title: Novel targeting of PEGylated liposomes for codelivery of TGF-β1 siRNA and four antitubercular drugs to human macrophages for the treatment of mycobacterial infection: a quantitative proteomic study.
abstract: Tuberculosis (TB) is still a major public health issue in developing countries, and its chemotherapy is compromised by poor drug compliance and severe side effects. This study aimed to synthesize and characterize new multimodal PEGylated liposomes encapsulated with clinically commonly used anti-TB drugs with linkage to small interfering RNA (siRNA) against transforming growth factor-β1 (TGF-β1). The novel NP-siRNA liposomes could target THP-1-derived human macrophages that were the host cells of mycobacterium infection. The biological effects of the NP-siRNA liposomes were evaluated on cell cycle distribution, apoptosis, autophagy, and the gene silencing efficiency of TGF-β1 siRNA in human macrophages. We also explored the proteomic responses to the newly synthesized NP-siRNA liposomes using the stable isotope labeling with amino acids in cell culture approach. The results showed that the multifunctional PEGylated liposomes were successfully synthesized and chemically characterized with a mean size of 265.1 nm. The novel NP-siRNA liposomes functionalized with the anti-TB drugs and TGF-β1 siRNA were endocytosed efficiently by human macrophages as visualized by transmission electron microscopy and scanning electron microscopy. Furthermore, the liposomes showed a low cytotoxicity toward human macrophages. There was no significant effect on cell cycle distribution and apoptosis in THP-1-derived macrophages after drug exposure at concentrations ranging from 2.5 to 62.5 μg/mL. Notably, there was a 6.4-fold increase in the autophagy of human macrophages when treated with the NP-siRNA liposomes at 62.5 μg/mL. In addition, the TGF-β1 and nuclear factor-κB expression levels were downregulated by the NP-siRNA liposomes in THP-1-derived macrophages. The Ingenuity Pathway Analysis data showed that there were over 40 signaling pathways involved in the proteomic responses to NP-siRNA liposome exposure in human macrophages, with 160 proteins mapped. The top five canonical signaling pathways were eukaryotic initiation factor 2 signaling, actin cytoskeleton signaling, remodeling of epithelial adherens junctions, epithelial adherens junction signaling, and Rho GDP-dissociation inhibitor signaling pathways. Collectively, the novel synthetic targeting liposomes represent a promising delivery system for anti-TB drugs to human macrophages with good selectivity and minimal cytotoxicity.
authors: Department of Orthopedics, General Hospital of Tianjin Medical University, Tianjin, People's Republic of China ; Department of Spinal Surgery, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, People's Republic of China ; Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA.
authors: Ning-Kui
authors: NK
authors: Niu
authors: Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA.
authors: Juan-Juan
authors: JJ
authors: Yin
authors: Department of Colorectal Surgery, General Hospital of Ningxia Medical University, Yinchuan, Ningxia, People's Republic of China.
authors: Yin-Xue
authors: YX
authors: Yang
authors: Department of Orthopedics, General Hospital of Tianjin Medical University, Tianjin, People's Republic of China.
authors: Zi-Li
authors: ZL
authors: Wang
authors: Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA.
authors: Zhi-Wei
authors: ZW
authors: Zhou
authors: Guizhou Provincial Key Laboratory for Regenerative Medicine, Stem Cell and Tissue Engineering Research Center and Sino-US Joint Laboratory for Medical Sciences, Guizhou Medical University, Guiyang, Guizhou, People's Republic of China.
authors: Zhi-Xu
authors: ZX
authors: He
authors: Department of General Surgery, The First People's Hospital of Shunde Affiliated to Southern Medical University, Shunde, Foshan, Guangdong, People's Republic of China.
authors: Xiao-Wu
authors: XW
authors: Chen
authors: Research Center for Bioengineering and Sensing Technology, University of Science and Technology Beijing, Beijing, People's Republic of China.
authors: Xueji
authors: X
authors: Zhang
authors: School of Medicine, Deakin University, Waurn Ponds, VIC, Australia.
authors: Wei
authors: W
authors: Duan
authors: Department of Internal Medicine, University of Utah and Salt Lake Veterans Affairs Medical Center, Salt Lake City, UT, USA.
authors: Tianxin
authors: T
authors: Yang
authors: Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, Tampa, FL, USA.
authors: Shu-Feng
authors: SF
authors: Zhou
mesh_terms: D000995:Antitubercular Agents / Q000008:administration & dosage* / Q000494:pharmacology / Q000633:toxicity; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001343:Autophagy / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D016503:Drug Delivery Systems; D020868:Gene Silencing; D006801:Humans; D008081:Liposomes; D008264:Macrophages / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D040901:Proteomics / Q000379:methods; D034741:RNA, Small Interfering / Q000008:administration & dosage*; D015398:Signal Transduction / Q000187:drug effects / Q000235:genetics; D053773:Transforming Growth Factor beta1 / Q000235:genetics*; D014376:Tuberculosis / Q000188:drug therapy
chemical_list: D000995:Antitubercular Agents; D008081:Liposomes; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1; D011092:Polyethylene Glycols
keywords: NF-κB; SILAC; apoptosis; autophagy; cell cycle; cytokine; interleukin; liposome; proteomics; tuberculosis