Day 1 :
- Hematology- Oncology
Location: Webinar
Session Introduction
Amna Qamar Uz Zaman
Dr. Ziauddin University and Hospital, Pakistan
Title: Positron emission tomography – computed tomography as a monitoring response tool in prefibrotic myelofibrosis: A case report
Biography:
Dr Amna has done her MBBS in 2010 from Dow University Of Health Sciences and FCPS (Hematology) in 2018 from Dr. Ziauddin University Hospital.
Since 2018 I have been serving in the department of Hematology/Oncology diagnosing and treating a number of benign and malignant disorder including nutritional deficiencies, acute leukemia, chronic leukemia, lymphoma, hemoglobinopathy, inherited bone marrow failure syndromes, coagulopathies, transfusion related problems, thrombophilia etc.
Abstract:
Primary myelofibrosis is a haematopoietic stem cell neoplasm resulting in ineffective haematopoiesis and bone marrow fibrosis. We present a case of a 67-year-old male patient who came to the oncology/haematology department of Dr. Ziauddin Hospital, Karachi, in February 2020 with complaints of weight loss, gastroesophageal reflux and loss of appetite. Examination revealed splenomegaly and initial workup demonstrated bicytopenia on complete blood picture. Bone marrow biopsy was consistent with pre-fibrotic myelofibrosis (Janus kinase 2 (JAK-2) positive). He was categorized as intermediate-2 risk according to Dynamic International Prognostic Scoring System (DIPPS) with score of 3 and was advised to start JAK-1/JAK-2 inhibitors. Prior to therapy, he underwent positron emission tomography-computed tomography (PET/CT) scan which showed increased fluorodeoxyglucose (FDG) uptake in the spleen and bone marrow. Monitoring by the scan after initiating treatment demonstrated decreased FDG uptake in bone marrow and spleen, demonstrating that PET/CT is a non-invasive way to assess and monitor treatment response in pre-fibrotic myelofibrosis.
Biography:
Mengying(Mona) Liu has her expertise in molecular biology, microbiology, cell biology, gene & cell therapy, drug development, and biomarker discovery. She has experience in process and assay development. Current research focuses on assay development to provide accurate and high quality IVT products for cancer diagnostic and monitoring.
Abstract:
Statement of the Problem: BCR-ABL p190 (e1a2) is expressed in 20-30% of Acute Lymphoblastic Leukemia (ALL) and 1-2% of Chronic Myeloid Leukemia (CML)1,2. The co-expression of p190 and p210 is rare in both CML and ALL1,2. Xpert® BCR-ABL Ultra p190, an automated cartridge-based test for measuring BCR-ABL p190 transcript level, is standardized to quantify the amount of BCR-ABL p190 relative to ABL1 control gene based on delta Ct in peripheral blood of patients3. Since BCR-ABL p190 level is crucial for risk stratification and treatment decisions in ALL4 as well as diagnosis and continuous therapeutic monitoring in CML5, it can be useful to obtain the copy number (CN) of BCR-ABL p190. The purpose of this study is to establish a BCR-ABL p190 CN estimator with known CN of e1a2-ABL IVT-RNA as well as to compare %BCR-ABL p190/ABL1 reporting between delta Ct-based and CN-based methods. Methodology & Theoretical Orientation: Nine levels of e1a2-ABL IVT-RNA as well as two lots of Xpert® BCR-ABL Ultra p190 tests were used to generate standard curves for CN and %CN reporting. BCR-ABL p190 contrived samples and ALL clinical samples containing the BCR-ABL p190 transcript were examined to evaluate the CN and %CN between two lots of the Xpert® BCR-ABL Ultra p190 tests and to compare the delta Ct-based and CN-based methods for reporting %BCR-ABL p190/ABL1. Findings: Linearity was demonstrated in Ct vs CN input for BCR-ABL p190 (R2>0.99) and ABL1 (R2>0.98). Less than 1.35-fold difference was exhibited for CN and %CN across two different lots. Less than 2-fold difference was observed in %BCR-ABL p190/ABL1 reporting between delta Ct-based and CN-based approaches. Conclusion & Significance: A BCR-ABL p190 copy number estimator for Xpert® BCR-ABL Ultra p190 test was established, which will provide helpful information for diagnosis and prognosis of diseases relating to BCR-ABL
Biography:
(Amber) Zhao has her expertise in molecular biology, microbiology, and cell biology. She has experience in process and assay development. Current research focuses on assay development to provide accurate and high quality IVD products for cancer diagnostic and monitoring.
