Transiently immortalized cells for use in gene therapy
AssigneeHeart Biosystems GmbH
InventorsE. Edward Baetge, Shou Wong, Philippe Dupraz, Bernard Thorens
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Abstract
The invention provides methods and compositions for expanding cells that are not abundant or are difficult to obtain in pure form in culture, are in short supply (e.g., human cells), or have brief lifetimes in culture, using fusion polypeptide. The fusion polypeptide has a first region having the transport function of herpesviral VP22 protein or human immunodeficiency virus (HIV) TAT protein, and a second region with a polypeptide having cell immortalization activity, a polypeptide having telomerase-specific activity, or a polypeptide having telomerase gene activation activity. The resulting cells of the invention are suitable for use in cell therapy.
Claims10
A fusion polypeptide, comprising:
- (i) a first polypeptide having the transport function of herpesviral VP22 protein or human immunodeficiency virus (HIV) TAT protein, and
- (ii) a second polypeptide selected from
- (a) a polypeptide having cell immortalization activity,
- (b) a polypeptide having telomerase-specific activity, and
- (c) a polypeptide having telomerase gene activation activity.
A method of transiently immortalizing a cell comprising the step of expanding the cell in growth medium containing a fusion protein comprising a first polypeptide having the transport function of herpesviral VP22 protein or human immunodeficiency virus (HIV) TAT protein and a second polypeptide having cell immortalization activity, wherein the fusion protein is taken up by the cell to result in proliferation of the cell, and wherein growing the immortalized cell in growth medium that does not contain the fusion protein terminates the proliferative effects of the fusion protein.
A method of transiently telomerizing a cell comprising the step of exposing the cell to growth medium containing a fusion protein comprising a first polypeptide having the transport function of herpesviral BP22 protein or human immunodeficiency virus (HIV) TAT protein and a second polypeptide having telomerase-specific activity, wherein growing the immortalized cell in growth medium that does not contain the fusion protein terminates the telomerizing effect of the fusion protein.
A method of increasing the replicative capacity of a cell comprising the step of expanding the cell in growth medium containing:
- (i) a first fusion protein comprising a first polypeptide having the transport function of herpesviral VP22 protein or human immunodeficiency virus (HIV) TAT protein and a second polypeptide having cell immortalization activity, and
- (ii) a second fusion protein comprising a first polypeptide having the transport function of herpesviral VP22 protein or human immunodeficiency virus (HIV) TAT protein and a second polypeptide having telomerase-specific activity,
- wherein growing the cell in growth medium that does not contain the first and second fusion proteins terminates the immortalizing and the telomerizing effects of the fusion proteins.
Dependencies are inferred from the claim text.