Nucleic acids have gained recognition as promising immunomodulatory therapeutics. However, their potential is limited by several drug delivery barriers, and there is a need for technologies that enhance intracellular delivery of nucleic acid drugs. Furthermore, controlled and sustained release is a significant concern, as the kinetics and localization of immunomodulators can influence resultant immune responses. Here, we describe the design and initial evaluation of poly(lactic-co-glycolic) acid (PLGA) microparticle (MP) depots for enhanced retention and sustained release of endosomolytic nanoparticles that enable the cytosolic delivery of nucleic acids.
Endosomolytic p[DMAEMA]10kD-bl-[PAA0.3-co-DMAEMA0.3-co-BMA0.4]25kD diblock copolymers were synthesized by reversible addition-fragmentation chain transfer polymerization. Polymers were electrostatically complexed with nucleic acids and resultant nanoparticles (NPs) were encapsulated in PLGA MPs. To modulate release kinetics, ammonium bicarbonate was added as a porogen. Release profiles were quantified in vitro and in vivovia quantification of fluorescently-labeled nucleic acid. Bioactivity of released NPs was assessed using small interfering RNA (siRNA) targeting luciferase as a representative nucleic acid cargo. MPs were incubated with luciferase-expressing 4T1 (4T1-LUC) breast cancer cells in vitro or administered intratumorally to 4T1-LUC breast tumors, and silencing via RNA interference was quantified via longitudinal luminescence imaging.
Endosomolytic NPs complexed to siRNA were effectively loaded into PLGA MPs and release kinetics could be modulated in vitro and in vivovia control of MP porosity, with porous MPs exhibiting faster cargo release. In vitro, release of NPs from porous MP depots enabled sustained luciferase knockdown in 4T1 breast cancer cells over a five-day treatment period. Administered intratumorally, MPs prolonged the retention of nucleic acid within the injected tumor, resulting in enhanced and sustained silencing of luciferase relative to a single bolus administration of NPs at an equivalent dose.
This work highlights the potential of PLGA MP depots as a platform for local release of endosomolytic polymer NPs that enhance the cytosolic delivery of nucleic acid therapeutics.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Scanning electron microscopy
Ahn, J., T. Xia, A. Rabasa Capote, D. Betancourt, and G. N. Barber. Extrinsic phagocyte-dependent STING signaling dictates the immunogenicity of dying cells. Cancer Cell 33(5):862–873e5, 2018.
Ali, O. A., N. Huebsch, L. Cao, G. Dranoff, and D. J. Mooney. Infection-mimicking materials to program dendritic cells in situ. Nat. Mater. 8(2):151–158, 2009.
Ali, O. A., C. Verbeke, C. Johnson, R. W. Sands, S. A. Lewin, D. White, E. Doherty, G. Dranoff, and D. J. Mooney. Identification of immune factors regulating antitumor immunity using polymeric vaccines with multiple adjuvants. Cancer Res. 74(6):1670–1681, 2014.
Aliabadi, H. M. Natural polymers in nucleic acid delivery. In: Polymers and Nanomaterials for Gene Therapy, edited by R. Narain. Cambridge: Woodhead Publishing, 2016, pp. 55–80.
Aliru, M. L., J. E. Schoenhals, B. P. Venkatesulu, C. C. Anderson, H. B. Barsoumian, A. I. Younes, K. M. Ls, M. Soeung, K. E. Aziz, J. W. Welsh, and S. Krishnan. Radiation therapy and immunotherapy: what is the optimal timing or sequencing? Immunotherapy 10(4):299–316, 2018.
Amar-Lewis, E., A. Azagury, R. Chintakunta, R. Goldbart, T. Traitel, J. Prestwood, D. Landesman-Milo, D. Peer, and J. Kost. Quaternized starch-based carrier for siRNA delivery: from cellular uptake to gene silencing. J. Control. Release 185:109–120, 2014.
Arany, S., D. S. Benoit, S. Dewhurst, and C. E. Ovitt. Nanoparticle-mediated gene silencing confers radioprotection to salivary glands in vivo. Mol. Ther. 21(6):1182–1194, 2013.
