 My name is Balraj Singh, I am at MD Anderson Cancer Center in Houston, Texas. I'm going to talk about our paper published in OncoTarget, entitled, inhibition of resistant triple-negative breast cancer cells with low dose, 6-mal-cap-typurein, and 5-age acidity. As a background, our major goal is to improve therapy options in resistant cancers, such as TNBC and inflammatory-based cancer. It will involve overcoming deep, intrinsic resistance, which is caused by rare but adaptable cancer cells that can survive all the selection pressures in the body, often in quiescence. Our specific objective is to develop a more predictive cell culture model of resistance for evaluating therapy. A big limitation of cancer cell lines in cell culture is that artificially-rich culture medium promotes proliferation of most cells that are not fit to survive in the body. To address this limitation, we subject the aggressive TN-IBC cell lines to severe metabolic challenge by switching them to a glutamine-free medium for several weeks. This selection works well, eradicating more than 99.9 percent cells within few days. Rare cells survive in quiescence for several weeks, and then at 3 to 5 weeks they begin to proliferate, yielding colonies, and such cells can then grow indefinitely. Interestingly, cell lines developed from more aggressive disease such as TN-IBC are far much better under this challenge than other TNBC cell lines, such as MDA 231. Surprisingly, the cells selected in this manner are not only metabolically adaptable, but they have a variety of adaptability traits. Briefly, they are resistant to chemotherapy drugs, highly tumor-genic in nude mice, and importantly, they metastasize to multiple organs in nude mice, such as lungs, skin, brain, and at the molecular level, they are more stem-like based on the gene expression which indicates a high epithelial to mesenchymal transition. They carry a variety of genomic and transcriptomic alterations and post-transcriptional RNA modifications. Our priority is to investigate non-cytotoxic drugs that would overcome therapy resistance. For realistic air evaluation, we test low doses of drugs for long periods to determine whether the resistant cells are being inhibited or modified from resistant to sensitive. How did we choose the drugs that we evaluated in this study? We have found that RNA modifications such as alternate RNA splicing and a variety of base modifications affecting messenger RNA, ribosomal RNA, and TRNA, many of them are... We are seeing many of these modifications in these resistant cells. So we think that they may be playing an important role in the resistance phenotype, particularly their persistence and quiescence. So since there are many RNA-based modifications, instead of targeting individual modifications, we chose broadly active ribonucleoside analogs that may counter the effects of many of these modifications simultaneously. Looking for relatively safe drugs at low dose, we chose 6MP and 5-hazel cytodine, which are purine and premedine analogs respectively. 6MP is a common drug used in many autoimmune diseases such as ulcerative colitis, and it has excellent safety profile, and also it can keep the disease in remission for long times. 5-hazel cytodine is used as a maintenance drug in leukemia, again indicating that the low dose may be safe. At safety, these clinical studies indicate that these drugs may affect stem-like cells since they cause remission. In all these diseases as well, the stem-like cells are the root cause of the disease, and they need to be controlled for getting the remission. We found that a long treatment with low-dose 6MP inhibits resistant breast cancer cells in our model. It's analogous to the situation in ulcerative colitis, where 6MP takes several weeks for it to cause remission. Further, a prior treatment with 5-hazel cytodine sensitizes cells to inhibit with 6MP. So these results indicate that if one drug is not enough, both drugs could be combined. Finally, when the cells that are surviving after treatment with one drug or both drugs, we wanted to know whether they are sensitive to the chemotherapy drugs or not. What we found is that as compared to the untreated cells, they are more sensitive. Overall, our results indicate that low-dose 6MP may be a good candidate for clinical evaluation to halt relapse in high-risk TNBC and IBC. It may also be an attractive drug for combining with immune checkpoint blockade. Immune checkpoint blockade, which is increasingly being used in TNBC, the problem is it causes a lot of autoimmunity type of reactions. Results are not obtained. The benefit that the immune checkpoint blockade could be achieved is hindered by the autoimmunity. So 6MP may proactively inhibit the development of autoimmunity besides inhibiting a stem-like cancer cell, thus improving the outcomes from immune checkpoint blockade. Finally, besides the specific evaluation, this paper provides proof of concept that this is a good approach for investigating deep, intrinsic resistance and ways to overcome it. Next, we will tax the concept in other cancers besides breast cancer. We will evaluate compounds like 6MP in combination with immune checkpoint blockade. As I said, since the immune checkpoint blockade is being very widely used in many different cancers and we are hoping that these studies will lead to clinical trials. Finally, a word about OncoTarget. We have published three papers in OncoTarget. We have had very positive experience. The journal provided fair, good quality, fast review for the papers and importantly, it allows us to present the work the way we see it. Thank you.