{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T17:49:26Z","timestamp":1766598566479,"version":"build-2065373602"},"reference-count":46,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T00:00:00Z","timestamp":1627257600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["2016M3A9B5942352"],"award-info":[{"award-number":["2016M3A9B5942352"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Hanyang University General Research Grant","award":["HY-2011-G-201100000001880"],"award-info":[{"award-number":["HY-2011-G-201100000001880"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Adenovirus (Ad) has risen to be a promising alternative to conventional cancer therapy. However, systemic delivery of Ad, which is necessary for the treatment of metastatic cancer, remains a major challenge within the field, owing to poor tumor tropism and nonspecific hepatic tropism of the virus. To address this limitation of Ad, we have synthesized two variants of folic acid (FA)-conjugated methoxy poly(ethylene glycol)-b-poly{N-[N-(2-aminoethyl)-2-aminoethyl]-L-glutamate (P5N2LG-FA and P5N5LG-FA) using 5 kDa poly(ethylene glycol) (PEG) with a different level of protonation (N2 &lt; N5 in terms of charge), along with a P5N5LG control polymer without FA. Our findings demonstrate that P5N5LG, P5N2LG-FA, and P5N5LG-FA exert a lower level of cytotoxicity compared to 25 kDa polyethyleneimine. Furthermore, green fluorescent protein (GFP)-expressing Ad complexed with P5N2LG-FA and P5N5LG-FA (Ad\/P5N2LG-FA and Ad\/P5N5LG-FA, respectively) exerted superior transduction efficiency compared to naked Ad or Ad complexed with P5N5LG (Ad\/P5N5LG) in folate receptor (FR)-overexpressing cancer cells (KB and MCF7). All three nanocomplexes (Ad\/P5N5LG, Ad\/P5N2LG-FA, and Ad\/P5N5LG-FA) internalized into cancer cells through coxsackie adenovirus receptor-independent endocytic mechanism and the cell uptake was more efficient than naked Ad. Importantly, the cell uptake of the two FA functionalized nanocomplexes (Ad\/P5N2LG-FA and Ad\/P5N5LG-FA) was dependent on the complementary interaction of FA\u2013FR. Systemically administered Ad\/P5N5LG, Ad\/P5N2LG-FA, and Ad\/P5N5LG-FA showed exponentially higher retainment of the virus in blood circulation up to 24 h post-administration compared with naked Ad. Both tumor-targeted nanocomplexes (Ad\/P5N2LG-FA and Ad\/P5N5LG-FA) showed significantly higher intratumoral accumulation than naked Ad or Ad\/P5N5LG via systemic administration. Both tumor-targeted nanocomplexes accumulated at a lower level in liver tissues compared to naked Ad. Notably, the nonspecific accumulation of Ad\/P5N2LG-FA was significantly lower than Ad\/P5N5LG-FA in several normal organs, while exhibiting a significantly higher intratumoral accumulation level, showing that careful optimization of polyplex surface charge is critical to successful tumor-targeted systemic delivery of Ad nanocomplexes.<\/jats:p>","DOI":"10.3390\/cells10081896","type":"journal-article","created":{"date-parts":[[2021,7,26]],"date-time":"2021-07-26T22:22:46Z","timestamp":1627338166000},"page":"1896","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Optimizing Active Tumor Targeting Biocompatible Polymers for Efficient Systemic Delivery of Adenovirus"],"prefix":"10.3390","volume":"10","author":[{"given":"Jun Young","family":"Lee","sequence":"first","affiliation":[{"name":"Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jin Woo","family":"Hong","sequence":"additional","affiliation":[{"name":"GeneMedicine Co., Ltd., 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thavasyappan","family":"Thambi","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A-Rum","family":"Yoon","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, Korea"},{"name":"Institute of Nano Science and Technology (INST), Hanyang University, Seoul 04763, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joung-Woo","family":"Choi","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yi","family":"Li","sequence":"additional","affiliation":[{"name":"Theranostic Macromolecules Research Center, School of Chemical Engineering, Sungkyunkwan University, Suwon 16419, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Quang Nam","family":"Bui","sequence":"additional","affiliation":[{"name":"Theranostic Macromolecules Research Center, School of Chemical Engineering, Sungkyunkwan University, Suwon 