TY - JOUR
T1 - Large-scale GMP-compliant CRISPR-Cas9-mediated deletion of the glucocorticoid receptor in multivirus-specific T cells
AU - Basar, Rafet
AU - Daher, May
AU - Uprety, Nadima
AU - Gokdemir, Elif
AU - Alsuliman, Abdullah
AU - Ensley, Emily
AU - Ozcan, Gonca
AU - Mendt Vilchez, Mayela Carolina
AU - Sanabria, Mayra Hernandez
AU - Kerbauy, Lucila Nassif
AU - Cortes, Ana Karen Nunez
AU - Li, Li
AU - Banerjee, Pinaki P.
AU - Muniz-Feliciano, Luis
AU - Acharya, Sunil
AU - Fowlkes, Natalie W.
AU - Lu, Junjun
AU - Li, Sufang
AU - Mielke, Stephan
AU - Kaplan, Mecit
AU - Nandivada, Vandana
AU - Bdaiwi, Mustafa
AU - Kontoyiannis, Alexander D.
AU - Li, Ye
AU - Liu, Enli
AU - Ang, Sonny
AU - Marin, David
AU - Brunetti, Lorenzo
AU - Gundry, Michael C.
AU - Turk, Rolf
AU - Schubert, Mollie S.
AU - Rettig, Garrett R.
AU - McNeill, Matthew S.
AU - Kurgan, Gavin
AU - Behlke, Mark A.
AU - Champlin, Richard
AU - Shpall, Elizabeth J.
AU - Rezvani, Katayoun
N1 - Funding Information:
Acknowledgments This work was supported by the National Institutes of Health, National Cancer Institute (CA061508-21A1 and CA211044-01) and the MD Anderson Cancer Center Acute Myeloid Leukemia (AML) Moon Shot Program.
Publisher Copyright:
© 2020 by The American Society of Hematology.
PY - 2020/7/28
Y1 - 2020/7/28
N2 - Virus-specific T cells have proven highly effective for the treatment of severe and drugrefractory infections after hematopoietic stem cell transplant (HSCT). However, the efficacy of these cells is hindered by the use of glucocorticoids, often given to patients for the management of complications such as graft-versus-host disease. To address this limitation, we have developed a novel strategy for the rapid generation of good manufacturing practice (GMP)-grade glucocorticoid-resistant multivirus-specific T cells (VSTs) using clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) gene-editing technology. We have shown that deleting the nuclear receptor subfamily 3 group C member 1 (NR3C1; the gene encoding for the glucocorticoid receptor) renders VSTs resistant to the lymphocytotoxic effect of glucocorticoids. NR3C1-knockout (KO) VSTs kill their targets and proliferate successfully in the presence of high doses of dexamethasone both in vitro and in vivo. Moreover, we developed a protocol for the rapid generation of GMP-grade NR3C1 KO VSTs with high on-target activity and minimal off-target editing. These genetically engineered VSTs promise to be a novel approach for the treatment of patients with life-threatening viral infections post-HSCT on glucocorticoid therapy.
AB - Virus-specific T cells have proven highly effective for the treatment of severe and drugrefractory infections after hematopoietic stem cell transplant (HSCT). However, the efficacy of these cells is hindered by the use of glucocorticoids, often given to patients for the management of complications such as graft-versus-host disease. To address this limitation, we have developed a novel strategy for the rapid generation of good manufacturing practice (GMP)-grade glucocorticoid-resistant multivirus-specific T cells (VSTs) using clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) gene-editing technology. We have shown that deleting the nuclear receptor subfamily 3 group C member 1 (NR3C1; the gene encoding for the glucocorticoid receptor) renders VSTs resistant to the lymphocytotoxic effect of glucocorticoids. NR3C1-knockout (KO) VSTs kill their targets and proliferate successfully in the presence of high doses of dexamethasone both in vitro and in vivo. Moreover, we developed a protocol for the rapid generation of GMP-grade NR3C1 KO VSTs with high on-target activity and minimal off-target editing. These genetically engineered VSTs promise to be a novel approach for the treatment of patients with life-threatening viral infections post-HSCT on glucocorticoid therapy.
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U2 - 10.1182/bloodadvances.2020001977
DO - 10.1182/bloodadvances.2020001977
M3 - Article
C2 - 32717029
AN - SCOPUS:85090589429
SN - 2473-9529
VL - 4
SP - 3357
EP - 3367
JO - Blood Advances
JF - Blood Advances
IS - 14
ER -