A5043012920- CAR-T cell therapy research
- Immune Cell Function and Interaction
- CRISPR and Genetic Engineering
- RNA Interference and Gene Delivery
- Viral Infectious Diseases and Gene Expression in Insects
- Acute Myeloid Leukemia Research
- Immune cells in cancer
- Hematopoietic Stem Cell Transplantation
- T-cell and B-cell Immunology
The Ohio State University
2020-2022
Nationwide Children's Hospital
2018-2022
Human primary natural killer (NK) cells are being widely advanced for cancer immunotherapy. However, methods gene editing of these have suffered low transduction rates, high cell death, and loss transgene expression after expansion. Here, we developed a highly efficient method site-specific insertion in NK using CRISPR (Cas9/RNP) AAVs. We compared AAV vectors designed to mediate by different DNA repair mechanisms, homology arm lengths, virus concentrations. then validated the site-directed...
CRISPR/Cas9 technology is accelerating genome engineering in many cell types, but so far, gene delivery and stable modification have been challenging primary NK cells. For example, transgene using lentiviral or retroviral transduction resulted a limited yield of genetically-engineered cells due to substantial procedure-associated apoptosis. We describe here DNA-free method for editing human expanded Cas9 ribonucleoprotein complexes (Cas9/RNPs). This allowed efficient knockout the TGFBR2...
The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that regulates cellular processes in cancer and immunity, including innate immune cell development effector function. However, the transcriptional repertoire through which AHR mediates these effects remains largely unexplored. To elucidate elements directly regulated by natural killer (NK) cells, we performed RNA chromatin immunoprecipitation sequencing on NK cells exposed to agonist or antagonist. We show mature...
CRISPR/Cas9 technology is accelerating genome engineering in many cell types, but so far, gene delivery and stable modification have been challenging primary NK cells. For example, transgene using lentiviral or retroviral transduction resulted a limited yield of genetically-engineered cells due to substantial procedure-associated apoptosis. We describe here DNA-free method for editing human expanded Cas9 ribonucleoprotein complexes (Cas9/RNPs). This allowed efficient knockout the TGFBR2...
Abstract Human peripheral blood natural killer (NK) cells have intense antitumor activity and been used successfully in several clinical trials. Modifying NK with a chimeric antigen receptor (CAR) can improve their targeting increase specificity. Recently, we described an efficient method for gene using Cas9/ribonucleoprotein (RNP) complexes. Here combined this approach single-stranded (ss) or self-complementary (sc) Adeno-associated virus (AAV)-mediated delivery insertion into safe-harbor...
Human peripheral blood natural killer (NK) cells have intense antitumor activity and been used successfully in several clinical trials. Modifying NK with a chimeric antigen receptor (CAR) can improve their targeting increase specificity, but genetic modification of primary human has problematic compared to the ease which T are transduced. Despite some progress made using lentiviral retroviral vectors, efficiency viral transduction remains relatively low. Additionally, these vectors results...
Abstract Human peripheral blood natural killer (NK) cells have strong antitumor activity and been used successfully in several clinical trials. Modifying NK with a chimeric antigen receptor (CAR) can improve their targeting increase specificity. However, genetic modification of has challenging due to the high expression innate sensing mechanisms for viral nucleic acids. Recently, we described an efficient vector-free method using Cas9/ribonucleoprotein complexes gene deletion cells. Here,...
Natural killer (NK) cell determinants predict relapse-free survival after allogeneic hematopoietic transplantation (HCT) for acute myelogenous leukemia, and previous studies have shown a beneficial graft-versus-leukemia effect in patients with juvenile myelomonocytic leukemia (JMML). However, whether NK protection against relapse JMML undergoing HCT is unknown. Therefore, we investigated cell-related donor recipient immunogenetics as of outcomes JMML. Patients (age 0 to <19 years) who...