Aznar, M. A., N. Tinari, A. J. Rullan, A. R. Sanchez-Paulete, M. E. Rodriguez-Ruiz, and I. Melero. Intratumoral delivery of immunotherapy-act locally, think globally. J. Immunol. 198(1):31–39, 2017.
Bartlett, D. W., and M. E. Davis. Insights into the kinetics of siRNA-mediated gene silencing from live-cell and live-animal bioluminescent imaging. Nucleic Acids Res. 34(1):322–333, 2006.
Beyranvand Nejad, E., M. J. Welters, R. Arens, and S. H. van der Burg. The importance of correctly timing cancer immunotherapy. Expert Opin. Biol. Ther. 17(1):87–103, 2017.
Bobbin, M. L., and J. J. Rossi. RNA Interference (RNAi)-Based Therapeutics: delivering on the Promise? Annu. Rev. Pharmacol. Toxicol. 56:103–122, 2016.
Brody, J. D., W. Z. Ai, D. K. Czerwinski, J. A. Torchia, M. Levy, R. H. Advani, Y. H. Kim, R. T. Hoppe, S. J. Knox, L. K. Shin, I. Wapnir, R. J. Tibshirani, and R. Levy. In situ vaccination with a TLR9 agonist induces systemic lymphoma regression: a phase I/II study. J. Clin. Oncol. 28(28):4324–4332, 2010.
Broz, P., and D. M. Monack. Newly described pattern recognition receptors team up against intracellular pathogens. Nat. Rev. Immunol. 13(8):551–565, 2013.
Brudno, Y., and D. J. Mooney. On-demand drug delivery from local depots. J. Control. Release 219:8–17, 2015.
Chang, E., A. J. McClellan, W. J. Farley, D. Q. Li, S. C. Pflugfelder, and C. S. De Paiva. Biodegradable PLGA-based drug delivery systems for modulating ocular surface disease under experimental murine dry eye. J. Clin. Exp. Ophthalmol. 2(11):191, 2011.
Chen, Q., C. Wang, X. Zhang, G. Chen, Q. Hu, H. Li, J. Wang, D. Wen, Y. Zhang, Y. Lu, G. Yang, C. Jiang, J. Wang, G. Dotti, and Z. Gu. In situ sprayed bioresponsive immunotherapeutic gel for post-surgical cancer treatment. Nat. Nanotechnol. 14(1):89–97, 2019.
Cohen, H., R. J. Levy, J. Gao, I. Fishbein, V. Kousaev, S. Sosnowski, S. Slomkowski, and G. Golomb. Sustained delivery and expression of DNA encapsulated in polymeric nanoparticles. Gene Ther. 7(22):1896–1905, 2000.
Convertine, A. J., D. S. Benoit, C. L. Duvall, A. S. Hoffman, and P. S. Stayton. Development of a novel endosomolytic diblock copolymer for siRNA delivery. J. Control. Release 133(3):221–229, 2009.
Convertine, A. J., C. Diab, M. Prieve, A. Paschal, A. S. Hoffman, P. H. Johnson, and P. S. Stayton. pH-Responsive polymeric micelle carriers for siRNA drugs. Biomacromolecules 11(11):2904–2910, 2010.
Cooper, C., and D. Mackie. Hepatitis B surface antigen-1018 ISS adjuvant-containing vaccine: a review of HEPLISAV safety and efficacy. Expert Rev. Vaccines 10(4):417–427, 2011.
Cun, D., C. Foged, M. Yang, S. Frokjaer, and H. M. Nielsen. Preparation and characterization of poly(d,l-lactide-co-glycolide) nanoparticles for siRNA delivery. Int. J. Pharm. 390(1):70–75, 2010.
Cun, D., D. K. Jensen, M. J. Maltesen, M. Bunker, P. Whiteside, D. Scurr, C. Foged, and H. M. Nielsen. High loading efficiency and sustained release of siRNA encapsulated in PLGA nanoparticles: quality by design optimization and characterization. Eur. J. Pharm. Biopharm. 77(1):26–35, 2011.
