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Enhancing Tumor Cell Response to Chemotherapy Through Nanoparticle-Mediated Codelivery of Sirna and Cisplatin Prodrug

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Enhancing Tumor Cell Response to Chemotherapy Through Nanoparticle-Mediated Codelivery of Sirna and Cisplatin Prodrug Enhancing tumor cell response to chemotherapy through nanoparticle-mediated codelivery of siRNA and cisplatin prodrug Xiaoyang Xua,b,1, Kun Xiec,1, Xue-Qing Zhanga,1, Eric M. Pridgend, Ga Young Parke, Danica S. Cuia, Jinjun Shia, Jun Wua, Philip W. Kantofff, Stephen J. Lipparde, Robert Langerb,d, Graham C. Walkerc,2, and Omid C. Farokhzada,2 aLaboratory of Nanomedicine and Biomaterials, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115; bThe David H. Koch Institute for Integrative Cancer Research and Departments of cBiology, dChemical Engineering, and eChemistry, Massachusetts Institute of Technology, Cambridge, MA 02139; and fDepartment of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA 02215 Edited* by Chad A. Mirkin, Northwestern University, Evanston, IL, and approved September 14, 2013 (received for review March 1, 2013) Cisplatin and other DNA-damaging chemotherapeutics are widely forming chemotherapeutics cause DNA damage as their primary used to treat a broad spectrum of malignancies. However, their mechanism of cellular cytotoxicity. However, several cellular application is limited by both intrinsic and acquired chemoresist- pathways are activated in response to their interaction with ance. Most mutations that result from DNA damage are the DNA, which include DNA repair pathways that remove the consequence of error-prone translesion DNA synthesis, which damage and translesion DNA synthesis (TLS) by specialized could be responsible for the acquired resistance against DNA- DNA polymerases that helps the cells tolerate the DNA damage damaging agents. Recent studies have shown that the suppression (4, 5). The Rev1/Rev3L/Rev7-dependent error-prone TLS path- of crucial gene products (e.g., REV1, REV3L) involved in the error- way has been shown to play an important role in cisplatin-induced prone translesion DNA synthesis pathway can sensitize intrinsi- mutations that improve the capacity of tumor cells to either repair cally resistant tumors to chemotherapy and reduce the frequency or tolerate DNA damage, resulting in acquired chemoresistance of acquired drug resistance of relapsed tumors. In this context, (6). Rev1 is a translesion DNA polymerase, while Rev3 is the combining conventional DNA-damaging chemotherapy with siRNA- catalytic subunit of the translesion DNA polymerase Polζ (Rev3L/ based therapeutics represents a promising strategy for treating Rev7). Recent studies using mouse lymphoma and lung cancer MEDICAL SCIENCES patients with malignancies. To this end, we developed a versatile models have shown that the suppression of error-prone TLS ac- nanoparticle (NP) platform to deliver a cisplatin prodrug and REV1/ tivity in mammalian cells by knocking down Rev1 or Rev3L can REV3L-specific siRNAs simultaneously to the same tumor cells. NPs inhibit drug-induced mutagenesis so that relapsed tumors remain are formulated through self-assembly of a biodegradable poly(lac- sensitive to subsequent treatment (6, 7). It has been suggested tide-coglycolide)-b-poly(ethylene glycol) diblock copolymer and that combining conventional chemotherapy with newly emerg- a self-synthesized cationic lipid. We demonstrated the potency ing siRNA therapeutics could be a promising strategy for fi of the siRNA-containing NPs to knock down target genes ef - ENGINEERING ciently both in vitro and in vivo. The therapeutic efficacy of NPs Significance containing both cisplatin prodrug and REV1/REV3L-specific siRNAs was further investigated in vitro and in vivo. Quantitative real- The development of acquired chemoresistance is often a clini- fi time PCR results showed that the NPs exhibited a signi cant and cal problem limiting the successful treatment of cancers. RNAi sustained suppression of both genes in tumors for up to 3 d after is showing promising results in human clinical trials. The com- a single dose. Administering these NPs revealed a synergistic ef- bination of chemotherapy with RNAi approaches to suppress fect on tumor inhibition in a human Lymph Node Carcinoma of the the expression of proteins involved in the emergence of drug Prostate xenograft mouse model that was strikingly more effec- resistance represents a promising synergistic strategy to cir- tive than platinum monotherapy. cumvent or reverse acquired chemoresistance. Such combina- tion therapy approaches require specific delivery vehicles to siRNA delivery | chemosensitivity | combination therapy encapsulate and deliver chemotherapy and siRNA therapeutics simultaneously in a controlled manner. Herein, we describe dvances in genomics and cell biology have highlighted the a nanoparticle technology to codeliver a DNA-damaging che- Aheterogeneity and complexity of cancer. It is generally ac- motherapeutic and siRNAs that impair the cell’s ability to repair cepted that cancer is usually the result of a combination of the DNA damage. This combination therapeutic approach can interconnected disease pathways that may not be treated effec- sensitize cancer cells to chemotherapeutics and shows superior tively with 1D therapeutic mechanisms (1). The inhibition of tumor inhibition compared with monochemotherapy. a pathway by a single-drug therapy often results in the emer- gence of drug resistance and tumor relapse, largely because of Author contributions: X.X., K.X., X.