Gene delivery in a mouse xenograft of a retargeted retrovirus to a solid 143B osteosarcoma

Zhang, Xia; Sarangi, Anindita; Wu, Daitze; Kanduri, Jaya; Roth, Monica J.

doi:10.1186/1743-422x-10-194

Cited by 2 publications

(4 citation statements)

References 21 publications

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“…A variety of approaches have shown that retargeting of retroviral particle entry can be achievedvia alteration of the viral–host interactions, including single-chain antibodies [7]. Our laboratory has developed a screen to identify retroviral Env capable of productively infecting target cells through randomizing an 11 amino acid region of the receptor binding domain of feline leukemia virus (FeLV) [41–45] and established the effectiveness of intratumor delivery [46]. Additionally, successful retargeting for Sindbis Env pseudotyped onto lentiviral particles has also been achieved with the insertion of the Protein A ZZ domain and sandwiching targeting antibodies to the modified particles.…”

Section: Discussionmentioning

confidence: 99%

MLV based viral-like-particles for delivery of toxic proteins and nuclear transcription factors

Wu¹,

Roth²

2014

Biomaterials

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We have developed nanoparticles based on Murine Leukemia Virus virus-like-particles (VLPs) that efficiently deliver therapeutic bioactive proteins in their native state into target cells. Nuclear transcription factors and toxic proteins were incorporated into the VLPs from stable producer cells without transducing viral-encoded genetic material. Delivery of nuclear transcription factors required incorporation of nuclear export signals (NESs) into the vector backbone for the efficient formation of VLPs. In the presence of an appropriate targeting Env glycoprotein, transcription factors delivered and activated nuclear transcription in the target cells. Additionally, we show delivery of the bacterial toxin, MazF, which is an ACA-specific mRNA interferase resulted in the induction of cell death. The stable producer cells are protected from the toxin through co-expression of the anti-toxin MazE and continuously released MazF incorporating VLPs. This highly adaptable platform can be harnessed to alter and regulate cellular processes by bioactive protein delivery.

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Section: Discussionmentioning

confidence: 99%

MLV based viral-like-particles for delivery of toxic proteins and nuclear transcription factors

Wu¹,

Roth²

2014

Biomaterials

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“…Previous techniques produced a limited number of high-affinity Env isolates in each round of screening (18)(19)(20)(21)(22). Additional studies indicated that the pRVL vector (22) used to build the library of Envs had a high frequency of aberrant splicing into the internal SV40 promoter region, eliminating the packaging of the RNA into viral particles (28).…”

Section: Modification Of the Vector Backbone For Improved Library Scrmentioning

confidence: 99%

“…Finally, the BamHI CP Env cassette was inserted into the BamHI site to generate the pRVL-M1-CP. pRVL-CP-NWPRE was constructed by digesting pRVL-CP with BamHI and inserting the 620-bp woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) BamHI-BglII fragment previously described (28). Env was subsequently reinserted back using the BamHI site.…”

Section: Cell Linesmentioning

confidence: 99%

“…In order to create the library vector SD-env-SA-neo-CWPRE, a point mutation was introduced to destroy a BbsI site in the WPRE sequence, using primers FneoXWPRE/RWPRE560 and FWPRE539/RDtWPREp4. The parental template used, HIV2gtmtWPRE (28), encoded a substitution of the ATG start codon in the WPRE sequence, to abrogate translation of the woodchuck hepatitis virus X protein (29). The BbsI Ϫ WPRE sequence was introduced into the plasmid vector downstream of the neomycin resistance gene by overlapping PCR.…”

Section: Cell Linesmentioning

confidence: 99%

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Role of Cysteines in Stabilizing the Randomized Receptor Binding Domains within Feline Leukemia Virus Envelope Proteins

Valdivieso-Torres

Sarangi

Whidby

et al. 2016

J Virol

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Retargeting of gammaretroviral envelope proteins has shown promising results in the isolation of novel isolates with therapeutic potential. However, the optimal conditions required to obtain high-affinity retargeted envelope proteins with narrow tropism are not understood. This study highlights the advantage of constrained peptides within receptor binding domains and validates the random library screening technique of obtaining novel retargeted Env proteins. Using a modified vector backbone to screen the envelope libraries on 143B osteosarcoma cells, three novel and unique retargeted envelopes were isolated. The use of complex disulfide bonds within variable regions required for receptor binding is found within natural gammaretroviral envelope isolates. Interestingly, two of the isolates, named AII and BV2, have a pair of cysteines located within the randomized region of 11 amino acids similar to that identified within the CP Env, an isolate identified in a previous Env library screen on the human renal carcinoma Caki-1 cell line. The amino acids within the randomized region of AII and BV2 envelopes that are essential for viral infection have been identified in this study and include these cysteine residues. Through mutagenesis studies, the putative disulfide bond pairs including and beyond the randomized region were examined. In parallel, the disulfide bonds of CP Env were identified using mass spectrometry. The results indicate that this pair of cysteines creates the structural context to position key hydrophobic (F and W) and basic (K and H) residues critical for viral titer and suggest that AII, BV2, and CP internal cysteines bond together in distinct ways. T he specificity of retroviral vectors is initially conferred by the envelope (Env) protein present on the surface of virus particles. This Env protein is responsible for binding to a host cell receptor, thereby initiating virus entry. In gammaretroviruses, the Env protein is composed of a transmembrane protein (TM) and a surface protein (SU). For murine leukemia virus (MLV) and feline leukemia virus (FeLV) SUs, primary receptor binding is localized to the N-terminal half of SU (1, 2, 12). Critical residues for specificity are encoded within the N-terminal variable regions, VRA and VRB. Secondary receptor binding sites that promote viral infection have also been identified within the SU C terminus (3-5).FeLV Envs are classified as A, B, and C, originally based on interference assays (6, 7). The receptors for these three major subgroups have been identified as THTR1 (8), PiT1 (9), and FLVCR1 (10, 11), respectively, molecularly confirming the distinct receptor usage for each subgroup. For FeLV-A, 19 residues in the VRA can be replaced with 16 residues of FeLV-C VRA to sufficiently alter the binding specificity, thus switching receptor usage (12). In FeLV-A, the VRA and VRB each include a pair of cysteines, which have the potential to form intramolecular disulfide bonds (5). The cysteine contents of MLV and FeLV-B SUs are more complex; ecotropic MLV ...

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Gene delivery in a mouse xenograft of a retargeted retrovirus to a solid 143B osteosarcoma

Cited by 2 publications

References 21 publications

MLV based viral-like-particles for delivery of toxic proteins and nuclear transcription factors

MLV based viral-like-particles for delivery of toxic proteins and nuclear transcription factors

Role of Cysteines in Stabilizing the Randomized Receptor Binding Domains within Feline Leukemia Virus Envelope Proteins

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