Abstract:
Statement of the Problem: The nucleophosmin (NPM1) is the most mutated gene (~30%) in Acute Myeloid Leukemia (AML)1. Three NPM1 mutations (type A, B, and D) represent ~84% in NPM1-mutated AML cases while other uncommon subtypes occupy ~16%2. Xpert® NPM1 mutation, an automated cartridge-based test for measuring NPM1 mutation transcript levels (type A, B and D), is standardized to quantify the amount of mutated NPM1 relative to ABL1 control gene based on delta Ct in peripheral blood3. Since mutated NPM1 level is crucial for risk assessment, medication selection, and ongoing therapeutic monitoring in AML4,5, it is important to obtain the NPM1 mutation copy number (CN). The aim of this work is to develop NPM1 mutation CN estimator and to compare %NPM1 mutation/ABL1 reporting between delta Ct-based and CN-based methods. Methodology & Theoretical Orientation: Five levels of NPM1 mutations (A, B, D) and ABL1 IVT-RNA panels as well as two lots of Xpert® NPM1 mutation tests were used to generate standard curves for CN and %CN reporting. The cell lysates from cell lines carrying either NPM1 mutation A, B, or D and AML clinical samples containing NPM1 mutations were examined to evaluate the CN and %CN between two lots of the Xpert® NPM1 mutation tests and to compare the delta Ct-based and CN-based methods for reporting %NPM1 mutation/ABL. Findings: Linearity was demonstrated in Ct vs CN input for NPM1 mutation and ABL1 with R2 above 0.96 for Lot1 and Lo2. Less than 3-fold difference was exhibited for CN and %CN across two lots of Xpert® NPM1 mutation test. Less than 3-fold difference was observed in %NPM1 mutation/ABL1 reporting between delta Ct-based and CN-based approaches. Conclusion & Significance: An NPM1 mutation copy number estimator for Xpert® NPM1 mutation test was established, which will provide diagnostic and prognostic values for NPM1-mutated AML patients.
Biography:
Mengying(Mona) Liu has her expertise in molecular biology, microbiology, cell biology, gene & cell therapy, drug development, and biomarker discovery. She has experience in process and assay development. Current research focuses on assay development to provide accurate and high quality IVD products for cancer diagnostic and monitoring.
Abstract:
Statement of the Problem: BCR-ABL p190 (e1a2) is expressed in 20-30% of Acute Lymphoblastic Leukemia (ALL) and 1-2% of Chronic Myeloid Leukemia (CML)1,2. The co-expression of p190 and p210 is rare in both CML and ALL1,2. Xpert® BCR-ABL Ultra p190, an automated cartridge-based test for measuring BCR-ABL p190 transcript level, is standardized to quantify the amount of BCR-ABL p190 relative to ABL1 control gene based on delta Ct in peripheral blood of patients3. Since BCR-ABL p190 level is crucial for risk stratification and treatment decisions in ALL4 as well as diagnosis and continuous therapeutic monitoring in CML5, it can be useful to obtain the copy number (CN) of BCR-ABL p190. The purpose of this study is to establish a BCR-ABL p190 CN estimator with known CN of e1a2-ABL IVT-RNA as well as to compare %BCR-ABL p190/ABL1 reporting between delta Ct-based and CN-based methods. Methodology & Theoretical Orientation: Nine levels of e1a2-ABL IVT-RNA as well as two lots of Xpert® BCR-ABL Ultra p190 tests were used to generate standard curves for CN and %CN reporting. BCR-ABL p190 contrived samples and ALL clinical samples containing the BCR-ABL p190 transcript were examined to evaluate the CN and %CN between two lots of the Xpert® BCR-ABL Ultra p190 tests and to compare the delta Ct-based and CN-based methods for reporting %BCR-ABL p190/ABL1. Findings: Linearity was demonstrated in Ct vs CN input for BCR-ABL p190 (R2>0.99) and ABL1 (R2>0.98). Less than 1.35-fold difference was exhibited for CN and %CN across two different lots. Less than 2-fold difference was observed in %BCR-ABL p190/ABL1 reporting between delta Ct-based and CN-based approaches. Conclusion & Significance: A BCR-ABL p190 copy number estimator for Xpert® BCR-ABL Ultra p190 test was established, which will provide helpful information for diagnosis and prognosis of diseases relating to BCR-ABL p190.
