Dessie, Ethiopia

A Prospective Study of Liquid Biopsy for Pancreatic Cancer Early Detection

Peripheral blood samples from participants with new diagnosis of pancreatic cancers will be collected to characterize the cancer-specific circulating signals by sequencing cell free DNA. A noninvasive test integrating machine learning algorithm will be trained and validated through a two-stage approach in recruited well-classified individuals, along with non-cancers without clinical diagnosis of cancer after routine medical screening. The performance of liquid biospy assays discovering cancer from non-cancer will be evaluated in participants with benign disease as well as average risk individuals.

AssesSment of Early deteCtion basEd oN liquiD Biopsy in Intracranial Tumors

Assessment of Early-detection Based on Liquid Biopsy in Lymphoid Malignancies

Fluid Biopsy for the Diagnosis of Lung Cancer

PRIMARY OBJECTIVE: I. To estimate the sensitivity and specificity of the fluid biopsy in establishing a diagnosis of bronchogenic carcinoma. SECONDARY OBJECTIVES: I. To determine the accuracy of determination of the histologic subtype of bronchogenic lung cancer. II. To determine the relative contribution of cell based high definition circulating tumor cell assay (HD-CTC) and imaging mass cytometry (IMC) and plasma based circulating tumor deoxyribonucleic acid (ctDNA) assays in determination of diagnosis and histologic subtype. OUTLINE: Patients undergo collection of blood samples on day 1 for analysis via high definition (HD)-single cell analysis (SCA) fluid biopsy. Medical charts of patients are reviewed at 3 months post-biopsy or computed tomography (CT) screening. After completion of study, patients are followed for up to 1 year.

