课题名称Identification of synergy between FDA-approved cancer drugs and OGT inhibitors in breast cancer cells课题性质radic; 基础研究应用课题 设计型 调研综述 理论研究开题报告内容：（包括拟研究或解决的问题、采用的研究手段及文献综述，不少于2000字）Research purpose and significance:Breast cancer is the worlds most common cancer in women as second in overall mortality, triple negative breast cancer(TNBC) is highly aggressive, metabolic and basal-like subtype tumor accounting for 12-24% of all breast cancers,the features of TNBC are that estrogen receptor(ER) and progesterone receptor(PR) negative as a kind of breast cancer that overexpresses human epidermal growth factor receptor2(HEG2). Compared with non-TNBCs, a large proportion of TNBCs occurs in younger women, particularly in women of low socioeconomic status. As TNBC lacks effective target, treatment regimens often fail to slow tumor progression, there is a urgent need in developing effective and economical therapy in clinic.Now, combinatorial treatment is a kind of strategy to repurpose drugs that are already FDA approved instead of committing huge amounts of time and money to develop new drug, we can take two or more anti-tumor treatments and assess their ability to synergize with one another, and we can also improve the efficacy of a drug, or possibly reduce side effects by requiring a smaller dose. Through combinatorial treatment using two FDA approved drugs, we can effectively deal with the current TNBC clinical problems.Research topic at home and abroad:Hexosamine biosynthetic pathway(HBP) is a metabolic pathway that is altered in cancer cells, a product of this pathway, UDP-N-acetylgluosamine(UDP-GlcNAc), can be attached to various proteins post-translationally via the enzyme O-linked beta;-N-acetylglucosamine (O-GlcNAc) transferase (OGT).Total O-GlcNacylation and OGT levels have been shown to be elevated in breast cancer to promote oncogenesis and tumor progression, especially in TNBC. Some work has already shown that the targeting of this enzyme via RNAi or OGT inhibitors can be used to specifically kill breast cancer cells, while avoiding the killing of non-transformed mammary epithelial cells in vitro. But the effective dose of OGTi is big for clinical use, moreover, it will cause many side effect under that dose. Combinatorial treatment is another way to deal with TNBC. One research group have treated TNBC cell lines with the HDAC inhibitor romidepsin and the methyltransferase inhibitor decitabine, two epigenetic modifying drugs approved by the FDA for the treatment of various hematologic malignancies. Combinatorial treatment leads to synergistic inhibition of cell growth and induction of apoptosis above levels of individual drug treatments alone. Research aim:1.Test whether Romidepsin can selectively kill triple negative breast cancer cells.2.Test synergy between O-GlcNAcylation inhibitors and current FDA approved cancer drug Romidepsin in triple negative breast cancer cells.Research way:1.Get MDA-MB-453 MDA-MB-231 MDA-MB10X and MCF-7 cells and confirm that Romidepsin blocks cell growth via crystal violet and MTS assay. Do dose response. Night before plate 100,000 cells in 6 well plate. Next day add DMSO, 1 nM, 5 nM, 50 nM, 100 nM, 500 nM. Stain crystal violet in 48 hrs. Perform MTS assay with same doses/time.2.Repeat above and collect lysates. Perform western blot and probe effect of Romidepsin probe with p21, acetylated-H3. Examine apoptotic markers: Cleaved Caspses-3, PARP, BIM, BAX. 3.Infect and select MDA-MB-231 using OGT inhibitor, add DMSO, 1 nM, 5 nM, 10 nM Romidepsin to Control and OGT RNAi cells. Perform crystal violet.Research schedule：1. The first step (2/18-3/18)Learn how to do crystal violet tissue culture and western blotting, then culture MDA-MB-231, treat with Romidepsin, collect cell lysates, observe whether Romidepsin can block 231cells.2. The second step (3/18-5/18)Culture MDA-MB-453, MDA-MB10X and MCF-7 cells, do CV and WB, observe whether Romidepsin has selective anti-tumor function in breast cells.3. The third step (5/18-6/18)Treat MDA-MB-231with OGT inhibitor and Romidepsin, do CV and WB, observe whether they have anti-tumor synergy.Expected results:Results will be presented in the form of basic research papersReview:Targeting Triple-negative breast cancer using novel targets[Abstract]Triple-negative breast cancer(TNBC) is regarded as an aggressive disease that affects a young patient population and for which effective targeted therapy is not yet available. Intense efforts have been made to gain a better understanding of TNBC from the histologic to the genomic and molecular levels. This review will list several reported therapeutic potential of biomarkers and emerging targeted therapies. [Keyword] Triple-negative breast cancer(TNBC);O-GlcNAcylation; Histone deacetylase (HDAC) inhibitors Breast cancer is the worlds most common cancer in women as second in overall mortality,Approximately 12% to 24% of these cases are categorized as triple-negative breast cancer (TNBC)1. TNBC is defined immunohistochemically as breast cancer that does not over- express human epidermal growth factor receptor 2 (HER2, ERBB2) and is estrogen receptor (ER) and progesterone receptor (PR) negative. TNBC is regarded as an aggressive disease that affects a young patient population and for which effective targeted therapy is not yet available2-3. Thus common targets for non-TNBC is not available for TNBC, we list the three other available biomarkers and target therapies from literature. Further discovery and validation of biomarkers to serve as prognostic aids and as potential therapeutic targets are necessary to advance the treatment of patients with TNBC. O-linked beta;-N-acetylglucosamine (O-GlcNAc) transferase(OGT)One of the most well-known changes in the cellular energetics within cancer cells is the Warburg Effect3.The