-Q.Z., R.L., G.C.W., and O.C.F. designed research; X.X., K.X., X.-Q.Z., E.M.P., D.S.C., J.S., and J.W. performed research; X.X., X.-Q.Z., G.Y.P., and S.J.L. pathway redundancy, cross-talk, compensatory and neutralizing contributed new reagents/analytic tools; X.X., K.X., X.-Q.Z., R.L., G.C.W., and O.C.F. ana- actions, and antitarget activities that commonly occur with sin- lyzed data; and X.X., K.X., X.-Q.Z., P.W.K., R.L., G.C.W., and O.C.F. wrote the paper. gle-drug cancer therapy (2). In some cases, relapse can result in Conflict of interest statement: O.C.F. and R.L. disclose their financial interest in BIND the emergence of phenotypically distinct and possibly more vir- Biosciences, Selecta Biosciences, and Blend Therapeutics, three biotechnology companies ulent tumors. For example, treatment of prostatic adenocarci- developing nanoparticle technologies for medical applications. P.W.K. and S.J.L. disclose their financial interest in BIND Biosciences and Blend Therapeutics. BIND, Selecta, and noma with androgen ablation therapies, such as abiraterone or Blend did not support the aforementioned research, and these companies currently have enzalutamide, results in the development of abiraterone or enza- no rights to any technology or intellectual property developed as part of this research. lutamide refractory castration-resistant prostate cancer that is *This Direct Submission article had a prearranged editor. phenotypically nonadenocarcinoma and represents a rare and 1X.X., K.X., and X.-Q.Z. contributed equally to this work. often lethal form of prostate cancer with a neuroendocrine phe- 2To whom correspondence may be addressed. E-mail: [email protected] notype (3). or [email protected]. Platinum agents are among the most widely used cytotoxic This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. agents for cancer therapy. Cisplatin and other DNA adduct- 1073/pnas.1303958110/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1303958110 PNAS Early Edition | 1of6 Downloaded by guest on September 25, 2021 improving the efficacy of chemotherapy through additive or characteristics and metabolic targets, to prostate cancer cells. synergistic effects (8). However, there remains a pressing need to engineer nanocarriers Since the discovery of RNAi, synthetic siRNA has emerged as that are capable of delivering combination therapeutics involving a class of attractive therapeutics for treatment of various dis- siRNA because systemic delivery of siRNA still remains chal- eases, including cancer (9, 10). Given the ability to target and lenging. Herein, we describe an integrated nanodelivery sys- silence nearly any gene of interest, specific siRNA can be con- tem capable of simultaneously delivering cisplatin prodrug and structed to target genes encoding proteins involved in DNA re- siRNAs against REV1 and REV3L to enhance chemosensitivity pair and the acquisition of multidrug resistance (6, 11). Naked of tumors. PLGA-PEG was formulated with a cationic lipid-like siRNA cannot readily cross cellular membranes due to its poly- molecule designated as G0-C14 into NPs that comprise three anionic and macromolecular characteristics, and it is susceptible components: an aqueous inner core, a cationic and hydrophobic to degradation by endogenous enzymes (12). Therefore, con- layer composed of PLGA and G0-C14, and a hydrophilic PEG siderable efforts have been made to develop safe and effective corona (Fig. 1A). The G0-C14 compound is synthesized with cat- vehicles to facilitate the delivery of siRNA into cells (13–15). ionic head groups that can efficiently bind siRNA via electrostatic Similarly, the methods by which chemotherapeutics are delivered interactions and flexible hydrophobic tails for self-assembly with also have a significant effect on the efficacy (16, 17). Recent PLGA-PEG to form Pt(IV)-prodrug encapsulating NPs (Fig. 1A). research has begun to explore the feasibility of combining che- In this study, we applied a Pt(IV)-prodrug approach previously motherapeutics with siRNA using a variety of nanocarrier plat- used in our laboratory to deliver cisplatin (26). In this approach, forms (18, 19). One of the earliest efforts using this therapeutic
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