Danhier, F., E. Ansorena, J. M. Silva, R. Coco, A. Le Breton, and V. Preat. PLGA-based nanoparticles: an overview of biomedical applications. J. Control. Release 161(2):505–522, 2012.
Elion, D. L., M. E. Jacobson, D. J. Hicks, B. Rahman, V. Sanchez, P. I. Gonzales-Ericsson, O. Fedorova, A. M. Pyle, J. T. Wilson, and R. S. Cook. Therapeutically active RIG-I agonist induces immunogenic tumor cell killing in breast cancers. Cancer Res. 78(21):6183–6195, 2018.
Ferritto, M. S., and D. A. Tirrell. Photoregulation of the binding of an azobenzene-modified poly(methacrylic acid) to phosphatidylcholine bilayer membranes. Biomaterials 11(9):645–651, 1990.
Fire, A., S. Xu, M. K. Montgomery, S. A. Kostas, S. E. Driver, and C. C. Mello. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391(6669):806–811, 1998.
Frauke Pistel, K., A. Breitenbach, R. Zange-Volland, and T. Kissel. Brush-like branched biodegradable polyesters, part III. Protein release from microspheres of poly(vinyl alcohol)-graft-poly(d,l-lactic-co-glycolic acid). J. Control. Release 73(1):7–20, 2001.
Hammerich, L., A. Binder, and J. D. Brody. In situ vaccination: cancer immunotherapy both personalized and off-the-shelf. Mol Oncol 9(10):1966–1981, 2015.
Han, F. Y., K. J. Thurecht, A. K. Whittaker, and M. T. Smith. Bioerodable PLGA-based microparticles for producing sustained-release drug formulations and strategies for improving drug loading. Front Pharmacol. 7:185, 2016.
Ishihara, J., K. Fukunaga, A. Ishihara, H. M. Larsson, L. Potin, P. Hosseinchi, G. Galliverti, M. A. Swartz, and J. A. Hubbell. Matrix-binding checkpoint immunotherapies enhance antitumor efficacy and reduce adverse events. Sci. Transl. Med. 9(415):eaan0401, 2017.
Jacobson, M. E., L. Wang-Bishop, K. W. Becker, and J. T. Wilson. Delivery of 5′-triphosphate RNA with endosomolytic nanoparticles potently activates RIG-I to improve cancer immunotherapy. Biomater. Sci. 7(2):547–559, 2019.
Jewell, C. M., S. C. B. López, and D. J. Irvine. In situ engineering of the lymph node microenvironment via intranodal injection of adjuvant-releasing polymer particles. Proc. Natl. Acad. Sci. USA 108(38):15745–15750, 2011.
Jiang, W., F. G. Zhu, L. Bhagat, D. Yu, J. X. Tang, E. R. Kandimalla, N. La Monica, and S. Agrawal. A toll-like receptor 7, 8, and 9 antagonist inhibits Th1 and Th17 responses and inflammasome activation in a model of IL-23-induced psoriasis. J. Invest. Dermatol. 133(7):1777–1784, 2013.
Johannes, L., and M. Lucchino. Current challenges in delivery and cytosolic translocation of therapeutic RNAs. Nucleic Acid Ther. 28(3):178–193, 2018.
Khan, A., M. Benboubetra, P. Z. Sayyed, K. W. Ng, S. Fox, G. Beck, I. F. Benter, and S. Akhtar. Sustained polymeric delivery of gene silencing antisense ODNs, siRNA, DNAzymes and ribozymes: in vitro and in vivo studies. J. Drug Target. 12(6):393–404, 2004.
Krhac Levacic, A., S. Morys, and E. Wagner. Solid-phase supported design of carriers for therapeutic nucleic acid delivery. Biosci. Rep. 37(5):BSR20160617, 2017.
Kwong, B., S. A. Gai, J. Elkhader, K. D. Wittrup, and D. J. Irvine. Localized immunotherapy via liposome-anchored Anti-CD137 + IL-2 prevents lethal toxicity and elicits local and systemic antitumor immunity. Can. Res. 73(5):1547–1558, 2013.
