Retroviral transfer of genes into canine hemopoietic progenitor cells in culture: a model for human gene therapy.

Kwok, William W.; Schuening, FG; Stead, RB; Miller, A D

doi:10.1073/pnas.83.12.4552

Cited by 78 publications

(34 citation statements)

References 23 publications

(28 reference statements)

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“…The ecotropic viral envelope receptor (29) was cloned into pBabe (30) and transfected into PA317 amphotropic packaging cells (31). Retroviral-conditioned medium was used to infect human U251 astrocytoma and A673 PNET cell lines (both negative for Rig protein expression; C.A.E.…”

Section: Generation Of Neural Tumor Cells Harboring An Inducible Rig mentioning

confidence: 99%

Rig is a novel Ras-related protein and potential neural tumor suppressor

Ellis

Vos

Howell

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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The Ras superfamily consists of a large group of monomeric GTPases demonstrating homology to Ras oncoproteins. Although structurally similar, Ras-superfamily proteins are functionally diverse. Whereas some members exhibit oncogenic properties, others may serve as tumor suppressors. We have identified a novel Ras-related protein that suppresses cell growth and have designated it Rig (Ras-related inhibitor of cell growth). Overexpression of Rig inhibited Ras-mediated cellular transformation and activation of downstream signaling in NIH 3T3 cells. rig mRNA is expressed at high levels in normal cardiac and neural tissue. However, Rig protein expression is frequently lost or down-regulated in neural tumor-derived cell lines and primary human neural tumors. Moreover, expression of exogenous Rig in human astrocytoma cells suppressed growth. Rig has a C-terminal CAAX motif that codes for posttranslational modification by both farnesyl and geranylgeranyl isoprenoid lipids. Consequently, Rig may play a role in the cellular response to farnesyl transferase inhibitors. Rig bears 63% overall sequence homology to a recently described Ras-family member Noey2, a tumor suppressor in breast and ovarian tissue. Therefore, Rig and Noey2 may represent a new subfamily of Ras-like tumor suppressors. S mall, Ras-related GTPases form a large superfamily of structurally related proteins in mammalian cells (1). These proteins are typically regulated by guanine nucleotide binding and undergo C-terminal isoprenylation. Ras-superfamily proteins mediate pleiotropic cellular effects ranging from growth control to cytoskeletal rearrangements, various aspects of intracellular transport, cell survival, and apoptosis (2, 3). Although the Ras-, Ral-, Rit-, and Rho-subfamily proteins function as oncoproteins (4, 5), Noey2͞Ahri1 and Rap1A have been described as tumor suppressors (6, 7). Thus, even closely related Ras-superfamily members can exert quite different biological effects.The relationship between structure and function of Rasrelated proteins is now sufficiently well understood that certain biological and biochemical characteristics may be inferred simply from an examination of their primary amino acid sequence. For example, the region of these proteins essential for effector interaction (the effector domain) has been identified and the residues important for particular effector interactions characterized (8). Furthermore, many Ras-related proteins contain a C-terminal CAAX motif (cysteine-aliphatic-aliphatic-X) that serves as a target for posttranslational isoprenoid lipid modification (9). It is now known that the X amino acid residue is critical for determining the type of covalent isoprenylation, farnesyl (F) (15-carbon) or geranylgeranyl (GG) (20-carbon). Typically, serine, phenylalanine, methionine, and cysteine residues are substrates for F transferases whereas GG transferases recognize leucine and phenylalanine residues (9, 10).The vast majority of Ras-related proteins undergo GG isoprenoid lipid modification whereas the relatively ra...

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Section: Generation Of Neural Tumor Cells Harboring An Inducible Rig mentioning

confidence: 99%

Rig is a novel Ras-related protein and potential neural tumor suppressor

Ellis

Vos

Howell

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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“…Gene transfer into primitive stem cells should result in the continued expression of the gene in blood cells, including T lymphocytes. Retrovirus-mediated transfer and expression of genes in hematopoietic progenitor cells of humans (7,8), monkeys (9), dogs (10), and mice (11,12) have been demonstrated in culture. Gene transfer into marrow stem cells of mice followed by continued expression of the transferred gene in progeny hematopoietic cells in vivo has also been reported (11)(12)(13)(14).…”

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confidence: 99%

Design of vectors for efficient expression of human purine nucleoside phosphorylase in skin fibroblasts from enzyme-deficient humans.

Osborne

Miller

1988

Proc. Natl. Acad. Sci. U.S.A.

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Purine nucleoside phosphorylase (PNP; purine-nucleoside orthophosphate ribosyltransferase, EC 2.4.2.1) deficiency is an inherited disorder associated with a severe immune defect that is fatal. Enzyme replacement therapy is an attractive approach to treatment of this disease. To this aim we constructed retroviral vectors containing a human PNP cDNA and a selectable gene encoding neomycin phosphotransferase. PNP expression was controlled by either the early promoter from simian virus 40, the immediate early promoter from human cytomegalovirus, or the retroviral promoter. Cultured-skin fibroblasts from two unrelated PNP. deficient patients that were infected with these vectors expressed mean PNP activities of0.03, 0.74, and 5.9 pmol/hr per mg of protein, respectively. The latter infectants had PNP activities eight times the level of 0.74 ,umol/hr per mg of protein observed in normal skin fibroblasts, enabling rapid metabolism of exogenous deoxyguanosine, the cytotoxic metabolite that accumulates in the plasma of PNP-deficient patients. These experiments indicate that viral long terminal repeat was the strongest promoter for expression of PNP and suggest the potential of human skin fibroblasts as vehicles for therapeutic gene expression.

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“…The vector-infected MPS VII hematopoietic cells apparently expressed normal amounts of GUS enzymatic activity (Table 3). This was an unexpected result because retroviral vector transfer of the neo gene to canine bone marrow cells usually has been found to be inefficient when assayed for G418-resistant colony formation (33)(34)(35). To determine if the vector-encoded enzyme was present in all of the vectortreated bone marrow cells, histochemical staining for GUS enzymatic activity (28,29) was used to visualize individual cells in infected populations (Fig.…”

Section: Resultsmentioning

confidence: 99%