Rachana Garg
Deapartment of Surgery, City of Hope Medical Center, California, USA
Title: CBX2: a potential marker of HER2+ breast-to-brain metastasis
Biography:
I, Rachana Garg, have been DoD postdoctoral fellow and Research Associate Scientist at the University of Pennsylvania, USA. Currently, I work as a Staff Scientist with City of Hope National Medical Centre, Duarte, California, USA. My experience spans the field of cancer biology, neuroscience, clinical research, drug discovery, chemoprevention cellular and molecular biology, biochemistry, immunology, epigenetics. During my academic career, I have received multiple awards at international conferences, and my research has been published in top journals: Cancer Research, Cell Reports, Oncogene, Carcinogenesis, Oncotarget, Journal of Biological Chemistry, Frontiers in Oncology, BBA Reviews in Cancer (1179 citations, H-index 14, & I-10 index 15). In addition to my passion and involvement in teaching, research, and scholarship activities, I have mastered excellent communication, organizational, and interpersonal skills.
Abstract:
Breast cancer brain metastases (BBM) represent a significant clinical challenge, with patients having a dismal 20% one-year survival. In the present study, we investigated the differential gene expression profile between tumor and normal samples by querying various public breast cancer cohorts, including TCGA BRCA, METABRIC, GSE11121, GSE17705, GSE12276, and CCLE. We compared the mean threshold in the breast cancer population, specifically in the ER+/HER2+ and TNBC, the more aggressive breast cancer subtypes. We further compared the expression levels across PAM50 subtypes to obtain unambiguous conclusions. The Kaplan-Meier (KM) analysis was performed to correlate overall survival curves based on gene expression data using the median as a threshold from TCGA and METABRIC breast carcinoma cohorts. Importantly, analysis done using TCGA-BRCA revealed elevated CBX2 mRNAseq gene expression in breast tumors than in normal tissue. Additionally, CBX2 mRNA seq expression showed a pattern of TNBC > HER2+ >all tumor >ER+ > normal tissue. Using Novartis/Broad Cancer Cell Line Encyclopedia (CCLE) for microarray and RNAseq gene expression profiling, we identified enhanced CBX2 mRNA expression in breast cancer cell lines compared to other cancer types. CBX2 mRNAseq expression was significantly greater among HER2+ (n=164) compared to normal (n=112) patients (FC=4.9; p=8.19e-36) and TNBC (n=115; FC=11, p-value= 1.27e-6) from TCGA breast cancer cohort. Additionally, METABRIC cohorts also showed a significant correlation between higher CBX2 mRNA expression and poor overall survival outcome (Kaplan-Meier analysis, n=1904, p- value= 0.0001). Thus, our investigation has identified CBX2 as a potential driver of HER2+ breast-to-brain metastasis and inhibition of CBX2 should be considered as a therapeutic target in breast-to-brain metastasis.
Biography:
Yan (Amber) Zhao has her expertise in molecular biology, microbiology, and cell biology. She has experience in process and assay development. Current research focuses on assay development to provide accurate and high quality IVD products for cancer diagnostic and monitoring.