Liquid Biopsy Assay of EBUS-TBNA Supernatant Fluid for Diagnosing Lung Cancer

Background Lung cancer is the leading cause of cancer deaths. In particular, non-small cell lung cancers (NSCLC) accounts for over 80% of cases. Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is the first-choice procedure for staging of NSCLC, as it allows a low-risk, minimally invasive histopathological tumor diagnosis, and provides adequate material for immunohistochemistry staining and molecular testing. During the procedure, thoracic lymph nodes are sampled under endobronchial sonographic guidance. However, 10-15% of EBUS-TBNA does not yield adequate cytological material for precise diagnosis, staging, or further testing. This may result from sampling errors, low quality or quantity of collected material, or abundance of necrosis. In those cases, further invasive surgical procedures such as mediastinoscopy or thoracoscopy may be considered to complete the staging determination. Next generation sequencing (NGS) of tumor cells is recommended to identify molecular alterations to select targeted therapies and improve outcomes. When tissue samples are limited or non-diagnostic, NGS can also be performed on peripheral blood or other body fluids, such as malignant pleural effusions and EBUS-TBNA supernatant, by liquid biopsy (LB) technics. Molecular profiling of peripheral blood by LB is highly concordant with NGS of tissue biopsies, the same mutations are detected by both methods, and is thus gaining an increasing popularity in the management of patients with NSCLC. LB can provide a molecular characterization when tissue quality or quantity is not sufficient, identify possible therapeutic targets when such are not detected by tissue NGS, and guide further treatment in cases of disease progression. LB NGS of EBUS-TBNA supernatant was assessed in several small-scale studies, which found good concordance with NGS of tissue biopsies and peripheral blood, improved failure rates, and possibly reduced costs and turnaround times. Those studies included supernatant from positive procedures only, that is, procedures in which tumor cells were identified from EBUS-TBNA samples. Currently, LB NGS cannot ascertain (or rule-out) lymph nodes involvement in tumor development, and thus cannot contribute to the staging process. This study aim is to preliminary assess whether LB NGS from the supernatant could contribute to identifying lymph nodes involvement in subjects diagnosed with NSCLC, but determined as negative for lymph nodes involvement by EBUS-TBNA. The study hypothesis is that there is an excellent correlation between LB NGS from EBUS-TBNA supernatant and tumor involvement of the lymph nodes, even when such involvement is unrecognized by EBUS-TBNA cytology. Study Objectives General Aim To preliminary assess whether LB NGS of the supernatant may be indicative of lymph node involvement in patients with non-small cell lung cancer (NSCLC) in whom no tumor cells have been identified in the aspirate (false negative). Specific Aims 1. To assess the concordance between the positivity of supernatant LB NGS and surgical lymph nodes staging in subjects with a negative EBUS-TBNA evaluation (no tumor cell detected) who will require surgical resection of the thoracic lymph nodes. 2. To assess the concordance between LB NGS of the supernatant and NGS of surgical tumor specimens. 3. To assess the concordance between LB NGS of the supernatant and LB NGS from peripheral blood. Methods This is a prospective feasibility study. The primary outcome will be the concordance between the results of EBUS-TBNA supernatants NGS (positive vs. negative for any mutation commonly associated with NSCLC) and the final pathological staging following surgery (positive vs. negative thoracic lymph nodes involvement). Secondary exploratory outcome will be the concordance between genetic alterations detected by comprehensive tissue NGS and those detected by supernatant LB, cell-free DNA NGS, and the concordance between peripheral blood and supernatant LB NGS. Inclusion Criteria: 1. Ten adult (age ≥18 years) patients with high suspicion for NSCLC with thoracic lymph nodes involvement (per clinical judgement and imaging studies), planned for an EBUS-TBNA procedure, will be recruited. 2. Subjects with negative EBUS-TBNA results (no evidence of lymph node involvement by tumor according to histology) who will require surgical resection of the thoracic lymph nodes will comprise the final study group. Exclusion Criteria: 1. Subjects unable or not willing to provide informed consent for study participation. 2. Subjects in whom NSCLC will ultimately be ruled out. 3. Subjects who will not require surgical resection of thoracic lymph nodes, or who will not undergo such procedure in our Medical Center for any reason. Sample Size: 10 participants will be recruited for this feasibility study. This should allow detection of a significant agreement (Cohen's Kappa) of >0.55, with 80% power and type I error (alpha) of 0.05. Measured Variables: Collected data will include: - Age - Gender - Smoking history - Past medical history, co-morbidities, and medications - Imaging results and radiological staging - EBUS-TBNA report, including lymph nodes size and structure, number, and location of sampled nodes - Surgery reports - Final surgical pathology and staging - NGS of tumor tissue from surgical specimens - LB NGS from EBUS-TBNA supernatant - Treatment decisions Study Procedures: Participants will undergo EBUS-TBNA procedures under moderate sedation, as commonly practiced. During the procedures, suspicious thoracic lymph nodes will be sampled using sonographic guidance. EBUS-TBNA needle and syringe rinse fluid will be collected in tubes containing 30ml of CytoLyt (Hologic, Marlborough, Massachusetts). Samples will be centrifuged at 1200 RPM for 10 minutes, the supernatant will be poured into another tube, and the cell pellet will be fixed in formalin and embedded in paraffin as a cell block, for standard cytopathological workup. The supernatant (which is usually discarded) will be stored at 4°C until surgical resection of the lymph nodes will be recommended by a Tumor Board. DNA and RNA will be extracted from formalin-fixed and paraffin-embedded (FFPE) tissue for NGS using Oncomine™ Comprehensive Assay Plus for therapeutic decisions as generally practiced. For the subjects in the final study group, cell-free DNA will be extracted from the supernatant samples and NGS will be performed using Oncomine™ Pan-Cancer Cell-Free Assay, and also from peripheral blood, according to manufacturer recommendations and as previously described. Both assays can detect a wide spectrum of genetic mutations which are typically associated with NSCLC. All extractions, sequencing, analysis, and calculation will be performed in the Pathology Laboratory of Barzilai University Medical Center. Statistical Analysis: Descriptive statistics will include mean and medians with standard deviations and range, absolute numbers, and percentages. Agreement between tests will be calculated using Cohen's Kappa statistic.

Liquid Biopsy to Enable Diagnostics and Monitoring for Immune-mediated Lymphoproliferative Disorders

In this observational prospective study, the investigators will collect clinical data from subjects' charts through a dedicated multicenter electronic case report form (eCRF). Whenever available, PET-CT will be transferred through a Web-based Imaging and Diagnosis Exchange Network (WIDEN) to perform a blinded independent review of staging and response. Biological samples included 20 ml of blood collected at each of the following planned clinical points in time: i) at ILD diagnosis, ii) after first cycle of therapy, iii) at interim response assessment, iv) at the end-of-treatment, v) at 3 months follow-up, vi) at 12 months follow-up, vii) at disease progression, if applicable. Additional samples could be collected if clinically relevant. Additionally, the investigators will request formalin-fixed and paraffin-embedded tissue (FFPET) or fresh-frozen (FF) tissue slices/blocks at ILD diagnosis from Pathology Departments of participating Centers for retrospective ILD subjects.