16419, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Doo Sung","family":"Lee","sequence":"additional","affiliation":[{"name":"Theranostic Macromolecules Research Center, School of Chemical Engineering, Sungkyunkwan University, Suwon 16419, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9466-4531","authenticated-orcid":false,"given":"Chae-Ok","family":"Yun","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, College of Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, Korea"},{"name":"GeneMedicine Co., Ltd., 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, Korea"},{"name":"Institute of Nano Science and Technology (INST), Hanyang University, Seoul 04763, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,7,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1016\/j.addr.2011.12.011","article-title":"Evolution of oncolytic adenovirus for cancer treatment","volume":"64","author":"Choi","year":"2012","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1016\/j.tibtech.2009.01.003","article-title":"Designer adenoviruses for nanomedicine and nanodiagnostics","volume":"27","author":"Singh","year":"2009","journal-title":"Trends Biotechnol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1021\/mp050045c","article-title":"Genetic targeting strategies for adenovirus","volume":"2","author":"Noureddini","year":"2005","journal-title":"Mol. Pharm."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/j.jconrel.2011.06.014","article-title":"Viral genome DNA\/lipoplexes elicit in situ oncolytic viral replication and potent antitumor efficacy via systemic delivery","volume":"155","author":"Kwon","year":"2011","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1016\/j.jconrel.2015.10.009","article-title":"Polymeric oncolytic adenovirus for cancer gene therapy","volume":"219","author":"Choi","year":"2015","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1016\/j.jconrel.2011.06.001","article-title":"Targeted drug delivery to tumors: Myths, reality and possibility","volume":"153","author":"Bae","year":"2011","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"479","DOI":"10.1016\/S1359-6446(02)02243-2","article-title":"Targeting tumors with non-viral gene delivery systems","volume":"7","author":"Ogris","year":"2002","journal-title":"Drug Discov. Today"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1615","DOI":"10.1016\/j.addr.2008.08.005","article-title":"Active targeting schemes for nanoparticle systems in cancer therapeutics","volume":"60","author":"Byrne","year":"2008","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_9","first-page":"51","article-title":"Antibody conjugation methods for active targeting of liposomes","volume":"25","author":"Ansell","year":"2000","journal-title":"Methods Mol. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"322","DOI":"10.7150\/thno.10257","article-title":"Aptamers: Active targeting ligands for cancer diagnosis and therapy","volume":"5","author":"Wu","year":"2015","journal-title":"Theranostics"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"329","DOI":"10.3390\/pharmaceutics5020329","article-title":"Polysaccharide-based micelles for drug delivery","volume":"5","author":"Zhang","year":"2013","journal-title":"Pharmaceutics"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"698","DOI":"10.3109\/1061186X.2014.910792","article-title":"Anti-neuropilin 1 antibody Fab\u2019 fragment conjugated liposomal docetaxel for active targeting of tumours","volume":"22","author":"Manjappa","year":"2014","journal-title":"J. Drug Target."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2314","DOI":"10.1016\/j.biomaterials.2010.10.031","article-title":"Active targeting and safety profile of PEG-modified adenovirus conjugated with herceptin","volume":"32","author":"Kim","year":"2011","journal-title":"Biomaterials"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"766","DOI":"10.1016\/j.jconrel.2015.10.015","article-title":"Potent antitumor effect of neurotensin receptor-targeted oncolytic adenovirus co-expressing decorin and Wnt antagonist in an orthotopic pancreatic tumor model","volume":"220","author":"Na","year":"2015","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.jconrel.2016.02.046","article-title":"Antitumor effect and safety profile of systemically delivered oncolytic adenovirus complexed with EGFR-targeted PAMAM-based dendrimer in orthotopic lung tumor model","volume":"231","author":"Yoon","year":"2016","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.jconrel.2010.08.027","article-title":"To exploit the tumor microenvironment: Passive and active tumor targeting of nanocarriers for anti-cancer drug delivery","volume":"148","author":"Danhier","year":"2010","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.ijpharm.2005.10.010","article-title":"Opsonization, biodistribution, and pharmacokinetics of polymeric nanoparticles","volume":"307","author":"Owens","year":"2006","journal-title":"Int. J. Pharm."