Tumor monitoring in a drop of blood.

Transcription

1 Tumor monitoring in a drop of blood. PRECISION MEDICINE STARTS WITH A FINGER STICK. circulogene.com

2 Circulogene Theranostics solution to liquid biopsy. Novel cell-free DNA enrichment method for high quantity and high quality starting materials: Circulating, cell-free DNA is highly fragmented and presented at very low concentrations; therefore, its isolation is challenging. It requires high volume input, costs, and labor intensity. Circulogene s proprietary, cell-free DNA enrichment process possesses novel characteristics of low input (as low as 50 ul of blood) and high output (>300ng/mL), allowing ultra-high recovery and detection of tumor DNA directly from a droplet volume of unprocessed plasma. Advanced NGS technology for sensitive longitudinal mutation detection: The updated EURTAC study demonstrates that mutations detected in cfdna are prognostic and consistent with data obtained from tumor biopsies, the editorialists wrote. More broadly, the potential benefits of liquid biopsies include a better evaluation of the tumor genome landscape with the identification of a comprehensive set of targetable mutations and the serial noninvasive monitoring, which may allow the detection of additional mutations from emerging subclones, including those involved in the development of acquired resistance. Detection of multiple, low-frequency mutations through periodic cell-free DNA analysis could predict tumor progression before the lesions are large enough to be detected by imaging. Analysis beyond a single mutation could also be used to identify and quantify tumor heterogeneity for effective treatment decision making. Liquid biopsies are not as spatially limited as tissue biopsies, and can show a global spectrum of mutations that occur throughout tumor development. The sensitivity of conventional analytical methods such as Sanger sequencing is not sufficient to detect low-frequency variants. Targeted deep sequencing by next-generation sequencing (NGS) provides a cost-effective alternative for high-throughput analysis of multiple mutations with high sensitivity. Validated sample processing and analysis procedure in clinical practice: The utility of cell-free DNA for clinical application demands the implementation of stringent pre-analytical validation of our laboratory-developed tests to ensure consistent quality data acquisition in our CLIA-certified laboratory.

3 Through our patented, liquid biopsy process, Circulogene Theranostics delivers tumor monitoring from droplets of blood. Non-Invasive Precision Medicine Because Circulogene s patented technique enriches the sample, analysis only requires 50 microliters of blood from a finger stick, which is more convenient for healthcare professionals and patients. Accurate Identification of Tumor Mutations Circulogene s tests analyze for the presence of cell-free DNA, identifying and quantifying any tumor mutations for accurate reporting and monitoring. The CORRECT trial was the first large clinical trial to compare liquid versus conventional tissue biopsy data, and the results show that liquid biopsy obtains more data on tumor mutations throughout the course of the disease, enabling us to better target therapy to the specificities of a patient s tumor, Josep Tabernero, M.D., Ph.D., M.Sc., head of the medical oncology department of Hospital Universitario Vall d Hebron and author of the CORRECT study. Tumor Mutations Matched with Treatment In our personalized gene reports, Circulogene further provides information on current FDA-approved treatment options proven effective for the tumor DNA mutations identified. Track Responsiveness Frequently The efficiency of Circulogene s process empowers physicians to closely monitor tumor responsiveness and adjust treatment protocols. Turnaround time is 5 to 7 days, much faster than tissue biopsy results and other liquid biopsies. Tumor Burden and Recurrence The Circulogene process uses precise baseline data from which to measure cell turnover and progression. A Yield of 1,000x with 1/10 of the Sample We compared the sensitivity of our proprietary cell-free DNA enrichment method to the standard DNA extraction kit used industry-wide today. Results demonstrated 1,000x more cell-free tumor DNA in the original sample using a single drop of blood.

