PAPS - Rapamycin Induces Autophagy And Apoptosis In Kaposiform Hemangioendothelioma Primary Cells In Vitro - Zuopeng Wang

We have Dr. Zopang Wang from Hong Kong who will present on rapamycin inducing autophagy and apoptosis, um, in primary cells in vitro. And then following Dr. Yamataka from Jundo University School of Medicine in Tokyo will give his take on all of these results. Um and everyone else remember, absolutely post questions or comments in the chat. We're glad to uh read them out during the conversation and we'll make sure everyone has their minute in the ring. Good morning, I'm Zo Pang Wang from Children's Hospital of Fudan University, Shanghai, China. Today, my topic is, Rapamycin induces autophagy and apoptosis in cupusyform hemangioendothelioma primary cells in vitro. KHE is a rare and distinct vascular tumor affecting children with life threatened Kabasa Marat phenomenon, KMP. Rapamycin inhibits the MTOR pathway and has been reported to be safe and efficient in treating KHE with KMP. However, the lack of KH cell lines causes a trouble to carry out its mechanism studies. So we culture the KH primary cells from a one month old male infant diagnosed of KH with KMP. And the patient was confirmed with histopathology. The specimen were cut into tiny pieces and digested. Then the primary cells were selected by magnetic bead with CD34. Then we explore the cell proliferation, apoptosis, autophagy. The patient was diagnosed with KH by H standing, positive for CD31, CD34, LV1, D240 and prox1. After isolation, we can found that the KH primary cells is also spinal shaped. The immunofluorescence stands show that the primary cells was positive for CD31 and LYV1, but was negative for D240. Then we explored the cell proliferation and we found the cell viabilities was significantly inhibited in the repa group. And we also found rapamycin induced G0 and G1 phase rest resulted in high level of apoptosis. Then we detecting the autophagy and the mTOR pathway by western broading. And the ratio of LC32 and 1 increased in rapamycin glooper, symbolizing the occurrence of autophagy. And the level of fosforation and his proteins are significantly suppressed in rapamycin group. The immunofluorescence of LC3 also indicated that rapamycin induced autophagy of KH primary cells. So in conclusion, our study determined that rapamycin inhibited KH primary cells proliferation, induced G0 and G1 phase arrest, apoptosis, autophagy, and providing evidence that rapamycin could be a useful approach to treat KH. Thank you very much. Dr. Wang, thanks for your nice presentation. And uh it is a great study at clinical pathological uh, you know, including uh clinical and basic study and uh my question is uh uh did you investigate how the uh effects other cells other than the uh the uh you know, the cellss. I mean the other cells have a had a side effect in your study if you have checked. Yes, I have tried in the neuroblastoma and uh we also observed the similar uh effect such as uh it can inhibited the uh uh it can induced the apoptosis in the uh S the neuroblastoma cells. So I think it may be a uh a drug will use in the future for the tumors uh therapy. So have you checked the uh, have you applied your technique into the uh clinical cases? Um, we we use uh we we tried in some uh uh the stage four tumor patients uh but uh uh we we combined with the chemotherapy. So uh we cannot uh uh to to determined which uh has the effect uh uh as the dominant. So maybe we need the clinical child to found the effect in the future. I hope you will get the clinical clinical study application the kind of a pilot study. So, so I want to ask audience uh the uh questions comment or panelist. Do you have any comments or questions from panelist first? Any any comments from the analysts about this paper? So I just wanted to clarify one thing and then um you guys maybe I don't know John if it messes up the play if they raise their hand if they have a question but um so I just want to clarify the last thing that Yama just said. So is the next step for you now going to be clinical trials or is there another step in between that you have to do before you actually start clinical trials? Uh, maybe it it just uh uh we we we now we use the uh rapamycin to treat the KHE and uh we focus on KH therapy and uh we uh at present we just uh try the uh uh clinical trials for uh the rapamycin and the rapamycin with other drugs to treat the KHE. And for the tumor therapy maybe it will uh in the future not in the present. So my the final question from me is uh the are your treatment was dose dependent and uh do you think why the uh effect is dose dose dependent or if the treatment effect is dose dose dependent, do you want to increase the dose when you apply this technique to your clinical cases? Uh, I I I think they one minute they they can uh work scnic. So they uh they can improve the uh drug effect uh such as we combined the uh rapamycin and the steroid uh to treat the KH. And uh I think the the combined drugs maybe can work better than the single drugs. So any other comments or question from or from audience, if not, I think we can move on to the next session. Uh I guess we only have like 30 maybe 20, 30 seconds left but there was a question about doing a pre-clinical trial to assess the dosing and and and and get specific. Are you going to do 10 seconds a pre-clinical trial before an actual clinical trial? Uh, yes, of course, we we we have tried a few patients before we uh start a clinical trial. Yes. Okay. Great. I am so surprised I understood most of that because usually I don't understand anything basic science. So that was uh that was impressed I understood most of it.