Genome editing of donor-derived T-cells to generate allogenic chimeric antigen receptor-modified T cells: Optimizing αβ T cell-depleted haploidentical hematopoietic stem cell transplantation

Simple item page
dc.citation.issueNumber3
dc.citation.journalTitleHaematologica
dc.citation.volumeNumber106
dc.contributor.authorWiebking, Volker
dc.contributor.authorLee, Ciaran M.
dc.contributor.authorMostrel, Nathalie
dc.contributor.authorLahiri, Premanjali
dc.contributor.authorBak, Rasmus
dc.contributor.authorBao, Gang
dc.contributor.authorRoncarolo, Maria Grazia
dc.contributor.authorBertaina, Alice
dc.contributor.authorPorteus, Matthew H.
dc.date.accessioned2021-04-21T15:46:12Z
dc.date.available2021-04-21T15:46:12Z
dc.date.issued2021
dc.description.abstractAllogeneic hematopoietic stem cell transplantation is an effective therapy for high-risk leukemias. In children, graft manipulation based on the selective removal of αβ T cells and B cells has been shown to reduce the risk of acute and chronic graft-versus-host disease, thus allowing the use of haploidentical donors which expands the population that allogeneic hematopoietic stem cell transplantation can be used in. Leukemic relapse, however, remains a problem. T cells expressing chimeric antigen receptors can potently eliminate leukemia, including in the central nervous system. We hypothesized that by modifying donor αβ T cells to simultaneously express a CD19-specific chimeric antigen receptors and inactivating the T cell receptor by genome editing, we could create a therapy that enhances the anti-leukemic efficacy of the stem cell transplant without increasing the risk of graft-versus-host disease. Using genome editing with Cas9 ribonucleoprotein and adeno-associated virus serotype 6, we integrate a CD19-specific chimeric antigen receptor in-frame into the TRAC locus. Greater than 90% of cells lost TCR expression, while >75% expressed the CAR. The product was further purified to ultimately have less than 0.05% residual TCR+ cells. In vitro, the CAR T cells efficiently eliminated target cells and produced high cytokine levels when challenged with CD19+ leukemia cells. In vivo, the gene modified T cells eliminated leukemia without causing xenogeneic graft-versus-host disease in a xenograft model. Gene editing was highly specific with no evidence of off-target effects. These data support the concept that the addition of αβ T cell-derived, genome edited T cells expressing CD19-specific chimeric antigen receptors could enhance the anti-leukemic efficacy of αβ T cell-depleted haploidentical hematopoietic stem cell transplantation without increasing the risk of graft-versus-host disease.
dc.identifier.citationWiebking, Volker, Lee, Ciaran M., Mostrel, Nathalie, et al.. "Genome editing of donor-derived T-cells to generate allogenic chimeric antigen receptor-modified T cells: Optimizing αβ T cell-depleted haploidentical hematopoietic stem cell transplantation." <i>Haematologica,</i> 106, no. 3 (2021) Ferrata-Storti Foundation: https://doi.org/10.3324/haematol.2019.233882.
dc.identifier.digital9701-71341-2-10-20210222
dc.identifier.doihttps://doi.org/10.3324/haematol.2019.233882
dc.identifier.urihttps://hdl.handle.net/1911/110289
dc.language.isoeng
dc.publisherFerrata-Storti Foundation
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/
dc.titleGenome editing of donor-derived T-cells to generate allogenic chimeric antigen receptor-modified T cells: Optimizing αβ T cell-depleted haploidentical hematopoietic stem cell transplantation
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
9701-71341-2-10-20210222.pdf
Size:
2.58 MB
Format:
Adobe Portable Document Format
Download
Collections
Faculty Publications
Bioengineering Publications