Yong Wu, Ph.D. Division of Cancer Research and Training (DCRT) Charles R. Drew University of Medicine & Science
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1 Yong Wu, Ph.D. Division of Cancer Research and Training (DCRT) Charles R. Drew University of Medicine & Science Jay Vadgama, Ph.D Chief, Division of Cancer Research and Training
2 Background One in 8 women will develop breast cancer during the course of her life. Recent developments in the field of drug development have provided a significant increase in progression free survival and decrease in mortality of this disease. However, one of the main problems in breast cancer remains the development of resistance to current treatments and relapse of the disease.
3 What is triple negative breast cancers (TNBC)? A triple negative breast cancer (TNBC) diagnosis means that the offending tumor is estrogen receptor-negative (ER-), progesterone receptor-negative (PR-) and human epidermal growth factor receptor 2 (HER2)-negative (HER2-). These negative results mean that the growth of the cancer is not supported by the hormones estrogen and progesterone, nor by the presence of too many HER2 receptors. Therefore, triple-negative breast cancer does not respond to hormonal therapy (such as tamoxifen or aromatase inhibitors) or therapies that target HER2 receptors, such as Herceptin (chemical name: trastuzumab).
4 There is no known target for the treatment of TNBC subset, which is more aggressive than the other types of the disease. About 10-20% of breast cancers are found to be triple-negative. For doctors and researchers, there is intense interest in finding new medications that can treat this kind of breast cancer. Early studies are trying to find out whether certain medications can interfere with the processes that cause triple-negative breast cancer to grow.
5 Therefore, it is imperative to develop new drugs that can block the activity of pro-survival signaling within the cancer cells and inhibit their growth, particularly in the TNBC subset. In an attempt to identify such drugs, we screened a panel of Curcuminbased Compounds, 13 1,5-diheteroarylpenta-1,4-dien-3-ones (compounds 34 to 46), for their effectiveness in inhibiting the growth of both TNBC and ER+ breast cancers.
6 Curcumin Curcumin, extracted from Curcuma longa (turmeric) of the Zingiberaceae family, an ancient Indian herb used in curry powder, represents a typical example of dietary natural products displaying a wide range of interesting medicinal properties. The low cancer incidence in Asia has been, at least partly, demonstrated to be associated with the Asian diet (including turmeric). The anticancer potential of curcumin was first reported by Kuttan and coworkers in Curcumin has already entered several human clinical studies for potential prevention and treatment of cancer but its clinical development has been slown-down by its poor pharmacokinetic profile.
7 Structures of curcumin and its analogs
8 Results
9 Comp 34 Comp 45 Comp 39 Comp 44 Comp 46 Comp 42 Comp 36 Comp 43 Comp 37 Comp 41 Comp 40 Comp 35 Comp 38 IC50 nm MDA-MB MCF IC50 nm Comp 34 Comp 39 Comp 45 Comp 44 Comp 34 Comp 46 Comp 46 Comp 44 Comp 36 Comp 39 Comp 40 Comp 45 Comp 42 Comp 35 Comp 37 Comp 37 Comp 35 Comp 40 Compd 43 Comp 42 Comp 41 Comp 38 Comp 36 Comp 43 IC50 nm Comp 41 Comp T47D The IC 50 is a measure of how effective a drug is. It indicates how much of a particular drug is needed to inhibit a given biological process by half
10 Comp 34 Comp 45 Comp 39 Comp 44 Comp 46 Comp 42 Comp 36 Comp 43 Comp 37 Comp 41 Comp 40 Comp 35 Comp 38 IC50 nm MDA-MB-231 cell line treated with compounds
11 Con. Comp 34 Comp 35 Comp 36 Comp 37 Comp 38 Comp 39 Comp 40 Comp 41 Comp 42 Comp 43 Comp 44 Comp 45 Comp 46 MDA-MB-231 treated with compound DNA checkpoint associated kinases phosphorylate BRCA1 under conditions of DNA damage, making cells sensitive or resistant to various stresses. UV induced phosphorylation of particular residues of BRCA1 is crucial for activation of caspase 3. Mutant p53 in MDA-MB-231 breast cancer cells is stabilized by elevated phospholipase D activity and contributes to survival signals generated by phospholipase D p38 MAP Kinase Mediates Apoptosis through Phosphorylation of BimEL at Ser-65 p-brca1 p-p53 Total BRCA Total p53