Warburg Effect is a phenomenon of cancer cells characterized by an increased utilization of glucose as a source of energy and biosynthetic intermediates. However, glycolysis is not the only metabolic pathway that is altered by the Warburg Effect; 3-5% of glucose that enters a cell is shunted through the hexosamine biosynthetic pathway (HBP)4.A product of this pathway, UDP-N-acetylglucosamine (UDP-GlcNAc), is the substrate for the enzyme O-linked beta;-N-acetylglucosamine (O-GlcNAc) transferase (OGT)5.This enzyme is involved in an important post-translational modification, O-GlcNacylation, that involves the transfer of UDP-GlcNAc to serine and threonine residues of select nuclear and cytosolic proteins.O-GlcNAcylations role is to respond to nutrient levels and stress by modulating cellular signaling and transcription regulatory pathways6.Total O-GlcNacylation and OGT levels have been shown to be elevated in breast cancer, and have been linked to oncogenesis and tumor progression. Some work has already shown that the targeting of this enzyme via RNAi or OGT inhibitors can be used to specifically kill breast cancer cells, while avoiding the killing of non-transformed mammary epithelial cells in vitro7. However, the reported effective dose of OGTi is 100 mu;M, too big for clinical use.moreover, it will cause many side effect under effective dose.Histone deacetylase (HDAC)Post‐translational modifications to the histones of chromatin play an important role in the regulation of gene expression. One reversible histone modification is non-histone and histone acetylation, which is regulated by histone acetyl transferases (HATs) and histone deacetylases (HDACs)8. There are currently 18 human HDACs that have been identified based on their function. Abnormal expression of these enzymes can contribute to carcinogenesis9. HDACs control numerous essential mechanisms during cell development and tissue maintenance in almost all cell types. Moreover, abnormal expression of HDACs has been found in a broad range of cancer types including breast cancer. Now researchers find that prevention of aberrant HDACs function by affecting HDAC expression, particularly overexpression, can be an attractive target for cancer therapy10. Currently, gt;20 HDAC inhibitors are undergoing preclinical and clinical tests, particularly vorinostat (Zolinza) and depsipeptide (Istodax), having been approved by the Food and Drug Administration (FDA) to be used for the treatment of T-cell lymphoma11-12. Although HDAC inhibitors are potent anticancer agents, certain studies have revealed that HDAC inhibitors can induce multidrug resistance (MDR), contributing to a poor prognosis in cancer treatment, this is an obstacle for the success of chemotherapy. Now, there is a few evidence shows that depsipeptide (Romidepsin) can also block the TNBC progression. This could be reuse of Romidepsin.Androgen Receptor Androgen receptor(AR) is coexpressed with ER in the majority of breast cancers12. AR inhibits proliferative activity in ER-positive tumors, but it acts independently to promote tumorigenesis in an androgen-dependent manner in ER-negative tumors like TNBC.AR can be present independent of ER, with one recent review estimating that AR is expressed in approximately 30% of TNBCs13. AR mediates the ligand-dependent activation of the WNT and HER2 signaling pathways, which is accomplished through direct transcriptional induction of WNT7B and HER3. Specific targeting of the AR with bicalutamide inhibited the growth of dihydrotestosterone-stimulated ER-negative/ HER2-positive tumors in vivo14. Bicalutamide efficacy was tested on TNBC cell lines, of which the LAR subtype cell lines were significantly more sensitive than the other sub- types15. Although the laboratory results are encouraging, the effects of AR antagonists in patients with breast cancer are currently unknown. In summary, three targets mentioned above have advantages and disadvantages for treating TNBC, however, if we can find the synergy between these and combine these into one strategy, we can also improve the efficacy of a drug, or possibly reduce side effects by requiring a smaller dose. I think combinatorial treatment is a kind of strategy to repurpose drugs instead of committing huge amounts of time and money to develop new drug, it may be the available direction for treating TNBC or other cancers.[Reference]1.Newman L A. Disparities in breast cancer and african ancestry: a global perspective[J]. Breast Journal, 2015, 21(2):133-9.2. Schmadeka R, Harmon B E, Singh M. Triple-negative breast carcinoma: current and emerging concepts[J]. American Journal of Clinical Pathology, 2014, 141(4):462-77.3.Alhajj M, Wicha M S, Benitohernandez A, et al. Prospective identification of tumorigenic breast cancer cells[J]. Proceedings of the National Academy of Sciences of the United States of America, 2003, 100(7):3983-3988.4. Xu X D, Shao S X, Jiang H P, et al. Warburg effect or reverse Warburg effect? A review of cancer metabolism.[J]. Oncol Res Treat, 2015, 38(3):117-22.5. Kim Y H, Nakayama T, Nayak J. Glycolysis and the Hexosamine Biosynthetic Pathway as Novel Targets for Upper and Lower Airway Inflammation:[J]. Allergy Asthma 69:6131-6140. 15.Doane AS, Danso M, Lal P, et al. An estrogen receptor- negative breast cancer subset characterized by a hormonally regulated transcriptional program and response to androgen. Oncogene. 2006;25:3994-4008. 16.McNamara KM, Yoda T, Takagi K, et al. Androgen receptor in triple negative breast cancer. J Steroid Biochem Mol Biol. 2013;133:66-76. 17.Ni M, Chen Y, Lim E, et al. Targeting androgen receptor in estrogen receptor-negative breast cancer. Cancer Cell.
Identification of synergy between FDA-approved cancer drugs and OGT inhibitors in breast cancer cells文献综述
2022-12-30 04:12