4 Precision Medicine Technological advances combined with an improved understanding of the genetic basis of cancer has allowed individualized profiling that is revolutionizing personalized medicine and cancer treatment. With a simple finger stick, Circulogene s patented technology utilizes next-generation sequencing (NGS) to monitor known tumor mutations (approximately 3,000) in the scientifically established cancer-associated genes listed below. Circulo Breast ERBB2 Bose et al. Cancer Discov Feb;3(2): AKT1 Bleeker et al. Oncogene Sep 18;27(42): Lindsley CW. Curr Top Med Chem. 2010;10(4): PTEN Lauriol & Kontaridis. Trends Cardiovasc Med May; 21(4): Tartaglia M et al. Am J Hum Genet Feb; 78(2): Circulo Colorectal Circulo Lung Schubert et al. Nat Rev Cancer Apr;7(4): Romano et al. Clin Cancer Res Oct 15;19(20): EGFR Brooks et al. Clin Cancer Res Apr 1;18(7): ALK Choi et al. N Engl J Med Oct 28;363(18): Sasaki et al. Cancer Res Sep 15;71(18): Circulo Hematological Circulo Thyroid FLT3 Baker et al. Clin Cancer Res Oct 15;19(20): Smith et al. Nature Apr 15;485(7397): NPM1 Estey EH. Am J Hematol Apr;88(4): JAK2 Scott. Am J Hematol Aug; 86(8): Kiladjian et al. Blood. 2006;108: Scott et al. Blood ABL1 Santos et al. Cancer J Nov-Dec;17(6): Soverini et al. Blood Aug 4;118(5): RET Phay and Shaw. Clin Cancer Res Dec 15;16(24): Schubert et al. Nat Rev Cancer Apr;7(4): Romano et al. Clin Cancer Res Oct 15;19(20): Circulo Melanoma Note: Circulogene does not mandate or advise treatment for individual patients. Final recommendations are the responsibility of the clinician. Schubert et al. Nat Rev Cancer Apr;7(4): Romano et al. Clin Cancer Res Oct 15;19(20): KIT Heinrich MC et al. J Clin Oncol 2003;21(23): Debiec-Rychter et al. Eur J Cancer 2006;42(8): v

5 Circulo Ovarian PTEN Lauriol and Kontaridis. Trends Cardiovasc Med May;21(4): Tartaglia M et al. Am J Hum Genet Feb;78(2): Circulogene s results provide valuable and actionable information for physicians evaluating tumor resistance and recurrence, metastasis, prognosis, and treatment protocols and progress. Circulo Gastric APC Minde et al. Mol Cancer Aug 22; 10:101 Knudsen et al. Fam Cancer. 2003;2(1): Circulo Pancreatic Each personalized gene profile provides information on current FDA-approved treatment options proven effective for the tumor DNA mutations identified. ABL1 Bosutinib Busulfan Dasatinib Imatinib Nilotinib Omacetaxine Ponatinib AKT1 ALK Ceritinib Crizotinib FLT3 Cabozantinib Ponatinib JAK2 Ruxolitinib KIT Axitinib Cabozantinib Imatinib Pazopanib Regorafenib Sorafenib CDKN2A Goldstein AM et al. Med Genet. 2007; 44: Ghiorzo P et al. Ann Oncol Jan;15(1). TP53 Gonzalez et al. J Clin Oncol. 2009; 27(8): Levine. Cell Feb 7;88(3): Circulo GIST KIT Heinrich MC et al. J Clin Oncol 2003;21(23): Debiec-Rychter et al. Eur J Cancer 2006;42(8): PDGFRA Corless et al. J Clin Oncol Aug 10;23(23): Heinrich et al. Science Jan 31;299(5607): APC Dabrafenib Trametinib Vemurafenib CDKN2A EGFR Afatinib Cetuximab Erlotinib Gefitinib Lapatinib Panitumumab Vandetanib ERBB2 Ado-trastuzumab emtansine Everolimus Lapatinib Pertuzumab Trastuzumab Cetuximab Panitumumab NPM1 PDGFRA Axitinib Imatinib Pazopanib Sorafenib PTEN RET Cabozantinib Regorafenib Vandetanib TP53 Sources: cancer.gov and fda.gov

