Abstract:
:Reporter gene-based molecular imaging can provide invaluable information on the fate of cellular therapies postimplantation. Integrating lentiviral vectors (ILVs) are commonly used for stably engineering cells; however, their potential for insertional mutagenesis poses a significant safety concern and barrier to widespread clinical adoption. In cells that slowly divide or are postmitotic in vivo, such as mesenchymal stem cells (MSCs), nonintegrating lentiviral vectors (NILVs) may be a safer alternative option because NILVs remain episomal and can provide prolonged expression profiles. Here, NILVs coexpressing fluorescence and bioluminescence imaging (BLI) reporters were engineered and used to longitudinally image the viability of human MSCs in vitro and in vivo in mice. In vitro, ILV-transduced cancer cells and MSCs maintained steady reporter gene expression over time, whereas NILV-transduced cells progressively lost signal. NILV reporter loss was accelerated in highly proliferative cancer cells compared with less proliferative MSCs. In vivo, ILV- and NILV-transduced MSCs were each detectable with BLI postintramuscular implantation, with significantly higher ILV-based signal compared with NILV-based signal. BLI signal was observed to similarly diminish over time for both cell populations, which was attributed to cell death. Despite the reduced signal intensity with NILVs and the minimal number of cells injected (40,000), live NILV-transduced MSCs were reliably visualized for up to 2 weeks. Safety is a concern for future clinical reporter gene applications. We present NILVs as a safe means of imaging reporter gene expression for slowly dividing or nondividing cells and showed effective tracking of the viability of a small number of live transplanted MSCs over time with optical imaging. Future work will evaluate improvements to episomal NILV reporter expression, explore sensitive clinically relevant reporters, and apply this approach to clinically relevant MSC applications in preclinical models. NILVs may have broad clinical applications for expression of imaging reporters and other gene products in MSC-based therapies.
journal_name
Hum Gene Therjournal_title
Human gene therapyauthors
Hamilton AM,Foster PJ,Ronald JAdoi
10.1089/hum.2018.111subject
Has Abstractpub_date
2018-10-01 00:00:00pages
1213-1225issue
10eissn
1043-0342issn
1557-7422journal_volume
29pub_type
杂志文章abstract::Adenoviral vectors used in gene therapy are predominantly derived from adenovirus serotype 5 (Ad5), which infects a broad range of cells. Ad5 cell entry involves interactions with the coxsackie-adenovirus receptor (CAR) and integrins. To assess these receptors in vivo, we mutated amino acid residues in fiber and pento...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/104303403765255165
更新日期:2003-05-20 00:00:00
abstract::Human bocavirus type-1 (HBoV1) has a high tropism for the apical membrane of human airway epithelia. The packaging of a recombinant adeno-associated virus 2 (rAAV2) genome into HBoV1 capsid produces a chimeric vector (rAAV2/HBoV1) that also efficiently transduces human airway epithelia. As such, this vector is attract...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.2017.060
更新日期:2017-08-01 00:00:00
abstract::Adult T cell leukemia/lymphoma (ATL) is derived from CD4+ T cells and has a poor prognosis because of its resistance to chemotherapy. To evaluate the effectiveness of gene therapy for ATL, the effect of ganciclovir on ATL cell lines transfected with the thymidine kinase gene of herpes simplex virus type 1 (HSV-TK) was...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.1996.7.18-2203
更新日期:1996-12-01 00:00:00
abstract::The enzyme hypoxanthine-guanine phosphoribosyltransferase (HGPRT) expressed by the parasite Trypanosoma brucei (Tb) can convert allopurinol, a purine analogue, to corresponding nucleotides with greater efficiency than its human homologue. We have developed a retroviral system that expresses the parasitic enzyme and te...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/10430340152528165
更新日期:2001-09-01 00:00:00
abstract::Eleven patients with moderate to severe erectile dysfunction (ED) were given a single-dose corpus cavernosum injection of hMaxi-K, a "naked" DNA plasmid carrying the human cDNA encoding hSlo (for human slow-poke), the gene for the alpha, or pore-forming, subunit of the human smooth muscle Maxi-K channel. Three patient...
