Slide 1: Breakthrough Cancer Detection Method
Breakthrough Cancer Detection Method Property of Volition© 2023 Slide 1
Slide 2: Forward-looking Statements and Disclaimer
Forward-looking Statements and Disclaimer Statements in this document may be “forward-looking statements” within the meaning of Section 27A of the Securities Act of 1933, as amended, and Section 21E of the Securities Exchange Act of 1934, as amended, that concern matters that involve risks and uncer
Slide 3: Our mission is to save lives and improve outcomes
Our mission is to save lives and improve outcomes for millions of people and animals worldwide. Property of Volition© 2023 Slide 3
Slide 4: Looking for cancer in a liquid biopsy is like a needle in a haystack
Looking for cancer in a liquid biopsy is like looking for a needle in a haystack ...and the But we found the needle is way to find it made of hay! Property of Volition© 2023 Slide 4
Slide 5: The Potential Opportunity: It's BIG!
The Potential Opportunity It’s BIG! $Billions$ Property of Volition© 2023 Slide 5
Slide 6: The Commercial Strategy: Licensing
The Commercial Strategy Licensing Property of Volition© 2023 Slide 6
Slide 7: We will seek upfront payments, milestones, royalties
We will seek • Upfront payments • Milestone payments • Royalties and • Sales of Key Components Property of Volition© 2023 Slide 7
Slide 8: Early cancer detection by plasma CTCF transcription factor analysis
Early cancer detection by plasma CTCF transcription factor analysis Dr Jake Micallef Chief Scientific Officer Property of Volition© 2023 Slide 8
Slide 9: Background
Background Property of Volition© 2023 Slide 9
Slide 10: Cancer and cell death cause chromatin fragmentation
Chromatin Chromosome Nucleosome Cancer and cell death cause chromatin CTCF fragmentation Sequence only present on CTCF in cancer Property of Volition© 2023 Slide 10
Slide 11: Liquid biopsy for ctDNA
Liquid biopsy for ctDNA Is like looking for a needle in a haystack… 99.9% 0.1% ...and the needle is made of hay! Nucleosomes and ctDNA released into the blood Property of Volition© 2023 Slide 11
Slide 12: Liquid biopsy for ctDNA
Liquid biopsy for ctDNA Wet chemistry pathway Amplification DNA Sample & library NGS Bioinformatics extraction preparation Methylated Septin-9 PCR Method unchanged since liquid biopsy first conceived What can you do with tens of millions of DNA sequences – other than sequence them? Property of Volit
Slide 13: Liquid biopsy for ctDNA
Liquid biopsy for ctDNA circulating tumor chromosome fragments DNA, DNA, DNA, DNA, DNA, DNA, DNA, DNA, DNA, DNA…, But DNA is DNA – you can’t separate out the bit you want! CANCER Property of Volition© 2023 Slide 13
Slide 14: Liquid biopsy for ctDNA… the real answer is KISS
Liquid biopsy for ctDNA… …the real answer is KISS 0.1% 99.9% Property of Volition© 2023 Slide 14
Slide 15: Liquid biopsy for ctDNA
Liquid biopsy for ctDNA CTCF 99.9% 0.1% Then you can do a You need to physically separate PCR test as simple some or all of the ctDNA as a COVID-19 test (nearly) Property of Volition© 2023 Slide 15
Slide 16: Novel wet chemistry pathway for ctDNA analysis
Novel wet chemistry pathway for ctDNA analysis NEW wet chemistry pathway PCR of CTCF Sample sequences ChIP collection uniquely present (Capture) in cancer First novel wet chemistry pathway for liquid biopsy First report of plasma TF-ChIP (Capture) Property of Volition© 2023 Slide 16
Slide 17: CTCF binding site occupation as epigenomic signature of cancer
Alterations to the binding site occupation pattern of CTCF is a functional epigenomic signature of cancer including both gain and loss of CTCF occupied sites in cancer cells.1 1. Fang et al. Genome Biology (2020) 21:247 Property of Volition© 2023 Slide 17
Slide 18: Hypothesis
