The goals of our research are i) to better understand the genetic and molecular bases for variable drug response and drug interactions with a focus on membrane transporters; ii) to develop novel anticancer drugs targeting the proteasomes and their delivery strategies to achieve desirable pharmacokinetic and pharmacodynamic profiles; iii) to evaluate the pharmacokinetics and pharmacogenomics of anticancer drugs in early clinical trial settings and to assess the drug exposure and response in vivo using pharmacometric modeling and simulation.
I. Investigation of the impact of splicing and other genetic variations on drug transporters and proteasomes
Comparison of cancer-type and liver-type OATP1B3
The production of distinct mRNA transcripts from a single gene via alternative splicing is a common, yet important mechanism of generating proteomic diversity in eukaryotic cells. Alterations in splicing patterns have been increasingly associated with disease development and variable response to drug therapy. We have been investigating the functional significance and regulatory mechanisms of splicing variants of genes encoding membrane transporters or molecular targets for cancer therapy such as proteasomes.
Our group was the first to report that colon and pancreatic cancer cells express the cancer-type variant of OATP1B3 (ct-OATP1B3) with the distinct mRNA and protein identity from the liver-type OATP1B3 detected in non-malignant hepatocytes (Thakkar et al. Mol Pharm 2013). In our follow-up investigation, we identified hypoxia-inducible factor 1alpha (HIF-1alpha) as a positive regulator of ct-OATP1B3 expression (Han et al. Biochem Pharmacol 2013). Recently, we reported that the N-terminal region of OATP1B3 (which is present in liver-type OATP1B3, but absent in ct-OATP1B3) plays a crucial role in regulating membrane trafficking of lt-OATP1B3 (Chun et al. Biochem Pharamcol 2017).
We have also been investigating the impact of various genetic variations associated with different subunits that constitute the multi-subunit protease complex called proteasomes. The β1i subunit of the immunoproteasome harbors frequently occurring genetic variations (p.60R>H) and conflicting results had been reported with regard to its functional impact on the immunoproteasome activity. We reported that the codon 60 genetic variations of the β1i subunit do not account for variable expression/activity of the immunoproteasome (Park et al. PLoS One 2013). Currently we are investigating the impact of alternatively spliced transcripts of the proteasome subunits on the proteasome function.
- Thakkar N, Kim K, Jang ER, Han S, Kim K, Kim D, Merchant N, Lockhart AC, Lee W. A cancer-specific variant of the SLCO1B3 gene encodes a novel human organic anion transporting polypeptide 1B3 (OATP1B3) localized mainly in the cytoplasm of colon and pancreatic cancer cells. Mol Pharm. 2013;10(1):406-416. (PMID:23215050)
- Han S*, Kim K*, Thakkar N*, Kim D, Lee W. Role of Hypoxia Inducible Factor-1α in the Regulation of the Cancer-Specific Variant of Organic Anion Transporting Polypeptide 1B3 (OATP1B3) in Colon and Pancreatic Cancer. Biochem Pharmacol 2013;86:816-823 (*, equally contributed) (PMID: 23924606)
- Thakkar N, Lockhart AC, Lee W. Role of Organic Anion Transporting Polypeptides (OATPs) in Cancer Therapy, AAPS J. 17(3):535-45 (2015) (PMID: 25735612)
- Chun S-E, Thakkar N, Oh Y, Park JE, Han S, Ryoo G, Hahn H, Maeng SH, Lim Y-R, Han BW, Lee W The N-terminal region of Organic Anion Transporting Polypeptide 1B3 (OATP1B3) plays an essential role in regulating its plasma membrane trafficking. Biochem Pharmacol 131:98-105 (2017/05) (PMID: 28216016)
- Park JE, Ryoo G, Lee W. Alternative Splicing: Expanding Diversity in Major ABC and SLC Drug Transporters. AAPS J (accepted, 2017)
- Park JE, Ao L, Miller Z, Kim K, Wu Y, Jang ER, Lee EY, Kim KB, Lee W. PSMB9 codon 60 polymorphisms have no impact on the activity of the immunoproteasome catalytic subunit β1i expressed in multiple types of solid cancer. PLoS One 8(9):e73732 (2013) (PMID: 24040045)
II. Development of novel proteasome inhibitor drugs and delivery strategies to improve anticancer efficacy and expand therapeutic utilities
FDA-approved proteasome inhibitor drugs
The proteasome is an effective anticancer target, firmly validated by the three FDA-approved drugs, bortezomib (Velcade®), carfilzomib (Kyprolis®) and ixazomib (Ninlalo®). These drugs have transformed the treatment landscape for multiple myeloma and other hematological malignancies. In particular, the second-generation proteasome inhibitor drug carfilzomib has placed itself as part of the frontline multiple myeloma therapy, thanks to its much improved safety and efficacy profiles over bortezomib. Despite these remarkable successes, carfilzomib has shown limited efficacy in patients with solid cancers, possibly due to its short half-life in blood and insufficient access of active drug to tumor tissues. Working together with experts in the field of drug delivery (Drs. Yoonsoo Bae, University of Kentucky; Dr. Yoon Yeo, Purdue University), we are developing novel nano-formulations for carfilzomib that can improve the biopharmaceutical properties and the anticancer efficacy in multiple myeloma and other types of cancers (Ao et al. J Pharm Exp Ther 2015; Park et al. PLoS One, 2017).