Kwong, B., H. Liu, and D. J. Irvine. Induction of potent anti-tumor responses while eliminating systemic side effects via liposome-anchored combinatorial immunotherapy. Biomaterials 32(22):5134–5147, 2011.
Langer, R. Drug delivery and targeting. Nature 392(6679 Suppl):5–10, 1998.
Langer, R., and D. A. Tirrell. Designing materials for biology and medicine. Nature 428(6982):487–492, 2004.
Luby, T. M., G. Cole, L. Baker, J. S. Kornher, U. Ramstedt, and M. L. Hedley. Repeated immunization with plasmid DNA formulated in poly(lactide-co-glycolide) microparticles is well tolerated and stimulates durable T cell responses to the tumor-associated antigen cytochrome P450 1B1. Clin. Immunol. 112(1):45–53, 2004.
Lurescia, S., D. Fioretti, and M. Rinaldi. Targeting cytosolic nucleic acid-sensing pathways for cancer immunotherapies. Front. Immunol. 9:711, 2018.
Lurescia, S., D. Fioretti, and M. Rinaldi. Nucleic acid sensing machinery: targeting innate immune system for cancer therapy. Recent Pat. Anticancer Drug Discov. 13(1):2–17, 2018.
Luten, J., C. F. van Nostrum, S. C. De Smedt, and W. E. Hennink. Biodegradable polymers as non-viral carriers for plasmid DNA delivery. J. Control. Release 126(2):97–110, 2008.
Makadia, H. K., and S. J. Siegel. Poly lactic-co-glycolic acid (PLGA) as biodegradable controlled drug delivery carrier. Polymers (Basel) 3(3):1377–1397, 2011.
Malcolm, D. W., M. A. T. Freeberg, Y. Wang, K. R. Sims, H. A. Awad, and D. S. W. Benoit. Diblock copolymer hydrophobicity facilitates efficient gene silencing and cytocompatible nanoparticle-mediated siRNA delivery to musculoskeletal cell types. Biomacromolecules 18(11):3753–3765, 2017.
Mao, S., J. Xu, C. Cai, O. Germershaus, A. Schaper, and T. Kissel. Effect of WOW process parameters on morphology and burst release of FITC-dextran loaded PLGA microspheres. Int. J. Pharm. 334(1–2):137–148, 2007.
Marabelle, A., H. Kohrt, C. Caux, and R. Levy. Intratumoral immunization: a new paradigm for cancer therapy. Clin. Cancer Res. 20(7):1747–1756, 2014.
Marabelle, A., L. Tselikas, T. de Baere, and R. Houot. Intratumoral immunotherapy: using the tumor as the remedy. Ann. Oncol. 28(suppl 12):xii33–xii43, 2017.
Martin, J. R., C. E. Nelson, M. K. Gupta, F. Yu, S. M. Sarett, K. M. Hocking, A. C. Pollins, L. B. Nanney, J. M. Davidson, S. A. Guelcher, and C. L. Duvall. Local delivery of PHD2 siRNA from ROS-degradable scaffolds to promote diabetic wound healing. Adv. Healthc. Mater. 5(21):2751–2757, 2016.
McGinity, J. W., and P. B. O’Donnell. Preparation of microspheres by the solvent evaporation technique. Adv. Drug Deliv. Rev. 28(1):25–42, 1997.
Milling, L., Y. Zhang, and D. J. Irvine. Delivering safer immunotherapies for cancer. Adv. Drug Deliv. Rev. 114:79–101, 2017.
National Center for Immunization and Respiratory Diseases. General recommendations on immunization: recommendations of the Advisory Committee on Immunization Practices (ACIP). MMWR Recomm. Rep. 60(2):1–64, 2011.
Nelson, C. E., M. K. Gupta, E. J. Adolph, J. M. Shannon, S. A. Guelcher, and C. L. Duvall. Sustained local delivery of siRNA from an injectable scaffold. Biomaterials 33(4):1154–1161, 2012.
Nishikawa, M., Y. Mizuno, K. Mohri, N. Matsuoka, S. Rattanakiat, Y. Takahashi, H. Funabashi, D. Luo, and Y. Takakura. Biodegradable CpG DNA hydrogels for sustained delivery of doxorubicin and immunostimulatory signals in tumor-bearing mice. Biomaterials 32(2):488–494, 2011.
