Engineered endothelial cells expressing an ETS transcription factor

Inventors

Nolan, Daniel Joseph • Finnegan, Paul William • Ginsberg, Michael Daniel • Davis, Claude Geoffrey

Assignees

Angiocrine Bioscience Inc

Abstract

In some aspects the present invention relates to engineered endothelial cells, such as E40RF1+ ETV2+ engineered endothelial cells. In other aspects the present invention relates to methods of making such engineered endothelial cells, and methods of using such engineered endothelial cells, for example in co-culture applications.

Core Innovation

The invention relates to engineered human umbilical vein endothelial cells (HUVECs) that co-express adenovirus E4 region E4ORF1 and an ETS family transcription factor ETV2, generating E4ORF1+ETV2+ engineered endothelial cells. The document further describes compositions in which these engineered endothelial cells are used together with hematopoietic stem or progenitor cells (HSPCs), including isolated compositions comprising E4ORF1+ETV2+ HUVECs and HSPCs. A key restriction described is that the engineered HUVECs do not comprise an entire adenovirus E4 region.

The document states that co-expression of E4ORF1 and ETV2 unexpectedly improves support for hematopoietic stem/progenitor cell expansion compared with alternative endothelial co-expression conditions described in the partial content, including ETV2-only or E4ORF1-only contexts. The engineered endothelial approach is described as providing enhanced expansion outcomes for HSPCs and preferential expansion of more primitive HSPC subsets.

Embodiments described in the partial content include isolated or substantially pure engineered endothelial cell populations, and therapeutic compositions that optionally comprise a carrier solution and HSPCs. The partial content also describes culture and co-culture approaches, including feeder cell layer use and conditioned-medium approaches, to maintain and expand HSPCs with the E4ORF1+ETV2+ engineered endothelial cells.

The partial content further describes that co-culture with E4ORF1+ETV2+ endothelial cells yields greater CD34+ HSPC expansion and more primitive phenotypes, including CD34+high and CD45RA−, compared with the comparator endothelial conditions referenced in the partial content. The document links these observed expansion and phenotype outcomes to the co-expression of E4ORF1 and ETV2 in engineered endothelial cells.

Claims Coverage

The partial content provides two independent claims, covering an isolated composition containing E4ORF1+ETV2+ engineered HUVECs and HSPCs with a restriction on adenovirus E4 region content, and a culture method for maintaining or expanding HSPCs by culturing them in the same vessel as E4ORF1+ETV2+ engineered HUVECs. Across the independent claims, the core inventive theme is the use of E4ORF1+ETV2+ engineered HUVECs in combination with HSPCs, including a defined constraint on which adenovirus E4 elements are present in the engineered HUVECs.

The claim coverage centers on using E4ORF1+ETV2+ engineered HUVECs with HSPCs, including an explicit restriction that the HUVECs do not comprise an entire adenovirus E4 region, and a culture approach where engineered E4ORF1+ETV2+ HUVECs are cultured in the same vessel as the HSPCs.

Stated Advantages

Documented Applications