Highly Efficient and Selective CAR-Gene Transfer Using CD4- and CD8-Targeted Lentiviral Vectors

Jamali, Arezoo; Kapitza, Laura; Schaser, Thomas; Johnston, Ian; Buchholz, Christian J.; Hartmann, Jessica

doi:10.1016/j.omtm.2019.03.003

Cited by 44 publications

(67 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Studies have shown enhanced TE of Vectofusin-1 compared to other delivery agents such as Tat, Pen, LAH4 derivatives, KH27K, R8, FUSO, polybrene, and retronectin. 328,[351][352][353][354][355] Interestingly, a variation in the histidine sequence order or peptide length resulted in a significant change of bioactivity. A minimum length of 21 amino acids of the LAH4 peptide family was found to be necessary for successful vector delivery.…”

Section: Reviewmentioning

confidence: 99%

Materials promoting viral gene delivery

Kaygisiz

Synatschke

2020

Biomater. Sci.

View full text Add to dashboard Cite

show abstract

Section: Reviewmentioning

confidence: 99%

Materials promoting viral gene delivery

Kaygisiz

Synatschke

2020

Biomater. Sci.

View full text Add to dashboard Cite

show abstract

“…The choice of transduction enhancer and viral envelope protein can tremendously influence transduction efficiencies ( 26 ). Direct comparison of Retronectin, Vectofusin-1, and polybrene revealed that Vectofusin-1 and polybrene are beneficial for the transduction of primary NK cells using VSV-G peusdotyped LV particles and clearly outperform Retronectin.…”

Section: Discussionmentioning

confidence: 99%

“…The transduction enhancer polybrene (Sigma-Aldrich, Germany) was utilized at a concentration of 8 μg/ml and added directly to the cells prior to transduction. Vectofusin-1 (Miltenyi Biotec, Germany) was used according to the manufacturer’s instructions and, as described before ( 26 ), at a final concentration of 10 μg/ml. In brief, Vectofusin-1 and vector particles were diluted in X-Vivo10 medium without additives in a total volume of 50 μl before both solutions were mixed, incubated for 5–10 min at room temperature, and finally added to the NK cells.…”

Section: Methodsmentioning

confidence: 99%

Highly Efficient Generation of Transgenically Augmented CAR NK Cells Overexpressing CXCR4

et al. 2020

Self Cite

View full text Add to dashboard Cite

Natural killer (NK) cells are a noteworthy lymphocyte subset in cancer adoptive cell therapy. NK cells initiate innate immune responses against infections and malignancies with natural cytotoxicity, which is independent of foreign antigen recognition. Based on these substantive features, genetically modifying NK cells is among the prime goals in immunotherapy but is currently difficult to achieve. Recently, we reported a fully human CAR19 construct (huCAR19) with remarkable function in gene-modified T-cells. Here, we show efficient and stable gene delivery of huCAR19 to primary human NK cells using lentiviral vectors with transduction efficiencies comparable to those achieved with NK cell lines. These huCAR19 NK cells display specific and potent cytotoxic activity against target cells. To improve homing of NK cells to the bone marrow, we augmented huCAR19 NK cells with the human CXCR4 gene, resulting in transgenically augmented CAR NK cells (TRACKs). Compared to conventional CAR NK cells, TRACKs exhibit enhanced migration capacity in response to recombinant SDF-1 or bone marrow stromal cells while retaining functional and cytolytic activity against target cells. Based on these promising findings, TRACKs may become a novel candidate for immunotherapeutic strategies in clinical applications.

show abstract

“…Notably, in both lentivirus studies, efficient transduction required activation of the PBMCs. Jamali et al used a transduction-enhancing cationic peptide to further increase the efficiency of the targeted lentivirus system in cultured cells 36 .…”

Section: Discussionmentioning

confidence: 99%

In vivo engineering of lymphocytes after systemic exosome-associated AAV delivery

Breuer

Hanlon

Natasan

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Ex-vivo gene therapy using stem cells or t cells transduced by retroviral or lentiviral vectors has shown remarkable efficacy in the treatment of immunodeficiencies and cancer. However, the process is expensive, technically challenging, and not readily scalable to large patient populations, particularly in underdeveloped parts of the world. Direct in vivo gene therapy would avoid these issues, and such approaches with adeno-associated virus (AAV) vectors have been shown to be safe and efficacious in clinical trials for diseases affecting differentiated tissues such as the liver and CNS. However, the ability to transduce lymphocytes with AAV in vivo after systemic delivery has not been carefully explored. Here, we show that both standard and exosome-associated preparations of AAV8 vectors can effectively transduce a variety of immune cell populations including CD4 + T cells, CD8 + t cells, B cells, macrophages, and dendritic cells after systemic delivery in mice. We provide direct evidence of t cell transduction through the detection of AAV genomes and transgene mRnA, and show that intracellular and transmembrane proteins can be expressed. These findings establish the feasibility of AAV-mediated in vivo gene delivery to immune cells which will facilitate both basic and applied research towards the goal of direct in vivo gene immunotherapies. Gene therapy using ex-vivo transduction of patient-derived cells has revolutionized medicine, particularly in the case of readily accessible blood hematopoietic stem cells or lymphocytes to treat immunodeficiencies or cancer 1-3. However, the laborious workflow and infrastructure required makes these therapies expensive, time-consuming, and unscalable. Moreover, the manipulation of cells outside the body may introduce undesirable phenotypic changes 4. Administering vectors directly into patients would solve most of these problems, potentially expanding the reach of gene therapies to large patient populations anywhere in the world, but unfortunately, such therapies have not advanced due to the absence of a proven and effective delivery system. AAV vectors were thoroughly developed over the past 35 years and are now demonstrating remarkable, life-changing efficacy in clinical trials for several diseases, including recently FDA approved treatments for a form of hereditary blindness and spinal muscular atrophy 5-9. The majority of AAV-mediated gene therapies have focused on differentiated cells such as muscle 10,11 , neurons, astrocytes 12 , and liver 13. AAV has generally been considered inefficient at transducing T cells; however, a recent study revealed that CD3 + T cells are a natural reservoir for multiple wild type AAV serotypes 14. Here we demonstrate that AAV8 vectors can mediate transgene expression in multiple immune cell types after systemic delivery in mice, providing an important proof of concept for developing these vectors for use in in vivo therapies.

show abstract

Highly Efficient and Selective CAR-Gene Transfer Using CD4- and CD8-Targeted Lentiviral Vectors

Cited by 44 publications

References 32 publications

Materials promoting viral gene delivery

Materials promoting viral gene delivery

Highly Efficient Generation of Transgenically Augmented CAR NK Cells Overexpressing CXCR4

In vivo engineering of lymphocytes after systemic exosome-associated AAV delivery

Contact Info

Product

Resources

About