Ogawa, Y., M. Yamamoto, H. Okada, T. Yashiki, and T. Shimamoto. A new technique to efficiently entrap leuprolide acetate into microcapsules of polylactic acid or copoly(lactic/glycolic) acid. Chem. Pharm. Bull. (Tokyo) 36(3):1095–1103, 1988.
Ozcan, G., B. Ozpolat, R. L. Coleman, A. K. Sood, and G. Lopez-Berestein. Preclinical and clinical development of siRNA-based therapeutics. Adv. Drug Deliv. Rev. 87:108–119, 2015.
Pannier, A. K., and L. D. Shea. Controlled release systems for DNA delivery. Mol. Ther. 10(1):19–26, 2004.
Pantazis, P., K. Dimas, J. H. Wyche, S. Anant, C. W. Houchen, J. Panyam, and R. P. Ramanujam. Preparation of siRNA-encapsulated PLGA nanoparticles for sustained release of siRNA and evaluation of encapsulation efficiency. Methods Mol. Biol. 906:311–319, 2012.
Park, C. G., C. A. Hartl, D. Schmid, E. M. Carmona, H. J. Kim, and M. S. Goldberg. Extended release of perioperative immunotherapy prevents tumor recurrence and eliminates metastases. Sci. Transl. Med. 10(433):eear1916, 2018.
Patil, Y., and J. Panyam. Polymeric nanoparticles for siRNA delivery and gene silencing. Int. J. Pharm. 367(1–2):195–203, 2009.
Poeck, H., R. Besch, C. Maihoefer, M. Renn, D. Tormo, S. S. Morskaya, S. Kirschnek, E. Gaffal, J. Landsberg, J. Hellmuth, A. Schmidt, D. Anz, M. Bscheider, T. Schwerd, C. Berking, C. Bourquin, U. Kalinke, E. Kremmer, H. Kato, S. Akira, R. Meyers, G. Häcker, M. Neuenhahn, D. Busch, J. Ruland, S. Rothenfusser, M. Prinz, V. Hornung, S. Endres, T. Tüting, and G. Hartmann. 5′-Triphosphate-siRNA: turning gene silencing and RIG-I activation against melanoma. Nat. Med. 14(11):1256–1263, 2008.
Radovic-Moreno, A. F., N. Chernyak, C. C. Mader, S. Nallagatla, R. S. Kang, L. Hao, D. A. Walker, T. L. Halo, T. J. Merkel, C. H. Rische, S. Anantatmula, M. Burkhart, C. A. Mirkin, and S. M. Gryaznov. Immunomodulatory spherical nucleic acids. Proc. Natl. Acad. Sci. USA 112(13):3892–3897, 2015.
Rathbone, M. J., J. Hadgraft, and M. S. Roberts. Modified-release drug delivery technology. London: Taylor & Francis, 2002.
Rothschilds, A. M., and K. D. Wittrup. What, why, where, and when: bringing timing to immuno-oncology. Trends Immunol. 40(1):12–21, 2019.
Sarett, S. M., C. E. Nelson, and C. L. Duvall. Technologies for controlled, local delivery of siRNA. J. Control. Release 218:94–113, 2015.
Senti, G., A. U. Freiburghaus, D. Larenas-Linnemann, H. J. Hoffmann, A. M. Patterson, L. Klimek, D. Di Bona, O. Pfaar, L. Ahlbeck, M. Akdis, D. Weinfeld, F. A. Contreras-Verduzco, A. Pedroza-Melendez, S. H. Skaarup, S. M. Lee, L. O. Cardell, J. M. Schmid, U. Westin, R. Dollner, and T. M. Kundig. Intralymphatic immunotherapy: update and unmet needs. Int. Arch. Allergy Immunol. 178(2):141–149, 2019.
Sioud, M. RNA interference: mechanisms, technical challenges, and therapeutic opportunities. Methods Mol. Biol. 1218:1–15, 2015.
