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AJS: a lethal weapon to combat with cancers Treatment of  Human Cancers Using Mechanism-Based Lentivirus
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AJS: a lethal weapon to combat with cancers Treatment of Human Cancers Using Mechanism-Based Lentivirus

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Cdc6 is essential in the initiation of DNA replication: …

Cdc6 is essential in the initiation of DNA replication:
A). Assembling pre-replication complex (pre-RC) with ORC, and loading MCMs to replication origins;
B). Licensing DNA replication to ensure one round DNA replication per cell cycle.

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  • 1. AJS: a lethal weapon to combat with cancersTreatment of Human Cancers Using Mechanism-Based Lentivirus Luo Feng Takeshi Hirota Tsuyoshi Takara RNtein Biotech Co., LtD Tokyo, Japan RNTein Biotech. Lab. Dedicated to human cancer gene therapy San Gabriel, California, USA
  • 2. Self-sufficiency Sustained in growth signals angiogenesis Insensitivity to anti-growth signalsLimitless replicative Evading apoptosis Tissue invasion potential & metastasis The development of human cancer cells
  • 3. Cdc6 is essential in the initiation of DNA replication: A). Assembling pre-replication complex (pre-RC) with ORC, and loading MCMs to replication origins; B). Licensing DNA replication to ensure one round DNA replication per cell cycle.
  • 4. The novel function of AJSFL1 is the silencing of human Cdc6, the specific genes for cell proliferation: Specific Cdc6 shRNAs produced from the viral vector Inhibit Cdc6 and block Cancer cell proliferation, but not normal cells. LA-N-2 MCF7 CHLA255 Cdc6 shRNA THTD6Ai THTD6Ai THTD6Ai THTP Mock Mock THTP THTP Cdc6Cdc6 knockdown -actin 1 2 3 4 5 6 7 8
  • 5. MRecombinant G1 Lentivirus AJS2001 G2 STOP S
  • 6. Cdc6 Telomere Oncogene dysfunction activation ? Rb-E2F Cyclin D1 Cdk4/Cdk6 DNA damage p16INK4a P Rb-E2F response p53 p21 P Rb E2F Rb Apoptosis Senescence G1 SAdapted from J. M. Sedivy, Cancer Cell, 11:389-391, 2007
  • 7. Reactivation of p16INK4a CDK inhibitory activity by Cdc6 knockdown — —    THTD — — — IP/mouse —IgG    IP/β- p16 INK4a Phosph-Rb-C Ser780 Phosph-Rb-C Ser807/811 IgG, light chain 1 2 3 4 — — —  THTD 1:10 1:25 1:5  IP/β- p16 INK4a Phosph Rb-C Ser 807/811 IgG, light chain 1 2 3 4
  • 8. Senescence-associated β-glactosidase (SA-β-Gla) positive staining caused by Cdc6 knockdown MCF7 MCF7/THTP MCF7/THTD6Ai
  • 9. The formation of senescence associated heterochromatic foci via Cdc6 knockdown Anti-H3K9me3/me2 indirect fluorescent staining Mock THTD6Ai Confocal fluorescent microscopy Mock THTD6Ai
  • 10. A male patient age 55, with advanced gastric carcinoma found in April 2006 is first survivor byGFL1(AJS). After second injection since 07/06/2006, he gained weight with good appetite, and lefthospital on 27/06/2006. After 4th injection, his gastric carcinoma disappeared without side effect.Now, 2 years and 8 month has passed since recovery, and he leads normal life with good appetite  and feeling. He and his wife filled with joy • • • has become active for social workSpeech about his own recovery experience at Cancer Control Association. He is now doingvolunteer activity for many other cancer patient. An advanced gastric carcinoma patient shows his recovery after receiving AJS injections.
  • 11. Multiple gastric scirrhous carcinoma of the male patient age 55 Emergency medical admission on 10/04/2006  First injection on 24/04/2006 2 weeks after 4 times of weekly injection on 05/06/2006 Effect of AJS treatment shown by endoscope
  • 12. A female patient age 52, who had major operation for uterus carcinoma metastated to liver and brainresulted in 14 days life expectancy because brain carcinoma remain, has amazingly recovered bycontinuous AJS injection for 3 month. Now, she leads pleasant normal life CT of brain before injection CT of brain after recovery MRI of liver before injection Healthy expression after recovery The treatment of a patient with brain metastatic ovary tumour
  • 13. A male patient age 64, recovered from parotid carcinoma metastated to lung and larynx. Hehad the therapy by day return to keep his office work. He has been nominated to chairman ofI (AJS) patient association. He has now 6 grandchild, and is doing volunteer activity topromote AJS. Before injection After injection
  • 14. A female patient age 52, has recovered from breast cancer metastated to lymph node by only 4 times injection without surgery. In addition, her hair returned deep black. She is now doing volunteer activity for many other breast cancer patient by saying “do not need surgery, do not give up hope, breast cancer can be cured by injection only”. Before injection After 1st injection After 2nd injection After 3rd injection After 4th injection Current recovery Her black hairA breast carcinoma patient voluntarily wants to have AJS treatment, not surgery.

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