journal_title:Human gene therapy
pub_type: 杂志文章,多中心研究
doi:10.1089/hum.2006.17.1165
更新日期:2006-12-01 00:00:00
abstract::Advanced prostate cancer is invariably lethal once it becomes androgen independent (AI). With the aim of developing a new treatment we have used the human androgen-independent prostate cancer cell line, PC-3, to evaluate the effectiveness of two enzyme-directed prodrug therapy (EPT) systems as a novel means for promot...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.1998.9.11-1617
更新日期:1998-07-20 00:00:00
abstract::The use of gene-engineered T cells expressing chimeric single-chain (scFv) receptors capable of codelivering CD28 costimulation and T cell receptor zeta chain (TCR-zeta) activation signals has emerged as a promising treatment regimen for cancer. Using retroviral transduction, primary human T lymphocytes were gene-engi...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/1043034041361235
更新日期:2004-07-01 00:00:00
abstract::Current HIV-1 gene therapy approaches aim at stopping the viral life cycle at its earliest steps, such as entry or immediate postentry events. Among the most widely adopted strategies are CCR5 downregulation/knockout and the use of broadly neutralizing antibodies. However, the long-term efficacy and side effects are s...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.2015.059
更新日期:2015-10-01 00:00:00
abstract::Interleukin (IL)-12 has been reported to induce cellular immune responses for protection against tumor formation. Here we investigate the utility of adenoviral delivery of IL-12 as an adjuvant for a human papillomavirus E7 subunit vaccine in a mouse tumor challenge model. Direct intratumoral injection of AdIL-12 resul...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/104303403769211619
更新日期:2003-10-10 00:00:00
abstract::This multicenter phase I/II study evaluated the safety, pharmacokinetics, and antitumor effects of repeated doses of NV1020, a genetically engineered oncolytic herpes simplex virus, in patients with advanced metastatic colorectal cancer (mCRC). Patients with liver-dominant mCRC received four fixed NV1020 doses via wee...
journal_title:Human gene therapy
pub_type: 杂志文章,多中心研究
doi:10.1089/hum.2010.020
更新日期:2010-09-01 00:00:00
abstract::We evaluated the ability of a replication-deficient, recombinant adenoviral vector to transfer the bifunctional gene GAL-TEK, which expresses a marking/therapeutic gene product, to naturally occurring cat fibrosarcomas in situ. GAL-TEK contains an in-frame fusion of the bacterial LacZ gene for histochemical marking of...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.1995.6.9-1215
更新日期:1995-09-01 00:00:00
abstract::Hepatic stellate cells (HSCs) are the primary cell type responsible for liver fibrogenesis. Transforming growth factor beta 1 (TGF-β1) and platelet-derived growth factor (PDGF) are key profibrotic cytokines that regulate HSC activation and proliferation with functional convergence. Dual RNA interference against their ...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.2018.047
更新日期:2019-02-01 00:00:00
abstract::Guidelines for testing gene therapy products for ecotropic replication-competent retrovirus (Eco-RCR) have not been delineated as they have for amphotropic viruses. To evaluate biologic assays that can detect these viruses, we compared an S(+)/L(-) assay and a marker rescue assay designed specifically for Eco-RCR dete...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/104303402760293619
更新日期:2002-09-20 00:00:00
abstract::Accurate quantification of gene transfer (or gene correction) is a universal challenge in the field of gene therapy. In developing a clinical trial of lymphocyte gene therapy for Hunter syndrome (mucopolysaccharidosis type II), methods using Southern blot or automated DNA sequencing technology were employed, but found...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/10430349950016898
更新日期:1999-10-10 00:00:00
abstract::Recombinant adenoviruses have great potential as gene delivery systems because of their ability to infect a wide range of target cells. However, systemic delivery of viral vectors to tissues other than liver and spleen has been inefficient because of the rapid clearance of the circulating virus by the liver. In the pr...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/10430340050015806
更新日期:2000-03-01 00:00:00
abstract::Gyrate atrophy is a progressive blindness associated with deficiency of ornithine aminotransferase (OAT). The strategy of using an autologous keratinocyte graft, modified to express high levels of OAT as an ornithine-catabolizing skin-based enzyme sink, is investigated. Two OAT-containing retroviral vectors were const...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.1997.8.17-2125
更新日期:1997-11-20 00:00:00
abstract::This study was designed to retrovirally transduce T cells by a protocol that would be simple, short, cost effective, applicable for clinical use, and efficient enough to avoid further selection of transduced T cells. Because retrovirally mediated infection is depending on the cell cycle, we first optimized the conditi...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/10430340050015239
更新日期:2000-05-20 00:00:00