Smith, S. A., L. I. Selby, A. P. R. Johnston, and G. K. Such. The endosomal escape of nanoparticles: towards more efficient cellular delivery. Bioconjug. Chem. 30(2):263–272, 2018.
van den Boorn, J. G., W. Barchet, and G. Hartmann. Nucleic acid adjuvants: toward an educated vaccine. Adv. Immunol. 114:1–32, 2012.
van den Boorn, J. G., and G. Hartmann. Turning tumors into vaccines: co-opting the innate immune system. Immunity 39(1):27–37, 2013.
Wang, L. L., and J. A. Burdick. Engineered hydrogels for local and sustained delivery of RNA-interference therapies. Adv. Healthc. Mater. 6(1):1601041, 2017.
Wang, Y., D. W. Malcolm, and D. S. W. Benoit. Controlled and sustained delivery of siRNA/NPs from hydrogels expedites bone fracture healing. Biomaterials 139:127–138, 2017.
Wang, D., D. R. Robinson, G. S. Kwon, and J. Samuel. Encapsulation of plasmid DNA in biodegradable poly(d,l-lactic-co-glycolic acid) microspheres as a novel approach for immunogene delivery. J. Control. Release 57(1):9–18, 1999.
Wang, C., W. Sun, G. Wright, A. Z. Wang, and Z. Gu. Inflammation-triggered cancer immunotherapy by programmed delivery of CpG and anti-PD1 antibody. Adv. Mater. 28(40):8912–8920, 2016.
Whitehead, K. A., R. Langer, and D. G. Anderson. Knocking down barriers: advances in siRNA delivery. Nat. Rev. Drug Discov. 8(2):129–138, 2009.
Wilson, J. T., S. Keller, M. J. Manganiello, C. Cheng, C.-C. Lee, C. Opara, A. Convertine, and P. S. Stayton. pH-Responsive nanoparticle vaccines for dual-delivery of antigens and immunostimulatory oligonucleotides. ACS Nano. 7(5):3912–3925, 2013.
Woodrow, K. A., Y. Cu, C. J. Booth, J. K. Saucier-Sawyer, M. J. Wood, and W. M. Saltzman. Intravaginal gene silencing using biodegradable polymer nanoparticles densely loaded with small-interfering RNA. Nat. Mater. 8(6):526–533, 2009.
Wu, S. Y., G. Lopez-Berestein, G. A. Calin, and A. K. Sood. RNAi therapies: drugging the undruggable. Sci. Transl. Med. 6(240):240, 2014.
Wu-Pong, S., and Y. Rojanasakul. Biopharmaceutical Drug Design and Development (2nd ed.). Totowa: Humana Press, p. 375, 2008.
Yan, J., Z.-Y. Wang, H.-Z. Yang, H.-Z. Liu, S. Mi, X.-X. Lv, X.-M. Fu, H.-M. Yan, X.-W. Zhang, Q.-M. Zhan, and Z.-W. Hu. Timing is critical for an effective anti-metastatic immunotherapy: the decisive role of IFNγ/STAT1-mediated activation of autophagy. PLoS ONE 6(9):e24705, 2011.
Young, K. H., J. R. Baird, T. Savage, B. Cottam, D. Friedman, S. Bambina, D. J. Messenheimer, B. Fox, P. Newell, K. S. Bahjat, M. J. Gough, and M. R. Crittenden. Optimizing timing of immunotherapy improves control of tumors by hypofractionated radiation therapy. PLoS ONE 11(6):e0157164, 2016.
Zhang, L., W. Wang, and S. Wang. Effect of vaccine administration modality on immunogenicity and efficacy. Expert Rev. Vaccines 14(11):1509–1523, 2015.
Zhu, F. G., W. Jiang, L. Bhagat, D. Wang, D. Yu, J. X. Tang, E. R. Kandimalla, N. La Monica, and S. Agrawal. A novel antagonist of Toll-like receptors 7, 8 and 9 suppresses lupus disease-associated parameters in NZBW/F1 mice. Autoimmunity 46(7):419–428, 2013.