6 Personalized Gene Profile Sample Patient Name: Client Name: Lims ID: DoB: Age: Gender: Ordering Physician: Collection Date: Date Received: Indication: Colorectal Cancer Client ID: Date Reported: Sequence Variants: 1 FDA Biomarker Guidance MUTANT FDA GUIDANCE FDA GUIDANCE c.38g>a; p.g13d 21.0% Cetuximab Contraindicated TRIALS 22 ALTERATION FRACTION Panitumumab Contraindicated (for indication) (for other indication) (details below) c.34g>a; p.g12s 3.2% Cetuximab Contraindicated 22 c.35g>t; p.g12v Codon 12 Panitumumab Contraindicated 97.4% Y 23 c.34g>t; p.g12c 1.7% Cetuximab Contraindicated 22 Panitumumab Contraindicated *Variants are reported if sequence coverage is greater than 100x, quality is greater than 7, variant frequency is greater than c.3140a>g; p.h1047r 36.3% 4 c.1799t>g; p.v600g 1.8% 16 c.1799t>a; p.v600e 5.7% Melanoma (V600E): 16 Cetuximab Colorectal Cancer codon 12 and 13 mutation Dabrafenib, negative Trametinib & Vemurafenib Contraindicate LABEL INFORMATION Cetuximab is indicated for wild-type, EGFR-expressing, metastatic colorectal cancer as determined by FDA-approved tests. Cetuximab is not indicated for the treatment of patients with colorectal cancer that harbor somatic mutations in exon 2 *Variants are reported if sequence coverage is greater than 100x, quality is greater than 10, variant frequency is greater than 1%. (codons 12 and 13), exon 3 (codons 59 and 61), and exon 4 (codons 117 and 146) of either K-Ras or N-Ras. FDA Biomarker Guidance No Reported Mutation 8 Panitumumab Colorectal Cancer codon 12 and 13 mutation negative Contraindicate LABEL INFORMATION Panitumumab is indicated for the treatment of wild-type metastatic colorectal cancer as determined by an FDAapproved test. Panitumumab is not indicated for the treatment of patients with colorectal cancer that harbor somatic Panitumumab Colorectal Cancer codon 12 and 13 mutation negative mutations in exon 2 (codons 12 and 13), exon 3 (codons 59 and 61), and exon 4 (codons 117 and 146) of either or GUIDANCE Contraindicated Panitumumab is indicated for the treatment of wild-type metastatic colorectal cancer as determined by an FDA-approved test. Panitumumab is not indicated for the treatment of patients with colorectal cancer that harbor somatic mutations in exon 2 (codons 12 and 13), exon 3 (codons 59 and 61), and exon 4 (codons 117 and 146) of either or FDA BIOMARKER INFORMATION FDA BIOMARKER INFORMATION The Information compiled in this report The Information is from the compiled FDA's in Table this report of Pharmacogenomic is from the FDA's Table of Biomarkers Pharmacogenomic in Drug Biomarkers Labeling, Drug augmented Labeling, augmented with with information nformation extracted from the drug extracted label at from the the FDA. drug The label list at the does FDA. not The represent list does not a represent listing of a listing all drugs of all drugs that that might be be approved for for the the indication, but only ndication, but only those drugs approved those drugs for the approved specific for the biomarker, specific biomarker, indication indication and and referenced subgroup. Lab Director: Dr. Kathryn Clary Client Services: Fax: Independence Drive, Suite 301, Birmingham, AL support@circulogene.com Open Clinical Trials: 23 RESEARCH USE ONLY- Not for Diagnostic Purposes 1 NCT Phase 1/ NCT Phase 1/ A Phase I/II Dose-Escalation Study Evaluating the Combination of Neratinib and Cetuximab in Patients With "Quadruple Wild- Type" (/// Wild-Type) Metastatic Colorectal Cancer Resistant to Cetuximab Drug:Cetuximab; Drug:Neratinib; STATE(S) FL; ID; IL; MI; NC; PA Drug:WNT974; Drug:LGX818; Biological:Cetuximab; Colorectal Cancer; A Phase Ib/II Multi-center, Open Label, Dose Escalation Study of WNT974, LGX818 and Cetuximab in Patients With V600- mutant Wild-type Metastatic Colorectal Cancer Harboring Wnt Pathway Mutations STATE(S) FL; SC; TX; WI; MI Metastatic Colorectal Cancer;