Abstract:
Statement of the Problem: The nucleophosmin (NPM1) is the most mutated gene (~30%) in Acute Myeloid Leukemia (AML)1. Three NPM1 mutations (type A, B, and D) represent ~84% in NPM1-mutated AML cases while other uncommon subtypes occupy ~16%2. Xpert® NPM1 mutation, an automated cartridge-based test for measuring NPM1 mutation transcript levels (type A, B and D), is standardized to quantify the amount of mutated NPM1 relative to ABL1 control gene based on delta Ct in peripheral blood3. Since mutated NPM1 level is crucial for risk assessment, medication selection, and ongoing therapeutic monitoring in AML4,5, it is important to obtain the NPM1 mutation copy number (CN). The aim of this work is to develop NPM1 mutation CN estimator and to compare %NPM1 mutation/ABL1 reporting between delta Ct-based and CN-based methods. Methodology & Theoretical Orientation: Five levels of NPM1 mutations (A, B, D) and ABL1 IVT-RNA panels as well as two lots of Xpert® NPM1 mutation tests were used to generate standard curves for CN and %CN reporting. The cell lysates from cell lines carrying either NPM1 mutation A, B, or D and AML clinical samples containing NPM1 mutations were examined to evaluate the CN and %CN between two lots of the Xpert® NPM1 mutation tests and to compare the delta Ct-based and CN-based methods for reporting %NPM1 mutation/ABL. Findings: Linearity was demonstrated in Ct vs CN input for NPM1 mutation and ABL1 with R2 above 0.96 for Lot1 and Lo2. Less than 3-fold difference was exhibited for CN and %CN across two lots of Xpert® NPM1 mutation test. Less than 3-fold difference was observed in %NPM1 mutation/ABL1 reporting between delta Ct-based and CN-based approaches. Conclusion & Significance: An NPM1 mutation copy number estimator for Xpert® NPM1 mutation test was established, which will provide diagnostic and prognostic values for NPM1-mutated AML patients.
Biography:
Mengying(Mona) Liu has her expertise in molecular biology, microbiology, cell biology, gene & cell therapy, drug development, and biomarker discovery. She has experience in process and assay development. Current research focuses on assay development to provide accurate and high quality IVD products for cancer diagnostic and monitoring.
Abstract:
Statement of the Problem: Acute Promyelocytic Leukemia (APL) represents 10-15% of Acute Myeloid Leukemia (AML)1. The PML-RARa fusion transcript is expressed in more than 95% of APL patients2. Three PML-RARa isoforms (bcr1, bcr2, and bcr3) are identified in 90-95% of PML-RARa positive cases3. Prototype Xpert® PML-RARa, an automated cartridge-based assay for measuring PML-RARa fusion transcript levels (bcr1, bcr2, and bcr3), is standardized to quantify the amount of PML-RARA relative to ABL1 control gene based on delta Ct in peripheral blood. Since PML-RARa level is crucial for diagnosis and ongoing therapeutic monitoring in APL4,5, it can be useful to obtain the PML-RARa copy number (CN). The aim of this work is to develop PML-RARa CN estimator and to compare %PML-RARa/ABL1 reporting between delta Ct-based and CN-based methods. Methodology & Theoretical Orientation: Four levels of PML-RARa (bcr1, bcr2, and bcr3) and ABL1 IVT-RNA panels as well as two lots of prototype Xpert® PML-RARa assay were used to generate standard curves for CN and %CN reporting. The samples with spiked-in bcr1 IVT-RNA and APL clinical samples containing PML-RARa fusion transcript were examined to evaluate the CN and %CN between two lots of the prototype Xpert® PML-RARa assay and to compare the delta Ct-based and CN-based methods for reporting %PML-RARa/ABL1. Linearity was demonstrated in Ct vs CN input for PML-RARa (R2>0.98) and ABL1 (R2>0.97). Less than 2-fold difference was exhibited for CN and %CN across two different lots. Less than 2-fold difference was observed in %PML-RARa/ABL1 reporting between delta Ct-based and CN-based approaches. Conclusion & Significance: A PML-RARa copy number estimator for prototype Xpert® PML-RARa assay was established, which will provide helpful information for diagnosis and prognosis of APL. (Product in development. Not for use in diagnostic procedures. Not reviewed by any regulatory body.)