Thor Nilsen NeoGeneStar LLC January 22, 2015
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1 Thor Nilsen NeoGeneStar LLC January 22, 2015
2 NeoGeneStar TM I. Liquid biopsy using circulating cell-free DNA: Cancer diagnostics Prenatal diagnostics Other disease states II. NeoGeneStar TM cell-free DNA purification kits
3 NeoGeneStar In the United States: Every 18 seconds, a person learns that he/she has cancer, he/she want to know Every 8 seconds, a baby is conceived, every parent wants to know
4 Pregnant? cancer? other diseases? No more tissue biopsy! Introducing liquid biopsy Simple and non-invasive Draw blood or collect urine Purify circulating cell-free DNA (cfdna) Analyze cfdna (NGS, PCR ) Get results quickly
5 cfdna serves as a biomarker in clinical diagnosis: cfdna: DNA not associated with cells Apoptosis related, multiples of 180bp, 180bp, 360bp, 540bp Necrotic related: 1~10kb Microvesicles: mostly ssdna Exosomal: mostly dsdna Pathogenic: cfdna in normal persons: about ng/ml Cancer patients: significantly increased >100ng/ml cfdna is used as a biomarker in early detection, and monitoring of the therapeutic response of cancer treatment regimens. cfdna is also a useful biomarker in other disease states, such as stroke, embolism, cardiac ischemia, trauma and autoimmune diseases
6 Circulating Cell free Tumor DNA in Cancer Detection and Monitoring Personalized Cancer Therapy Tracking Cancer Resistance Cell-free Tumor DNA Early Cancer Detection Monitoring Tumor Burden
7 Molecular Profile of Tumor Heterogeneity to guide Therapy Liquid Biopsies to Monitor Response and Resistance to Targeted Therapies Luis Alberto Diaz, M.D., Associate Professor of Oncology, Johns Hopkins Sidney Kimmel Comprehensive Cancer Center Application of Clinical Tumor Genotyping in Targeted Cancer Therapy Darrell R. Borger, Ph.D., Co-Director, Translational Research Laboratory, Massachusetts General Hospital Cancer Center Quantitative Tumor Protein Profiling for Therapy-Relevant Stratification of Breast Cancer Patients Hallgeir Rui, M.D., Ph.D., Professor, Cancer Biology, Medical Oncology and Pathology; Scientific Director, Jefferson Breast Care Center; Program Leader, Biology of Breast Cancer, Kimmel Cancer Center; Co-Director, Pathology Translational Research Core, Thomas Jefferson University Clinical Validation of Predictive Biomarkers and Next-Generation Personalized Medicine Treatment Strategies Incorporating Genetic Dynamics Robert A. Beckman, M.D., External Faculty, Center for Evolution and Cancer, Helen Diller Family Cancer Center, University of California at San Francisco; Executive Director, Clinical Development Oncology, Daiichi Sankyo Pharma Development
8 Diehl et al Nature Medicine, 2008
9 Correlation of cfdna with surgeries and cancer progression Chemotherapy Sigmoid colectomy + left lateral hepatic resection + right liver metastasis left Chemotherapy in place SURGERY SURGERY Right hepatectomy Total DNA fragments in 2 ml of plasma (n) Total DNA (PIK3CA) Total DNA (TP53) Time (days) Data cited from Dr. Luis Diaz
10 Purification of Cell-free DNA for Cancer Early Detection & Monitoring: Liquid Biopsy: blood or urine test, non-invasive When the tumor is not accessible: cell-free DNA is the best analyte available Especially useful following surgery and/or chemotherapy: Recurrence can be detected using cell-free DNA in blood No detection at 6 months & 1 year is great news
11 Diehl et al, Nature Medicine, 2008 No Detectable Cancer DNA = 100% Survival Detection of ctdna is predictor of poor outcome
12 Summary: Application of cfdna in Cancer Diagnostic Therapeutic Early Detection Liquid biopsy (genotyping) Tumor dynamics Monitor the evolution of resistance in realtime Assess minimal residual disease Assess heterogeneity Identify targets for drug creation Assess early treatment response or failure Informed drug development Hypothesis testing of pathway inhibition
13 Circulating Cell-free Fetal DNA
14 Discovery of Cell-free Fetal DNA in Maternal Blood Dr. Dennis Lo: Y chromosome detected in maternal blood (1997) Cell-free fetal DNA is present in maternal circulation Non-invasive prenatal tests (NIPT) No risk of miscarriage, infection or injury Detectable as early as 5 weeks of gestation ACOG recommends that prenatal screening using cffdna be available to all pregnant women (2012) Major healthcare companies accept NIPT as medically necessary for high risk pregnancies (2013) ACOG is American Congress of Obstetricians and Gynecologists
15 Clinical Prenatal Diagnostic Options: Invasive Tests: Amniocentesis - risk to fetus Chorionic villus sampling - risk to fetus Embryoscopy and fetoscopy - risk to fetus Percutaneous umbilical cord blood sampling - risk to fetus Non-invasive tests: Blood tests Triple test / Quad test - limited information Ultrasound - limited information Non-Invasive Prenatal Tests (NIPT): Cell-free fetal DNA in maternal blood Early detection: 8-10 weeks of gestation provides fetal genetic information
16 Detectable earlier during pregnancy (usually 8-10 weeks) Non-invasive, no risk of miscarriage, infection or injury Accuracy (using PCR or Next Generation Sequencing) Downs, Edwards and Patau Syndromes (Trisomies) Sex chromosome aneuploides (abnormal number of sex chromosomes) Single gene associated diseases such as cystic fibrosis The earlier an expectant mother get results, the earlier she can make an informed decision
17 NeoGeneStar TM Cell-free DNA Purification Kits Patent pending technology Magnetic bead capture: no filtration, no clogging, no need for centrifugation Expect near 100% recovery of cell-free DNA from plasma or urine Highest yield of purified cell-free DNA Special designed magnetic stands to fit with existing robots for automation
18 Purification of circulating cell free DNA using NeoGeneStar cell free DNA purification kit Cutting edge superparamagnetic nanotechnology Highly effective method for cfdna purification Simple procedures ~45 minutes No specialized equipment required No vacuum or centrifugation Automatable
19
20 Data courtesy of Jessica Padilla, Cornell University
21 Data courtesy of Arup Laboratories
22 180bp cfdna Control Purified cfdna Urine Total DNA
23 Just Magnets and Pipets are needed for the NeoGeneStar TM cell-free circulating DNA kits NeoGeneStar 8 & 16 position magnetic stands 96 well (standard & deep) 24 well - deep to 10ml volume
24 Key advantages of NeoGeneStar TM products: Highly effective method for cfdna purification Applicable for isolation of cfdna from cancer patients Applicable for screening, early detection and monitoring therapeutic progress of cancer patients Amenable to automation for large numbers of samples Cost effective / profitable at competitive pricing cffdna is cell-free fetal DNA in maternal peripheral blood cfdna cell-free DNA from cancer patients and other disease states
25 Nancy Quan, MD, Chief Executive Officer Phone number: address: Thor W Nilsen, VP, Business Development Phone number: address: ThorWNilsen@NeoGeneStar.com
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