7 Open Clinical Trials Continued NCT NCT Phase 1 NCT NCT NCT NCT A Randomized, Double-blind, Placebo-Controlled Phase II Trial of Irinotecan, Cetuximab, and Bevacizumab Compared With Irinotecan, Cetuximab, and Placebo in -Wildtype, Irinotecan-Refractory, Metastatic Colorectal Cancer Drug:Bevacizumab; Drug:Irinotecan; Drug:Cetuximab; STATE(S) IA; KS; LA; MA; MN; MO; NH; NY; WI Biological:Panitumumab; Drug:Cabozantinib; A Randomized Phase II Study of Irinotecan and Cetuximab With or Without the Anti-Angiogenic Antibody, Ramucirumab (IMC -1121B), in Advanced, K-ras Wild-Type Colorectal Cancer Following Progression on Bevacizumab-Containing Chemotherapy Biological:cetuximab; Biological:ramucirumab; Drug:irinotecan hydrochloride; Colorectal Cancer; STATE(S) AL; CO; CT; GA; IL; IN; IA; KS; LA; MD; MA; MI; MN; NE; NJ; NM; ND; OH; OK; PA; SD; TX; VA; WV; WI A Phase II Study of Panitumumab in Combination With FOLFIRI After Progression on FOLFIRI Plus Bevacizumab in (Kirsten Rat Sarcoma) and Wild-Type Metastatic Colorectal Cancer. Biological:panitumumab; Drug:irinotecan hydrochloride; Drug: fluorouracil; Drug:leucovorin calcium; STATE(S) OH; TN Metastatic Colorectal Cancer; Cabozantinib (XL184) With Panitumumab in Subjects With Wild-Type Metastatic Colorectal Cancer and Cabozantinib Monotherapy in Subjects With MET Amplified Treatment-Refractory Colorectal Cancer STATE(S) NC Drug:panitumumab; Drug:Randomization to No Panitumumab; Colorectal Cancer; A Randomized Phase II Study of Hepatic Arterial Infusion With Intravenous Irinotecan, 5FU and Leucovorin With or Without Panitumumab, in Patients With Wild Type Who Have Resected Hepatic Metastases From Colorectal Cancer STATE(S) NJ; NY Biological:BAX69 + infusional 5-FU/LV; Biological:BAX69 + panitumumab; Biological:BAX FU/LV; Biological:BAX69 + panitumumab; Drug:Standard of Care; Biological:Standard of Care; Metastatic Colorectal Cancer; A a Randomized, Open-label Study to Assess the Safety, Tolerability, and Efficacy of BAX69 in Combination With 5- FU/Leucovorin or Panitumumab Versus Standard of Care in Subjects With Metastatic Colorectal Cancer STATE(S) IL Metastatic Colorectal Cancer; Mucinous Adenocarcinoma of the Colon; Mucinous Adenocarcinoma of the Rectum; Recurrent Colon Cancer; Recurrent Rectal Cancer; Signet Ring Adenocarcinoma of the Colon; Signet Ring Adenocarcinoma of the Rectum; Stage IV Colon Cancer; Stage IV Rectal Cancer; Note: Circulogene does not mandate or advise treatment for individual patients. Final recommendations are the responsibility of the clinician.

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