abstract::Restoration of correct splicing of βIVS2-654-globin pre-mRNA was previously accomplished in erythroid cells from β-thalassemia/HbE patients by an engineered U7 small nuclear RNA (snRNA) that carried a sequence targeted to the cryptic branch point and an exonic splicing enhancer, U7.BP+623 snRNA. In this study, this ap...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.2020.145
更新日期:2020-11-02 00:00:00
abstract::Human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) offer great hope for in vitro modeling of Parkinson's disease (PD), as well as for designing cell-replacement therapies. To realize these opportunities, there is an urgent need to develop efficient protocols for the directed differentiation of...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.2011.054
更新日期:2012-01-01 00:00:00
abstract::The influence of serum on the production of retroviral vectors by the HT1080 human fibrosarcoma-derived packaging cell line FLYRD18 was investigated. A fourfold increase in virus titer was observed under serum-free conditions, as compared with medium supplemented with 10% fetal calf serum. A similar improvement was al...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/10430349950017329
更新日期:1999-08-10 00:00:00
abstract::Oncolytic measles virus (MV) encoding the human thyroidal sodium iodide symporter (MV-NIS) has proved to be safe after intraperitoneal or intravenous administration in patients with ovarian cancer or multiple myeloma, respectively, but it has not yet been administered through intratumoral injection in humans. Squamous...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.2011.128
更新日期:2012-03-01 00:00:00
abstract::One of the major obstacles to pulmonary-directed gene therapy using adenoviral vectors is the induction of inflammation. We investigated whether the adenoviral particles that constitute the initial inoculum can serve as an inflammatory stimulus, independent of their ability to express genes that they contain. Viral pa...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.1995.6.12-1553
更新日期:1995-12-01 00:00:00
abstract::Clinical applications of gene therapy require advances in gene delivery systems. Although numerous clinical trials are already underway, the ultimate success of gene therapies will depend on gene transfer vectors that facilitate the expression of a specific gene at therapeutic levels in the desired cell populations wi...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/10430340360535751
更新日期:2003-02-10 00:00:00
abstract::Systemic administration of adenoviral vectors leads to activation of innate and antigen-specific immunity. In an attempt to diminish T and B cell-specific immune responses to E1-deleted adenoviral vectors, capsid proteins were modified with various activated monomethoxypolyethylene glycols (MPEGs). The impact of this ...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/104303402760372972
更新日期:2002-10-10 00:00:00
abstract::Replication-deficient adenoviruses are known to induce acute injury and inflammation of infected tissues, thus limiting their use for human gene therapy. However, molecular mechanisms triggering this response have not been fully defined. To characterize this response, chemokine expression was evaluated in DBA/2 mice f...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/10430349950018364
更新日期:1999-04-10 00:00:00
abstract::Neurodegeneration in Parkinson's disease (PD) affects mainly dopaminergic neurons in the substantia nigra, where age-related, increasing percentages of cells lose detectable respiratory activity associated with depletion of intact mitochondrial DNA (mtDNA). Replenishment of mtDNA might improve neuronal bioenergetic fu...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.2009.023
更新日期:2009-08-01 00:00:00
abstract::Adenovirus-polylysine-DNA complexes were evaluated for their capacity to accomplish direct in vivo gene transfer to airway epithelium employing a rodent model. Binary complexes containing transferrin or adenovirus, or combination complexes containing both transferrin and adenovirus, were evaluated. The highest in vitr...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.1993.4.1-17
更新日期:1993-02-01 00:00:00
abstract::Rat myoblast primary cultures were tested as a model for proinsulin synthesis and processing and unregulated insulin delivery for insulin-dependent diabetes mellitus (IDDM) gene therapy. Three human proinsulin cDNA constructs containing genetically engineered furin endoprotease cleavage sites between the B-chain and C...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.1996.7.1-71
更新日期:1996-01-01 00:00:00
abstract::Stereotactic inoculation of a herpes simplex virus (HSV) gene transfer vector into the hippocampus and caudate of rat brain resulted in limited and transient viral replication and the establishment of latency. Virus attenuation was achieved by insertional inactivation of a viral gene, Us3. Insertion of a lacZ reporter...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.1992.3.1-11
更新日期:1992-02-01 00:00:00
abstract::Less than 20% of the protein coding genome is thought to be targetable using small molecules. mRNA therapies are not limited in the same way since in theory, they can silence or edit any gene by encoding CRISPR nucleases, or alternatively, produce any missing protein. Yet not all mRNA therapies are equally likely to s...
journal_title:Human gene therapy
pub_type: 杂志文章
doi:10.1089/hum.2020.137
更新日期:2020-09-01 00:00:00