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1042\/bj20031253","article-title":"Size-dependent internalization of particles via the pathways of clathrin- and caveolae-mediated endocytosis","volume":"377","author":"Rejman","year":"2004","journal-title":"Biochem. J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4573","DOI":"10.1016\/S0142-9612(02)00203-X","article-title":"Receptor-independent augmentation of adenovirus-mediated gene transfer with chitosan in vitro","volume":"23","author":"Kawamata","year":"2002","journal-title":"Biomaterials"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Wang, I.J., Jhuang, M.C., Chen, Y.H., Yeh, L.K., Liu, C.Y., and Young, T.H. (2010). Chitosan modification of adenovirus to modify transfection efficiency in bovine corneal epithelial cells. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0012085"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1021\/bm501116x","article-title":"Safety profiles and antitumor efficacy of oncolytic adenovirus coated with bioreducible polymer in the treatment of a CAR negative tumor model","volume":"16","author":"Jung","year":"2015","journal-title":"Biomacromolecules"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1865","DOI":"10.1016\/j.biomaterials.2009.11.043","article-title":"The effect of surface modification of adenovirus with an arginine-grafted bioreducible polymer on transduction efficiency and immunogenicity in cancer gene therapy","volume":"31","author":"Kim","year":"2010","journal-title":"Biomaterials"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1016\/j.jconrel.2015.09.068","article-title":"Hepatoma targeting peptide conjugated bio-reducible polymer complexed with oncolytic adenovirus for cancer gene therapy","volume":"220","author":"Choi","year":"2015","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.biomaterials.2017.08.035","article-title":"Synergistic antitumor effect mediated by a paclitaxel-conjugated polymeric micelle-coated oncolytic adenovirus","volume":"145","author":"Kasala","year":"2017","journal-title":"Biomaterials"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1016\/j.jconrel.2010.11.004","article-title":"Endosomal escape pathways for delivery of biologicals","volume":"151","author":"Varkouhi","year":"2011","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1689","DOI":"10.2217\/nnm-2016-0060","article-title":"Evolving lessons on nanomaterial-coated viral vectors for local and systemic gene therapy","volume":"11","author":"Kasala","year":"2016","journal-title":"Nanomedicine (Lond. Engl.)"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1517\/17425247.2016.1158707","article-title":"Redirecting adenovirus tropism by genetic, chemical, and mechanical modification of the adenovirus surface for cancer gene therapy","volume":"13","author":"Yoon","year":"2016","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"897","DOI":"10.1021\/bc049951i","article-title":"Toxicity of gold nanoparticles functionalized with cationic and anionic side chains","volume":"15","author":"Goodman","year":"2004","journal-title":"Bioconjug. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1002\/smll.200901158","article-title":"Effect of surface properties on nanoparticle-cell interactions","volume":"6","author":"Verma","year":"2010","journal-title":"Small (Weinh. Der Bergstr. Ger.)"