This document discusses an approach to detecting and treating epigenetic marks in circulating tumor cells (CTCs) across solid tumors. It summarizes a three-year study collaborating between Sahel Oncology and Orange Coast Medical Center of Hope that tested for HDAC1 and DNMT1 expression in CTCs of 45 cancer patients. They found elevated HDAC1 or DNMT1 in 47-55% of cases. These patients were treated with an epigenetic therapy called MTET targeting HDACs and DNMTs. MTET was effective in 40% of patients as shown by reductions in CTCs two weeks post-treatment. There was an overall 70% response rate seen on imaging for cases with positive CTCs.

1. DETECTION AND TREATMENT
APPROACH TO EPIGENETIC MARKS IN
THE CTC ACROSS SOLID TUMORS
M.NEZAMI MD
SAHEL ONCOLOGY LLC-WWW.SAHELONCOLOGY.COM
ORANGE COAST MEDICAL CENTER OF HOPE-WWW.ALLCANCERCARE.COM

2. SAHEL ONCOLOGY
• Early stage drug development company with significant IP profile and data ( invivo)
• Phase I and II trials, Approved orphan drug designation( ODD)
• Technology: MTET( MultiTargeted EpigeneticTherapy). Combination of HDACi and
DNMTi ( Qc- NP/PGLA)
• www.saheloncology.com
• M. Nezami MD, President

3. AGENDA
• Learning history of circulating tumor cell analysis in therapeutic approaches
• Identification of predictive epigenetic marks in CTC
• Understanding treatment design in a case series of solid tumors and clinical
outcomes

4. CTC
• CTCs are prognostic both on detection and quantification. ( Patient survival will be
directly impacted by the presence or absence of the CTC). The mRNA analysis of CTC
has created a platform for therapeutics.
• The actual understanding of CTC epigenome will create a novel approach as companion
diagnostics for epigenetic therapies.
• Multitargeted epigenetic therapy( MTET) is a patented therapy to target HDAC and
DNMT in CTC, and has show superior results in inhibition of epidermo-mesenchymal
transition ( EMT) and as such to reduce the tumor ability to disseminate.

8. BIOFOCUS LAB

9. BIO FOCUS LABORATORY ( TESTING CTC)
• Methods
• Gene expression analysis of HDAC1 and DNMT1 in CTCs was performed by realtime RT-PCR. Primers
and Taqman probes were designed by using Realtime PCR Tool on the Integrated DNA Technologies Inc.
(IDT) website on refseq sequences for HDAC1 (NM_004964) and DNMT1 (NM_001130823,
NM_001379). Care was taken that primers and/or probes are spanning exon-exon boundaries.
• Gene expression was measured in CTCs and corresponding normal mononuclear blood cells,
normalized to house-keeping gene GAPDH.
• A greater than two-fold relative expression in CTCs to normal mononuclear blood cells was considered
elevated.

10. STUDY
• Three years ( 2020-2021-2022) of collaboration between Sahel Oncology with a
designated cancer center called Orange Coast Medical Center of Hope in California, and
Biofocus Lab; Biofocus LADR Gesellschaft für biologische Analytik mbH
• 47-55 percent positive HDAC OR DNMT tested in total of 45 cases. The combination
of alterations were seen in 18 to 25 percent of case.
• These cases were treated with epigenetic therapies targeting these findings and results
reported here for the first time.

11. RESULTS
Year patients(n)
HDAC1 % DNMT1 % HDAC1 or DNMT1 % HDAC1 and DNMT1 %
2020 20 11 55 5 25 11 55 5 25
2021 8 3 38 3 38 4 50 2 25
2022 17 6 35 5 29 8 47 3 18
all 45 20 44 13 29 23 51 10 22
elevated (n / %)

12. CTC RESPONSE TO MTET

13. CONCLUSION
• MTET was effective in 40 percent of patients samples evident in their sequential post
therapy analysis 2 weeks after therapy. 50 percent of samples had complete eradication
of CTC and 50 percent had partial reduction.The clinical outcome in our case study
correlated with liquid biopsy findings. In cases with positive CTC, there was an overall
response of 70 percent evident by imaging.
• MTET can be considered as an effective method of treatment alone or in combination
with other modalities for solid tumors with positive CTC with epigenetic marks. Further
clinical trials are suggested to assess the long term survival and correlation with
eradication of CTC.

14. THANKYOU
• www.saheloncology.com
• www.allcancercare.com
• 949-515-4673
• Questions?