Hypothesis = CTCF binding site with gain of occupancy in cancer CTCF Nucleosome Property of Volition© 2023 Slide 18
Slide 19: Hypothesis
Hypothesis Sequence present on nucleosomes Sequence is present in nucleosomes in healthy and in CTCF cancer CTCF Nucleosome Property of Volition© 2023 Slide 19
Slide 20: Hypothesis
Hypothesis Physically separate the CTCF from all the nucleosomes CTCF Property of Volition© 2023 Slide 20
Slide 21: Hypothesis
Hypothesis Extract the DNA CTCF Property of Volition© 2023 Slide 21
Slide 22: Hypothesis
Hypothesis Then cfDNA fragment with that sequence tumour derived CANCER CANCER CANCER Property of Volition© 2023 Slide 22
Slide 23: Hypothesis: ctDNA
Hypothesis ctDNA ctDNA ctDNA ctDNA Property of Volition© 2023 Slide 23
Slide 24: Method: CTCF-IP / qPCR Liquid Biopsy
Method: CTCF-IP / qPCR Liquid Biopsy CTCF Blood sample Nucleosome Wash DNA extraction qPCR Patient Test Result Plasma IP CTCF-occupied Short DNA Fragment cfDNA sequence Monoclonal <100 bp present/absent anti-CTCF Magnetic bead This method for the first time completely physically isolates ctDNA from
Slide 25: Biomarker Discovery
Biomarker Discovery CANCER Finding CTCF Binding sites occupied in cancer ctDNA – nucleosome covered in healthy and inflammatory plasma cfDNA Property of Volition© 2023 Slide 25
Slide 26: Method: Discovery
Method: Discovery Blood CTCF Healthy + sample Nucleosome inflammatory Wash DNA extraction DNA analysis NGS conditions Plasma IP Short DNA Fragment Monoclonal CANCER <100 bp anti-CTCF Develop qPCR Magnetic bead Property of Volition© 2023 Slide 26
Slide 27: Hypothesis in action
Hypothesis in action Property of Volition© 2023 Slide 27
Slide 28: Hypothesis in action
Hypothesis in action = CTCF binding site with gain of occupancy in cancer Healthy Cancer CTCF CTCF Nucleosome Nucleosome Property of Volition© 2023 Slide 28
Slide 29: Hypothesis in action
Hypothesis in action Sequence present on nucleosomes Sequence is present in in healthy nucleosomes and in CTCF cancer Healthy Cancer CTCF CTCF Nucleosome Nucleosome Property of Volition© 2023 Slide 29
Slide 30: Hypothesis in action
Hypothesis in action Physically separate the CTCF from all the nucleosomes Healthy Cancer CTCF CTCF Property of Volition© 2023 Slide 30
Slide 31: Hypothesis in action
Hypothesis in action Then cfDNA fragment with that sequence tumour derived Healthy Cancer CANCER CANCER CANCER Property of Volition© 2023 Slide 31
Slide 32: Hypothesis in action
Hypothesis in action ctDNA Healthy Cancer ctDNA ctDNA ctDNA Property of Volition© 2023 Slide 32
Slide 33: Clinical Experiment Results
Clinical Experiment Results Property of Volition© 2023 Slide 33
Slide 34: qPCR assays using CTCF-cfDNA biomarkers effective for leukemia detection
All 10 qPCR assays using CTCF-cfDNA biomarkers discovered for leukemia were effective for detection of leukemia • Some biomarkers were quite specific for leukemia • Most biomarkers were also effective for discovery of a variety of common solid cancers. • Different biomarkers were effective for diffe
Slide 35: Single qPCR assay detected 61% of leukemia cases at 98% specificity
A single qPCR assay discovered on leukemia detected Healthy Cancer 61% of leukemia cases at 98% specificity 1 1 1 0.249 1 1 1 1 0.28 352.139 1 137.663 1 35.383 3.89 4.155 1 1 6.17 41.643 ROC curve: Leukaemia (single qPCR assay) 2.85 30.274 6.39 20.252 0.83 4.857 100 0.66 11.314 5.37 1.58 1.439 1 80
Slide 36: Second qPCR detected 74% of leukemia cases at 96% specificity
A single qPCR assay discovered on Leukemia detected Healthy Cancer 61% of leukaemia cases at 98% specificity 1 1 1 0.249 1 1 1 1 0.28 352.139 1 137.663 1 35.383 3.89 4.155 ROC curve: Leukaemia (single qPCR assay) 1 6.17 1 41.643 2.85 30.274 100 6.39 0.83 20.252 4.857 0.66 11.314 80 . 5.37 1.439 1 1.