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.biomaterials.2014.11.021","article-title":"Dual tumor targeting with pH-sensitive and bioreducible polymer-complexed oncolytic adenovirus","volume":"41","author":"Moon","year":"2015","journal-title":"Biomaterials"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/S0169-409X(99)00062-9","article-title":"Targeted drug delivery via the folate receptor","volume":"41","author":"Sudimack","year":"2000","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.jconrel.2015.01.005","article-title":"pH-sensitive oncolytic adenovirus hybrid targeting acidic tumor microenvironment and angiogenesis","volume":"205","author":"Choi","year":"2015","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/j.jconrel.2013.03.030","article-title":"Therapeutic targeting of chitosan-PEG-folate-complexed oncolytic adenovirus for active and systemic cancer gene therapy","volume":"169","author":"Kwon","year":"2013","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1016\/j.jconrel.2011.12.008","article-title":"Linearized oncolytic adenoviral plasmid DNA delivered by bioreducible polymers","volume":"158","author":"Kim","year":"2012","journal-title":"J. Control. Release Off. J. Control. Release Soc."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"1109","DOI":"10.1038\/mt.2013.100","article-title":"FX and host defense evasion tactics by adenovirus","volume":"21","author":"Baker","year":"2013","journal-title":"Mol. Ther. J. Am. Soc. Gene Ther."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1039\/C7BM00128B","article-title":"Bioresorbable polypeptide-based comb-polymers efficiently improves the stability and pharmacokinetics of proteins in vivo","volume":"5","author":"Turabee","year":"2017","journal-title":"Biomater. Sci."},{"key":"ref_37","first-page":"5577","article-title":"The role of surface charge in cellular uptake and cytotoxicity of medical nanoparticles","volume":"7","year":"2012","journal-title":"Int. J. Nanomed."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1818","DOI":"10.1021\/acs.bioconjchem.5b00357","article-title":"Tuning Surface Charge and PEGylation of Biocompatible Polymers for Efficient Delivery of Nucleic Acid or Adenoviral Vector","volume":"26","author":"Choi","year":"2015","journal-title":"Bioconjug. Chem."},{"key":"ref_39","first-page":"6831","article-title":"Cationic charge determines the distribution of liposomes between the vascular and extravascular compartments of tumors","volume":"62","author":"Campbell","year":"2002","journal-title":"Cancer Res."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Hong, J., and Yun, C.O. (2019). Overcoming the limitations of locally administered oncolytic virotherapy. BMC Biomed. Eng., 1.","DOI":"10.1186\/s42490-019-0016-x"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1016\/j.addr.2006.03.007","article-title":"Current status of polymeric gene delivery systems","volume":"58","author":"Park","year":"2006","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1002\/jgm.237","article-title":"Side-effects of a systemic injection of linear polyethylenimine-DNA complexes","volume":"4","author":"Chollet","year":"2002","journal-title":"J. Gene Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"5779","DOI":"10.1007\/s00216-017-0527-z","article-title":"Zeta potential: A case study of cationic, anionic, and neutral liposomes","volume":"409","author":"Smith","year":"2017","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Arvizo, R.R., Miranda, O.R., Moyano, D.F., Walden, C.A., Giri, K., Bhattacharya, R., Robertson, J.D., Rotello, V.M., Reid, J.M., and Mukherjee, P. (2011). Modulating pharmacokinetics, tumor uptake and biodistribution by engineered nanoparticles. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0024374"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1038\/nbt.3330","article-title":"Principles of nanoparticle design for overcoming biological barriers to drug delivery","volume":"33","author":"Blanco","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3435","DOI":"10.1016\/j.biomaterials.2011.01.021","article-title":"The effect of surface charge on in vivo biodistribution of PEG-oligocholic acid based micellar nanoparticles","volume":"32","author":"Xiao","year":"2011","journal-title":"Biomaterials"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/10\/8\/1896\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:35:14Z","timestamp":1760164514000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/10\/8\/1896"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,7,26]]},"references-count":46,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2021,8]]}},"alternative-id":["cells10081896"],"URL":"https:\/\/doi.org\/10.3390\/cells10081896","relation":{},"ISSN":["2073-4409"],"issn-type":[{"type":"electronic","value":"2073-4409"}],"subject":[],"published":{"date-parts":[[2021,7,26]]}}}