Zhu, X., F. Nishimura, K. Sasaki, M. Fujita, J. E. Dusak, J. Eguchi, W. Fellows-Mayle, W. J. Storkus, P. R. Walker, A. M. Salazar, and H. Okada. Toll like receptor-3 ligand poly-ICLC promotes the efficacy of peripheral vaccinations with tumor antigen-derived peptide epitopes in murine CNS tumor models. J. Transl. Med. 5:10, 2007.
We gratefully acknowledge Dr. Bob Weinberg and Dr. Didier Trono for gifts of plasmids via Addgene.org. We thank Dr. Steven Goodbred Jr. and his laboratory for use of the Mastersizer 2000 (Malvern, USA). We thank Kyle Becker for his assistance with the orthotopic tumor inoculations. We thank the core facilities of Vanderbilt, including the Vanderbilt Institute of Nanoscale Sciences and Engineering (VINSE) for the use of both the Zetasizer Nano ZS Instrument (Malvern, USA) and the Zeiss Merlin SEM (Carl Zeiss Microscopy, LLC, ZEISS Group, Thornwood, NY), the Vanderbilt Translational Pathology Shared Resource (supported in part by the NCI/NIH Cancer Center Support Grant 5P30 CA684850-19) for cryosectioning of tumor samples, and Vanderbilt University Medical Center Flow Cytometry Shared Resource (supported by the Vanderbilt Ingram Cancer Center P30 CA68485) and the Vanderbilt Digestive Disease Research Center (DK058404) for cell sorting. This research was supported by grants from Alex’s Lemonade Stand Foundation ‘A’ Award SID924 (JTW) and Pediatric Oncology Student Training (POST) Award cosponsored by Love Your Mellon (KMG), the American Cancer Society Institutional Research Grant IRG-58-009-56 (JTW), the Congressionally-Directed Medical Research Program W81XWH-161-0063 (JTW) and W81XWH-161-0063 (RSC), the National Institutes of Health R01CA224241 (CLD) and R01EB019409 (CLD), and the National Science Foundation Graduate Research Fellowship Program 0909667 and 1445197 (KVK).
Conflict of interest
The authors declare no conflicts of interest.
All animal experiments were approved by the Vanderbilt University Institutional Animal Care and Use Committee (IACUC), and all surgical and experimental procedures were performed in accordance with the regulations and guidelines of the Vanderbilt University IACUC. Female BALB/cJ mice (6–8 weeks old; The Jackson Laboratory, Bar Harbor, ME) were maintained at the animal facilities of Vanderbilt University under specific pathogen-free conditions. Tumor volume, total mass, and animal well-being were monitored every other day.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
John T. Wilson is an Assistant Professor of Chemical and Biomolecular Engineering and Biomedical Engineering at Vanderbilt University. He received his B.S. in Bioengineering from Oregon State University and his Ph.D. from the Georgia Institute of Technology in the laboratory of Professor Elliot L. Chaikof, where he was awarded a Whitaker Foundation Graduate Fellowship. He then joined the laboratory of Professor Patrick Stayton at the University of Washington with support of a Cancer Research Institute Postdoctoral Fellowship. He started his independent laboratory at Vanderbilt in January of 2014, where his group works at the interface of molecular engineering and immunology to innovate technologies to improve human health. His multidisciplinary research program is supported by productive and synergistic collaborations with oncologists, cancer biologists, immunologists, chemists, and other engineers. Since establishing his lab at Vanderbilt, he has been awarded the NSF CAREER Award, an ‘A’ Award from Alex’s Lemonade Stand Foundation, a Melanoma Research Alliance Young Investigator Award, an Innovative Research Grant from Stand Up To Cancer, and has been named an Emerging Investigator by Biomaterials Science.
This article is part of the 2019 CMBE Young Innovators special issue.
Associate Editor Michael R. King oversaw the review of this article.
About this article
Cite this article
Garland, K.M., Sevimli, S., Kilchrist, K.V. et al. Microparticle Depots for Controlled and Sustained Release of Endosomolytic Nanoparticles. Cel. Mol. Bioeng. 12, 429–442 (2019). https://doi.org/10.1007/s12195-019-00571-6
- Nucleic acid therapeutics
- Local delivery
- RNA interference
- Endosomal escape
- Drug delivery depot