Engineering polycation and lipid nanoparticle-based non-viral nanoparticles for delivery of nucleic acid therapeutics via systemic, local, or oral administration

We have developed a complex coacervation method for preparing DNA nanoparticles using natural polymers including chitosan (Roy, et al, 1999; Mao, et al., 2001) and gelatin (Truong-Le, et al, 1999), tailor-designed biodegradable polyphosphoesters (Wang, et al, 2001), block or graft copolymers with improved control over their self-assembly with plasmid DNA (Jiang, et al, 2010; 2011; 2012; 2013; Qu, et al, 2015) or siRNA (Shyam, et al, 2015) forming micellar nanoparticles. We have also developed approaches to tune the shapes of DNA nanoparticle with diameters in 20 – 100 nm range (Jiang, et al, 2013), and provided insights in the mechanism in shape regulation during the process. We also applied these gene therapy strategies for mucosal gene delivery and genetic immunization (Chew, et al, 2003; He, et al, 2018; Nie, et al, 2019), to enhance liver-targeted gene delivery of DNA nanoparticles via retrograde intrabiliary infusion (RII) method (Dai, et al, 2006; 2011) and probed the DNA transport kinetics of DNA nanoparticles using a SPECT/CT imaging method (Patil, et al, 2011). Recently, we have also developed a lipid nanoparticle screening platform to identify effective and cell-type specific compositions for the delivery of plasmid DNA and other nucleic acid therapeutics (Zhu, et al., 2022), and applied these lipid nanoparticles for liver-specific gene delivery and oral vaccination.

Representative Publications:

  1. Mao HQ, Roy K, Troung-Le VL, Janes KA, Lin KY, Wang Y, August JT, Leong KW. Chitosan-DNA nanoparticles as gene carriers: synthesis, characterization and transfection efficiency. Journal of Controlled Release. 70(3): 399–421 (2001). PMID: 11182210.
  2. Roy K, Mao HQ, Huang SK, Leong KW. Oral gene delivery with chitosan-DNA nanoparticles generates immunologic protection in a murine model of peanut allergy. Nature Medicine. 5(4): 387–391 (1999). PMID: 10202926.
  3. Patil RR, Yu J, Banerjee SR, Ren Y, Leong D, Jiang X, Pomper M, Tsui B, Kraitchman DL, Mao HQ. Probing in vivo trafficking of polymer/DNA micellar nanoparticles using SPECT/CT imaging. Molecular Therapy. 19(9): 1626–1635 (2011). PMCID: PMC3182352; PMID: 21750533.
  4. Williford JM, Archang MM, Minn I, Ren Y, Wo M, Vandermark J, Fisher PB, Pomper MG, Mao HQ. Critical length of PEG grafts on lPEI/DNA nanoparticles for efficient in vivo delivery. ACS Biomaterials Science & Engineering. 2(4): 567–578 (2016). PMID: 27088129.
  5. Zhu Y, Shen R, Vuong I, Reynolds RA, Shears MJ, Yao Z, Hu Y, Cho WJ, Kong J, Reddy SK, Murthy SC, Mao HQ. Multi-step screening of plasmid DNA/lipid nanoparticles and co-delivery with siRNA to enhance and prolong gene expression in the liver. Nature Communications. 13(1): 4282. doi: 10.1038/s41467-022-31993-y. PMID: 35879315; PMCID: PMC9310361 (2022).

Kinetically controlled assembly and scalable production of nanoparticles for delivery of biologic therapeutics

We have invented a new flash nanocomplexation (FNC) process for scalable production of polyelectrolyte complex nanoparticles (US patent issued in 2019); and applied this method to kinetically control the packaging of various type of nucleic acids (Santos, et al, 2016), protein therapeutics (He, et al, 2017, 2018, Nie, et al, 2019, Hanwright et al, 2022), small molecular weight drugs (Li, et al, 2022), and vaccines (Qiao, et al, 2018). Recently, we have extended this method to assemble uniform, stable, and off-the-shelf particles within the size range of 200 to 900 nm to enhance the co-delivery efficiency of different virus packaging plasmids, and improve the lentivirus production efficiency (Hu, et al, 2021).

Representative Publications:

  1. Santos JL, Ren Y, Vandermark J, Archang MM, Williford JM, Liu H, Lee J, Wang TH, Mao HQ. Continuous production of discrete plasmid DNA-polycation nanoparticles using flash nanocomplexation. Small. 12(45): 6214–6222 (2016). PMCID: PMC5149445; PMID: 27717227.
  2. He Z, Santos JL, Tian H, Huang H, Hu Y, Liu L, Leong KW, Chen Y, Mao HQ. Scalable fabrication of size-controlled chitosan nanoparticles for oral delivery of insulin. Biomaterials. 130: 28–41 (2017). PMID: 28359018.
  3. Hu Y, He Z, Hao Y, Gong L, Pang M, Howard GP, Ahn HH, Brummet M, Chen K, Liu HW, Ke X, Zhu J, Anderson CF, Cui H, Ullman CG, Carrington CA, Pomper MG, Seo JH, Mittal R, Minn I, Mao HQ. Kinetic control in assembly of plasmid DNA/polycation complex nanoparticles. ACS Nano. 13 (9): 10161–10178 (2019). PMCID: PMC7293580; PMID: 31503450.
  4. Hu Y, Zhu Y, Sutherland ND, Wilson DR, Pang M, Liu E, Staub JR, Berlinicke CA, Zack DJ, Green JJ, Reddy SK, Mao HQ. Size-controlled and shelf-stable DNA particles for production of lentiviral vectors. Nano Lett. 21(13): 5697–5705 (2021). PMCID: PMC8283758; PMID: 34228937.
  5. Hanwright PJ, Qiu C, Rath J, Zhou Y, von Guionneau N, Sarhane KA, Harris TGW, Howard GP, Malapati H, Lan MJ, Reddy S, Hoke A, Mao HQ, Tuffaha SH. Sustained IGF-1 delivery ameliorates effects of chronic denervation and improves functional recovery after peripheral nerve injury and repair. Biomaterials. 280: 121244 (2022). PMID: 34794826.
  6. Li S, Hu Y, Li A, Lin J, Hsieh K, Schneiderman Z, Zhang P, Zhu Y, Qiu C, Kokkoli E, Wang TH, Mao HQ. Payload distribution and capacity of mRNA lipid nanoparticles. Nature Communications. 13(1): 5561. doi: 10.1038/s41467-022-33157-4. PMID: 36151112; PMCID: PMC9508184 (2022).