We also collaborate with scientists who have the expertise and experience in the development of novel proteasome inhibitors (Dr. Kyung Bo Kim, University of Kentucky). Our goal is to develop novel proteasome inhibitor drugs that can overcome the biopharmaceutical limitations of existing proteasome inhibitor drugs. Like many cancer therapeutics, the proteasome inhibitor drugs are subject to de novo or acquired resistance, which is a major clinical obstacle. We have been investigating the molecular mechanisms underlying cancer resistance to carfilzomib and potential strategies to overcome resistance (Ao et al. Mol. Pharmaceutics 2012). A better mechanistic understanding of cancer resistance to proteasome inhibitors has been utilized to screen and develop novel next-generation proteasome inhibitor drugs that can be effective in patients who do not have any further therapeutic options (Miller et al. J Med Chem 2015). We also investigate the biological impact and therapeutic potential of novel inhibitors for the immunoproteasome (an alternative type of proteasome that is often unregulated in cancer cells).
- Ao L, Reichel D, Hu D, Jeong H-Y, Kim KB, Bae Y, Lee W. Polymer micelle formulations of proteasome inhibitor carfilzomib for improved metabolic stability and anti-cancer efficacy in human multiple myeloma and lung cancer cell lines. J Pharm Exp Ther 355(2):168-73. (doi: 10.1124/jpet.115.226993; PMID: 26311812).
- Reichel D, Lee MJ, Lee W, Kim KB, Bae Y Tethered polymer nanoassemblies for sustained carfilzomib release and prolonged suppression of proteasome activity, Ther Deliv 7(10):665-681 (2016) (PMID 27790952).
- Park J, Chun S-E, Reichel D, Min JS, Lee S-C, Han S, Ryoo G, Oh Y, Park S-H, Ryu H-M, Kim KB, Lee H-Y, Bae SK, Bae YB, Lee W Polymer micelle formulation for the proteasome inhibitor drug carfilzomib: Anticancer efficacy and pharmacokinetic studies in mice. PLoS One 12(3):e0173247 (2017.03) (PMID: 28273121)
- Miller Z, Kim KS, Lee DM, Kasam V, Baek SE, Lee KH, Zhang YY, Ao L, Carmony K, Lee NR, Zhou S, Zhao Q, Jang Y, Jeong HY, Zhan CG, Lee W, Kim DE, Kim KB Proteasome inhibitors with pyrazole scaffolds from structure-based virtual screening. J Med Chem 58(4):2036-41 (2015) (PMID: 25658656)
- Miller Z, Lee W, Kim KB. The Immunoproteasome as a Therapeutic Target for Hematological Malignancies, Current Cancer Drug Targets, 14(6):537-48 (2014). (PMID: 25059201)
III. Clinical Pharmacokinetics, Pharmacogenomics & Pharmacometrics
Interindividual differences in drug response and toxicities are consistently observed with most chemotherapeutic agents or regimens and many clinical variables (e.g., age, gender, diet, drug-drug interactions) affect drug responses. In particular, inherited variations in drug disposition (metabolism and transport) and drug target genes are known to substantially contribute to the observed variability in cancer treatment outcome. In our collaborative Phase II clinical study, we investigated the clinical utility of pharmacogenomically selected treatment using genetic polymorphisms in patients with gastric and gastroesophageal junction (GEJ) cancer (Goff et al. PLoS One 2014). Working together with clinical investigators, we investigate the pharmacokinetics of novel cancer drugs, drug formulations or new combination regimens in early clinical trial settings. Our group is also interested in pharmacometric modeling and simulation using preclinical and clinical data.
- Goff LW, Thakkar N, Du L, Chan E, Tan BR, Cardin DB, McLeod HL, Berlin JD, Zehnbauer B, Fournier C, Picus J, Wang-Gillam A, Lee W, Lockhart AC. Thymidylate Synthase Genotype-Directed Chemotherapy for Patients with Gastric and Gastroesophageal Junction Cancers, PLOS ONE, 18;9(9):e107424 (2014). (PMID: 25232828)
- Wang-Gillam A, Thakkar N, Lockhart AC, Williams K, Baggstrom M, Naughton M, Suresh R, Ma C, Tan B, Lee W, Jiang X, Mwandoro T, Trull L, Belanger S, Creekmore AN, Gao F, Fracasso PM, Picus J. A phase I study of pegylated liposomal doxorubicin and temsirolimus in patients with refractory solid malignancies. Cancer Chemother Pharmacol 74(2):419-26 (2014) (PMID: 24916546).
- Michel L, Ley J, Wildes TM, Schaffer A, Robinson A, Chun S-E, Lee W, Lewis Jr. J, Trinkaus K, Adkins D Phase I Trial of Palbociclib, a Selective Cyclin Dependent Kinase 4/6 Inhibitor, in Combination with Cetuximab in Patients with Recurrent/Metastatic Head and Neck Squamous Cell Carcinoma, Oral Oncol 58:41-8. doi: 10.1016/j.oraloncology.2016.05.011 (2016) (PMID: 27311401)
- Asaumi R, Toshimoto K, Tobe Y, Hashizume K, Nunoya K, Imawaka H, Lee W, Sugiyama Y. Comprehensive PBPK model of rifampicin for quantitative prediction of complex drug-drug interactions: CYP3A/2C9 induction and OATP inhibition effects. CPT: Pharmacometrics Syst Pharmacol (2018, in press) (PMID 29368402)
- Sato M, Toshimoto K, Tomaru A, Yoshikado T, Hisaka A, Lee W, Sugiyama Y. Physiologically based pharmacokinetic modeling of bosentan identifies the saturable hepatic uptake as A major contributor to its nonlinear pharmacokinetics. Drug Metab Dispos (2018, in press) (PMID: 29475833)