Liquid Biopsy and Pancreas Cancer: Detection of AXL(+) CTCs (CTC-AXL-PANC)

In solid cancers, after the formation and growth of the primary tumor, some more aggressive tumor cells actively detach from it and then travel through the circulating compartment to reach distant organs (bone marrow - liver - lung - brain…) and constitute new foci or micro-metastases. These circulating tumor cells (CTCs) that have become disseminated tumor cells (DTCs) flourish in their new environments and may remain dormant for many years after the complete resection of the primary tumor. Due to events not fully elucidated, DTCs can develop on site giving rise to macroscopic metastases but also join again the circulating compartment in the form of CTCs, swarm, colonize other organs and cause secondary metastases. Detecting CTCs in the blood is very relevant for assessing tumor progression but also promising in terms of cancer disease prognosis and therapeutic follow-up. This new approach, published for the first time in 2010 under the term liquid biopsy, is therefore defined as a non-invasive blood test, extremely sensitive, achievable in real time and that allows the analysis of CTCs. Currently, the choice of targeted therapies for a given patient is made after analyzing the primary tumor for expression and/or genomic status of specific molecular targets. Many studies show that metastatic cells have phenotypic and genotypic characteristics distinct from those of most of the primary tumor. This can be explained either because metastatic cells acquire new genomic skills over time, or because a subset of metastatic sub-clone pre-exists within the primary tumor but has escaped detection by standard tissue biopsy techniques. A direct analysis of CTCs could provide important additional information to prevent patients from inappropriate, costly treatments and harmful side effects. For several years, the AXL protein, a tyrosine kinase receptor, has emerged as a new strategic target in oncology. Over-expression of AXL has been frequently identified in patients with pancreatic adenocarcinoma. AXL is a member of the Tyro3-Axl-Mer family, like its ligand protein Gas-6 (growth arrest-specific). An activation of the Gas6/AXL signaling pathway results in the activation of several effector pathways such as RAS/RAF/MEK/ERK or PI3K/AKT and is associated with, among other things, tumor cell growth and survival, metastatic formation and dissemination, Epithelial-to-mesenchymal transition (EMT) or drug resistances. It has been shown clinically that the AXL protein is a factor of poor prognosis and resistance to reference treatments (radiotherapy, chemotherapy or targeted therapy). Thus, many therapeutic strategies have been proposed and developed to inhibit the AXL pathway, ranging from chemical molecules, blocking its kinase activity and therefore the underlying signaling pathways, to nucleotide aptamers, AXL fusion proteins, and monoclonal antibodies. Pancreatic adenocarcinoma, the 4th leading of cancer related deaths, remains among cancers of very poor prognosis and thus represents a major therapeutic challenge. The median overall survival is 11.1 months after optimal treatment (FOLFIRINOX). The clinical relevance and oncogenic potential of AXL in the progression of different types of tumors have been largely evidenced. Indeed, 50% to 75% of pancreatic adenocarcinoma samples have overexpression of AXL and the level of expression of AXL is correlated with clinical parameters indicating tumor aggressiveness and poor prognosis such as frequency of distant metastases or the survival. In this context, the LCCRH lab, which has specialized in the detection and analysis of CTCs for 20 years, has developed a CTC-AXL detection test using the CellSearch® system. The CellSearch® system is the only method approved by the Food and Drug Administration (FDA) for the detection of CTC in colorectal, breast and metastatic prostate cancers. In addition, the LCCRH holds a patent for another technology for the detection and characterization of live and functional CTCs, called EPIDROP. The implementation of the AXL research is already done for AXL labelling on CTCs in EPIDROP as well as the visualization of the AXL cleavage by live CTC. Thanks to this unique functional test of CTCs, it is easy to imagine offering an oncology 'oncogram' by testing in real time the effectiveness of drugs on CTCs and personalized medicine to patients. This real-time liquid biopsy proposal on functional CTCs is quite innovative in Oncology. To date, there are no studies on the study of functional CTCs related to AXL. Primary objective: - Evaluate the concordance of the CTC-AXL measurement (inclusive) by the innovative EPIDROP technique and the CellSearch technique® Secondary objectives: - Evaluate the accuracy of the CTC-AXL measurement (inclusive) between EPIDROP and CellSearch® - Assess the degree of agreement between CTC-AXL measurement (inclusive) by EPIDROP and CellSearch® - Evaluate the overall survival of patients with metastatic pancreatic cancer based on the number of circulating tumour cells carrying the AXL marker measured by EPIDROP or CellSearch® at inclusion - Evaluate the progression-free survival of patients with metastatic pancreatic cancer based on the number of circulating tumour cells carrying the AXL marker measured by EPIDROP or CellSearch® at inclusion - Culture CTCs from the blood sample (EDTA 10mL) - Creation of a single liquid biopsy bio-bank for pancreatic cancer involving only plasma storage - Study the expression of PD-L1 (at inclusion) and detect the CTC subgroup expressing PD-L1 using EPIDROP and CellSearch® techniques; - Study the circulating immune system in this cohort of patients with metastatic pancreatic cancer by performing immunological analysis on blood mononuclear cells; - Determine if there is a significant correlation between the detection of CTC and CTC-PD-L1 and the circulating immune system (T cells, NK cells, B cells, macrophages, immune checkpoints, platelets); - To assess the correlation between CTC, CTC-PD-L1 and immune cells and the clinical outcomes (progression-free survival, overall survival) of these patients with metastatic pancreatic cancer.