12 Comp 34 Comp 39 Comp 45 Comp 44 Comp 46 Comp 36 Comp 40 Comp 42 Comp 37 Comp 35 Compd 43 Comp 41 Comp 38 IC50 nm IC50 = 8.3uM MCF-7 cell line treated with compounds
13 Con. Comp 34 Comp 35 Comp 36 Comp 37 Comp 38 Comp 39 Comp 40 Comp 41 Comp 42 Comp 43 Comp 44 Comp 45 Comp 46 MCF-7 treated with compound Wild type p53 in MCF-7 breast cancer cells induces cell cycle arrest and apoptosis p-brca1 p-p53 Total BRCA Total p53
14 Comp 34 Comp 46 Comp 44 Comp 39 Comp 45 Comp 35 Comp 37 Comp 40 Comp 42 Comp 41 Comp 38 Comp 36 Comp 43 IC50 nm IC50 = 6.1 μm T47D cell line treated with compounds
15 T47D treated with compound Con. Comp 34 Comp 35 Comp 36 Comp 37 Comp 38 Comp 39 Comp 40 Comp 41 Comp 42 Comp 43 Comp 44 Comp 45 Comp 46 T47D Cell line harbours a p53 missense mutation Mutant p53 mediates survival of breast cancer cells p-p53 P21 p-brca1 Total p53 Total BRCA ER+ T47D human breast cancer cells
16 Compound # 34 Control Map Growth assays and signaling effects of compounds MDA-MB-231 breast cancer cell lines
17 Changes in phosphoprotein levels induced by Comp #34 treatment % Change in phosphoprotein level p70 T389 TOR STAT3 HSP60 Akt T308 Src p53 S392 RSK1/2/3 ß-Catenin PRAS40 WNK1 p53 S406 p70 T421/S424 p53 S15 EGF AMPKa2 STAT2 Hck p38a Akt S473 STAT5a ERK1/2 MSK1/2 JNK GSK-3a/ß CREB Chk-2 Yes STAT5b STAT6 HSP27 c-jun FAK STAT5a/b
18 Genes with very low or not detectable level Changes in phosphoprotein levels induced by Comp #34 treatment Fyn AMPKa1 STAT3 p27 PLC-γ1 Lyn Lck Fgr PYK2 enos PDGF Log of % Change in phosphoprotein level
19 Con. 5 min. 15 min. 30 min. 1 hour 18 hours 6 hours 24 hours MDA-MB-231- Treatment with comp # 34 at different time points Comp # 34 Total - AKT p-mtor Total mtor p-p70 p-s6 p-p38 Total p-38 p-erk Total ERK p-p53 Total p-53 GAPDH
20 Con. 10 nm 50nM 250nM 1000nM MDA-MB-231- Treatment with different concentrations of Compound 34 p-p38 P38 Total mtor Total AKT P-ERK p-p53 Total P53 P-BRCA1 BRCA1 Tubulin
21 Con C # 34 C # 34 + MEKi MEKi Con C # 34 C # 34 + MEKi MEKi MDA-MB-231 cell line p-pi3k (p85/55) p-akt T308 p-gsk3α/β p-mtor Total AKT Total GSK Total mtor p-p70 p-s6 p-p38 Total p-38 p-c-jun p-p53 p-erk Total p53 Total ERK Tubulin Non specific band
22 mrna Down regulation of proteins by com#34 is not due to proteasome degradation Comp # 34 MG p-mtor p-p70 p-p38 p-p53 mtor P70 P38 P53
23 Acknowledgements Dr. Vadgama Dr. Yanyuan Wu Dr. Atefi Dr. Wang Dr. Wu Kevin kemp We acknowledge research support from UHI Research Award! Collaborators: Zhen Cheng, Ph.D. Associate Professor, Director of Cancer Molecular Imaging Chemistry Laboratory (CMICL) Stanford University Guangdi Wang, Ph.D., Professor of Chemistry Xavier s RCMI Cancer Research Center; Xavier University Qiao-Hong Chen, Ph.D. Assistant Professor (Tenured) California State University, Fresno, Fresno
24 Case-Based Question History: 33- year- old premenopausal woman with diagnosis of T1N1M0 breast cancer (left) Treated with breast conserving surgery and axillary lymphadenectomy No family history of breast cancer Pathology _ Invasive ductal carcinoma, grade 3 _ T size 1.2cm with clear margins _ Presence of lymphatic invasion _ ER and PR negative _ HER-2 negative with FISH _ Ki67: 60% Axillary lymph nodes 2/23 positive
25 QUESTION: Does the patient belong to a special category as far as risk of relapse is concerned? 1. No 2. Yes, but only due to her young age 3. Yes, but only due to the lymph nodes status 4. Yes, patients with triple negative tumors have higher risk of relapse but only the first three years after diagnosis 5. I do not know, I would evaluate the risk of relapse by Mammaprint or OncotypeDX
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