CRISPR-Cas9 disruption of PD-1 enhances activity of universal EGFRvIII CAR T cells in a preclinical model of human glioblastoma
Authors: Henia Darr
Jason G Sagert
Rebecca C Larson
Genevieve M Gerhard
Irene Scarfò
Andrea Schmidts
Mark B Leick
Marcela V Maus
William T Curry
Stefanie R Bailey
Ana P Castano
Amanda A Bouffard
Bob S Carter
Xiaoling Yu
Daniel B Henderson
Matthew J Frigault
Bryan D Choi
Journal: Journal for immunotherapy of cancer (2019)
Discipline: IMMUNOLOGY
Keywords: CRISPR-Cas systems; EGFRvIII; Glioblastoma; Receptors, chimeric antigen.
Affiliations: Cellular Immunotherapy Program, Cancer Center, Massachusetts General Hospital and Harvard Medical School, 149 13th Street, Room 3.216, Charlestown, Boston, Massachusetts, 02129, USA., Department of Neurosurgery, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA., CRISPR Therapeutics, Cambridge, Massachusetts, USA., Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA., Cellular Immunotherapy Program, Cancer Center, Massachusetts General Hospital and Harvard Medical School, 149 13th Street, Room 3.216, Charlestown, Boston, Massachusetts, 02129, USA. ., Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA. .

Abstract:  Despite remarkable success in the treatment of hematological malignancies, CAR T-cell therapies for solid tumors have floundered, in large part due to local immune suppression and the effects of prolonged stimulation leading to T-cell dysfunction and exhaustion. One mechanism by which gliomas and other cancers can hamper CAR T cells is through surface expression of inhibitory ligands such as programmed cell death ligand 1 (PD-L1). Using the CRIPSR-Cas9 system, we created universal CAR T cells resistant to PD-1 inhibition through multiplexed gene disruption of endogenous T-cell receptor (TRAC), beta-2 microglobulin (B2M) and PD-1 (PDCD1). Triple gene-edited CAR T cells demonstrated enhanced activity in preclinical glioma models. Prolonged survival in mice bearing intracranial tumors was achieved after intracerebral, but not intravenous administration. CRISPR-Cas9 gene-editing not only provides a potential source of allogeneic, universal donor cells, but also enables simultaneous disruption of checkpoint signaling that otherwise impedes maximal antitumor functionality. 

Retracted: No

PubPeer comments:
 Once again this paper fails to[ cite the Jun 2018 paper,] which reports 10 21 death in the Chinese PD-1 CRISPR knockout trial. Not that citing always helps if [you ignore the main issue.] [20 21 people have died ] in that trial. Zero coverage. Like for Mr Saral, who died of condition [he did not have] . Here, 3 gRNAs are used (the Chinese trial just knocked out PD-1) - so more [off-targets] , more dangerous. , 
 And there is no info at all about the gRNAs (no off-target analysis too) - at least the pre-clinical for the UPenn provided that, though they did nothing about the off-targets for PD1 and B2M, not even giving the numbers,[ I did (its easy)] . I know this is a big Univ, and its funded by agreement with CRISPR Therapeutics, but is it allowed to publish papers without giving Methods, esp important pre-clinical ones? , 
 And China has withdrawn [3 PD-1 CRISPR k o trials] - they realize the dangers of [CRISPR, a chromosome shredder] . As I mentioned above, this is worse with 3 edits. ,