.4% showed SOX9 expression. Conclusions: We identified a frequent expression of new markers as SOX9, progesterone receptor and estrogen receptor-α in a broad amount of samples and a potential correlation with tumor grade. This finding might implicate a prognostic or predictive value of these markers, as well as it reveals new potential therapy targets for GEP-NETs. Therefore, further analyses are planned.
Expression profile and potential regulatory pathways of SFTPA1 in human tumors.
Authors: Yuan Lu , Yang Mi , Wu Xixi , Huang Wenqi , Wu Yuting , Hwang Jihong , Guan Jian , Liu Laiyu
Published: May 2020

e15675

Background: Pulmonary surfactant protein A1(SFTPA1) plays a critical role in maintaining lung tissue homeostasis and its disruption could cause lung diseases and even lung cancer. Despite the finding that SFTPA1 is a potential biomarker for lung cancer, the expression profile and molecular mechanism of SFTPA1 in pan-cancer remain unclear. Methods: The expression of SFTPA1 in BALB/c mice, human normal tissues and human tumor tissues were explored by immunohistochemistry assay. The prognostic values were evaluated by Kaplan-Meier Plotter Database Analysis. Principal components analysis and GO and KEGG pathway enrichment analyses were exerted to identify the functional heterogeneity of SFTPA1 in various tumors. Multiple databases were used to analyze immune infiltrates and regulation networks of SFTPA1. Results: This study described the expression profile of SFTPA1 in various organs and tissues of BALB/c mice and various types of human tumor tissues. Interestingly, we found the expression level of SFTPA1 mRNA was associated with different prognosis in different cancers, even in Lung adenocarcinoma(LUAD) and Lung squamous cell carcinoma(LUSC). The further analysis confirmed a functional heterogeneity of SFTPA1 across diverse cancer types. Significantly, SFTPA1 might regulate tumor microenvironment by affecting the polarization of M2 macrophages in LUAD and LUSC. Furthermore, we found the has-miR-3200-3p-HNF1B-SFTPA1 network was a potential upstream regulatory mechanism of SFTPA1 in LUSC. Conclusions: Our study elaborated expression profile and potential regulatory pathways of SFTPA1 in human solid cancers, and might provide a potential immune infiltration biomarker for lung cancer.
Neutrophil extracellular traps blockade in combination with PD-1 inhibition in treatment of colorectal cancer metastasis.
Authors: Onuma Amblessed , He Jiayi , Xia Yujia , Zhang Hongji , Genkin Dmitry , Tetz George , Huang Hai , Tsung Allan
Published: May 2020

e16002

Background: Immune checkpoint inhibitors (ICIs) are currently approved for microsatellite instability-high metastatic colorectal cancer (mCRC) however reported objective response rate to monotherapy range between 28-52%. Neutrophil Extracellular Traps (NETs), an extracellular network of DNA and proteins expelled by neutrophils into the tumor microenvironment, promote CRC metastasis by inducing potent immunosuppressive effects. We hypothesize that targeting this network may improve response rates to immune checkpoint therapy. The purpose of our study is to investigate the effect of combination DNase I treatment, a NET depleting agent, and PD-1 blockade in mCRC progression. Methods: Subcutaneous MC38 (MSI-H adenocarcinoma of the colon) tumors were established in 6-week-old mice. Treatment was initiated six days after tumor inoculation with daily intraperitoneal (i.p) injections of DNase I followed by i.p anti-PD-1 treatment administered every 3 days. Animals were divided into treatment groups: control, DNase alone, anti-PD-1 alone and combination of DNase I with anti-PD-1 (n = 6 mice/ group). Results: Tumors were successfully established after 6 days of inoculation. These tumors expressed high neutrophil infiltration and NETs. To determine the effects of combination therapy on tumor progression, we measured tumor volume and time to progression (TTP). We found that mice treated with combination DNase I and anti-PD-1 had significantly lower mean tumor volume compared to anti-PD-1 alone (1372.68mm3 vs 2193.20 mm3, p-value = 0.043). TTP was higher in the combination group compared to anti-PD-1 treatment alone (median time to progression 22 vs 17 days, p-value = 0.004). Next we sought to determine the mechanism behind this effect by investigating if NETs blockade modulated the tumor microenvironment. We found that treatment with DNase I decreased exhaustion of CD8+ T-cells in the tumor compared to no treatment. Conclusions: Targeting NETs in combination with immune checkpoint inhibition may be an option to improve response rate to immune checkpoint inhibitors through decreasing exhausted CD8+ T-cells.