title: Non-virally engineered human adipose mesenchymal stem cells produce BMP4, target brain tumors, and extend survival.
abstract: There is a need for enabling non-viral nanobiotechnology to allow safe and effective gene therapy and cell therapy, which can be utilized to treat devastating diseases such as brain cancer. Human adipose-derived mesenchymal stem cells (hAMSCs) display high anti-glioma tropism and represent a promising delivery vehicle for targeted brain tumor therapy. In this study, we demonstrate that non-viral, biodegradable polymeric nanoparticles (NPs) can be used to engineer hAMSCs with higher efficacy (75% of cells) than leading commercially available reagents and high cell viability. To accomplish this, we engineered a poly(beta-amino ester) (PBAE) polymer structure to transfect hAMSCs with significantly higher efficacy than Lipofectamine™ 2000. We then assessed the ability of NP-engineered hAMSCs to deliver bone morphogenetic protein 4 (BMP4), which has been shown to have a novel therapeutic effect by targeting human brain tumor initiating cells (BTIC), a source of cancer recurrence, in a human primary malignant glioma model. We demonstrated that hAMSCs genetically engineered with polymeric nanoparticles containing BMP4 plasmid DNA (BMP4/NP-hAMSCs) secrete BMP4 growth factor while maintaining their multipotency and preserving their migration and invasion capacities. We also showed that this approach can overcome a central challenge for brain therapeutics, overcoming the blood brain barrier, by demonstrating that NP-engineered hAMSCs can migrate to the brain and penetrate the brain tumor after both intranasal and systemic intravenous administration. Critically, athymic rats bearing human primary BTIC-derived tumors and treated intranasally with BMP4/NP-hAMSCs showed significantly improved survival compared to those treated with control GFP/NP-hAMCSs. This study demonstrates that synthetic polymeric nanoparticles are a safe and effective approach for stem cell-based cancer-targeting therapies.
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Antonella
authors.initials: A
authors.lastname: Mangraviti
authors.affiliation: Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD, USA.
authors.forename: Stephany Y
authors.initials: SY
authors.lastname: Tzeng
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: David
authors.initials: D
authors.lastname: Gullotti
authors.affiliation: Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD, USA.
authors.forename: Kristen L
authors.initials: KL
authors.lastname: Kozielski
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Kim
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Seng
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Sara
authors.initials: S
authors.lastname: Abbadi
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Paula
authors.initials: P
authors.lastname: Schiapparelli
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Sarabia-Estrada
authors.affiliation: IRCSS Casa Sollievo della Sofferenza, Opera di San Pio da Pietrelcina, viale dei Cappuccini, 1, 71013 S. Giovanni Rotondo, Italy.
authors.forename: Angelo
authors.initials: A
authors.lastname: Vescovi
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Henry
authors.initials: H
authors.lastname: Brem
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Olivi
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: btyler@jhmi.edu.
authors.forename: Betty
authors.initials: B
authors.lastname: Tyler
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Biomedical Engineering, Johns Hopkins School of Medicine, Baltimore, MD, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: green@jhu.edu.
authors.forename: Jordan J
authors.initials: JJ
authors.lastname: Green
authors.affiliation: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: aquinon2@jhmi.edu.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Quinones-Hinojosa
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics*; D001932:Brain Neoplasms / Q000628:therapy*; D002460:Cell Line; D045744:Cell Line, Tumor; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D005260:Female; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D051381:Rats; D011923:Rats, Nude; D014162:Transfection / Q000379:methods
chemical_list: C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; D011108:Polymers; C507253:poly(beta-amino ester); D004247:DNA
keywords: Adipose-derived stem cells; Brain cancer; Gene delivery; Nanoparticles; Tumor stem cells