Slide 37: 2-qPCR assay detected 77% of colorectal cancer cases
Another 2-qPCR assay detected 77% of colorectal cancer cases at 92% specificity ROC curve: Leukaemia (single qPCR assay) Blood test almost as good as fecal 100 FIT test for colorectal cancer with biomarkers developed on 80 . leukemia. Sensitivity% 60 We are now developing individual 40 biomarkers sp
Slide 38: Proof of principle for multi-cancer early detection
Proof of principle for multi-cancer early detection using a 2-qPCR assay (90% specificity) Solid cancers patients positive sensitivity • CRC 13 9 69% • Breast 10 5 50% • Prostate 10 5 50% • Liver 10 6 60% TOTAL 43 25 58% Solid cancers patients positive sensitivity • Stage I 9 4 44% • Stage II 9 3 33
Slide 39: Comparison ChIP/PCR with traditional NGS/ctDNA assays
Comparison ChIP/PCR with traditional NGS/ ctDNA assays Traditional NGS/ ctDNA assay ChIP/PCR • Cost >$1000 • ~$100 • Too complex to be fully automated • Can be automated • High technology labs only • Any hospital lab • Turnaround - days or weeks • Turnaround 1 day (less if automated) • Library prepa
Slide 40: Dr Andrew Retter
Dr Andrew Retter Hematologist, Intensive Care, ECMO & Thrombosis Consultant, Guys and St Thomas’ NHS Foundation Trust, UK. Property of Volition© 2023 Slide 40
Slide 41: Conflicts of interest to declare
Conflicts of interest to declare Consultant to VolitionRX Limited No conflicts of interests. But just to confirm that these are my own opinions, and I am speaking in an independent capacity from the NHS Trust where I work as a consultant in hematology and intensive care Property of Volition© 2023 Sl
Slide 42: Cancer is the second leading cause of death worldwide
Cancer is the second leading cause of death worldwide. Property Volition Confidential of Volition© 2023 © 2022 Slide 42
Slide 43: Cancer Burden – new cases
Cancer Burden – new cases Property of Volition© 2023 Slide 43
Slide 44: Cancer Burden – death rate
Cancer Burden – death rate Property of Volition© 2023 Slide 44
Slide 45: Some further facts and figures…
Some further facts and figures… • Five-year survival rates are improving (from 49% in the mid-1970’s to 68% today) • Stage at diagnosis of cancer is an important factor that affects cancer outcomes. • Earlier diagnosis, when cancers are diagnosed at stages I and II as opposed to stages III and IV, i
Slide 46: How might Volition's technology help?
How might Volition’s technology help? • Diagnostic enrichment – early-stage diagnosis • Monitoring of disease progression and response to treatment …if we can diagnose earlier and monitor more efficiently outcomes should improve Property of Volition© 2023 Slide 46
Slide 47: What's next?
What’s next? • Clinical Paper submission, peer review and publication • Development of specific biomarkers for Lung, CRC, Prostate, Breast and Liver cancers • Cancer specific abstracts and posters at conferences throughout 2024 • Development of Centers of Excellence and Key Opinion Leaders Property
Slide 48: In Summary
Updates In Summary • Exciting breakthrough method • A number of “firsts” • Significant Clinical and Commercial opportunity Property of Volition© 2023 Slide 48
Slide 49: Question & Answer Session
Question & Answer Session Property Volition Confidential of Volition© 2023 © 2022 Slide 49