International Study of High-Risk Patients for Hepatocellular Carcinoma Using Liquid Biopsy

Vietnam and Saudi Arabia have some of the highest disease burdens of liver cancer globally. Early detection in asymptomatic patients who are at risk for liver is a strategy to improve survival outcomes in liver cancer management. GALAD score (gender, age, alpha-feto protein (AFP)-L%, AFP and DCP) is a serum biomarker-based panel that can improve HCC early detection in patients with liver fibrosis and cirrhosis. In case-control studies and studies with the design of prospective specimen collection, and retrospective blinded evaluation, GALAD has demonstrated promising clinical utility. However, in order to ascertain its potential role in the surveillance of liver cancer early detection, GALAD needs to be validated prospectively for clinical surveillance of liver cancer (i.e. phase IV biomarker validation study). Thus, the investigators propose to conduct a phase IV biomarker validation study to prospectively survey a cohort of patients at risk for HCC (i.e. patients with compensated cirrhosis and irrespective of cirrhosis etiologies), using semi-annual abdominal ultrasound and GALAD Score for 5 years. In doing so, the investigators aim to validate the potential role of GALAD Score for clinical surveillance and early detection of HCC in Vietnam and Saudi Arabia. Additionally, the investigators will collect and archive biospecimens to develop a bio-repository for liver disease. The biorepository will encourage the sharing of biospecimens and collaboration among physicians or physician-researchers between the US, Vietnam, and Saudi Arabia.

Liquid Biopsy of Head and Neck Cancer Patients in Blood and Saliva

Clinical examination including imaging and - if necessary - tissue biopsy sampling - is the current clinical standard in therapy monitoring of metastatic head and neck tumors. This includes both the initial diagnosis, the assessment of the therapeutic response during ongoing chemotherapy / radiochemotherapy and follow-up care with the aim of detecting recurrences at an early stage. The detection of circulating nucleic acids as well as proteins in the peripheral blood and saliva could represent a minimally invasive and exact method for the assessment of the tumor burden, for the early detection of recurrences and for the individual assessment of the therapy response in patients with head and neck cancer. The present study aims to evaluate the value of tumor-specific nucleic acids and proteins in peripheral blood and saliva as possible biomarkers for minimally invasive therapy monitoring of head and neck tumors. For this purpose, Next Generation Sequencing (NGS), ELISA and quantitative polymerase chain reaction (PCR) methods are used as diagnostic methods. NGS initially enables the creation of a genetic profile of the primary tumor with targeted massive parallel sequencing of frequently mutated genes in head and neck tumors. The amount of nucleic acids in the peripheral blood and saliva is then quantified by means of digital PCR with the aid of specifically designed digital PCR assays. In addition, tumor-associated nucleic acids and proteins in the primary tumor, blood and saliva are examined. The aim is to examine if the amount of tumor specific circulating nucleic acids and the concentration of protein biomarkers found in the blood and saliva are associated with the response to treatment, early detection of recurrence, and the overall prognosis.

Monitoring Minimal Residual Disease in Gastric Cancer by Liquid Biopsy Study Description

Gastric cancer is the fourth most common cancer in Vietnam with high mortality rate. Patients at early stages undergo radical gastrectomy with curative intent, but the remaining tumor cells, termed as minimal residual disease (MRD), can later cause relapse. Conventional methods to monitor MRD such as imaging and blood tests for biomarkers such as CEA are not sensitive and specific enough. ctDNA has recently emerged as a promising noninvasive marker with high accuracy to monitor MRD and detect relapse in many cancers such as breast and colorectal cancers. However, its application in gastric cancer has not been extensively evaluated. Therefore, this study aims to use advanced NGS technologies to detect ctDNA in liquid biopsy as a biomarker to monitor MRD after radical gastrectomy.