Inosine induces stemness features in CAR-T cells and enhances potency

Dorota D. Klysz; Carley Fowler; Meena Malipatlolla; Lucille Stuani; Katherine A. Freitas; Yiyun Chen; Stefanie Meier; Bence Daniel; Katalin Sandor; Peng Xu; Jing Huang; Louai Labanieh; Vimal Keerthi; Amaury Leruste; Malek Bashti; Janette Mata-Alcazar; Nikolaos Gkitsas; Justin A. Guerrero; Chris Fisher; Sunny Patel; Kyle Asano; Shabnum Patel; Kara L. Davis; Ansuman T. Satpathy; Steven A. Feldman; Elena Sotillo; Crystal L. Mackall

doi:10.1016/j.ccell.2024.01.002

Inosine induces stemness features in CAR-T cells and enhances potency

Dorota D. Klysz
, Carley Fowler
, Meena Malipatlolla
, Lucille Stuani
, Katherine A. Freitas
, Yiyun Chen
, Stefanie Meier
, Bence Daniel
, Katalin Sandor
, Peng Xu
, Jing Huang
, Louai Labanieh
, Vimal Keerthi
, Amaury Leruste
, Malek Bashti
, Janette Mata-Alcazar
, Nikolaos Gkitsas
, Justin A. Guerrero
, Chris Fisher
, Sunny Patel

Kyle Asano, Shabnum Patel, Kara L. Davis, Ansuman T. Satpathy, Steven A. Feldman, Elena Sotillo, Crystal L. Mackall

Research output: Contribution to journal › Article › peer-review

132 Scopus citations

Abstract

Adenosine (Ado) mediates immune suppression in the tumor microenvironment and exhausted CD8⁺ CAR-T cells express CD39 and CD73, which mediate proximal steps in Ado generation. Here, we sought to enhance CAR-T cell potency by knocking out CD39, CD73, or adenosine receptor 2a (A2aR) but observed only modest effects. In contrast, overexpression of Ado deaminase (ADA-OE), which metabolizes Ado to inosine (INO), induced stemness and enhanced CAR-T functionality. Similarly, CAR-T cell exposure to INO augmented function and induced features of stemness. INO induced profound metabolic reprogramming, diminishing glycolysis, increasing mitochondrial and glycolytic capacity, glutaminolysis and polyamine synthesis, and reprogrammed the epigenome toward greater stemness. Clinical scale manufacturing using INO generated enhanced potency CAR-T cell products meeting criteria for clinical dosing. These results identify INO as a potent modulator of CAR-T cell metabolism and epigenetic stemness programming and deliver an enhanced potency platform for cell manufacturing.

Original language	English
Pages (from-to)	266-282.e8
Journal	Cancer Cell
Volume	42
Issue number	2
DOIs	https://doi.org/10.1016/j.ccell.2024.01.002
State	Published - 12 Feb 2024
Externally published	Yes

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Klysz, D. D., Fowler, C., Malipatlolla, M., Stuani, L., Freitas, K. A., Chen, Y., Meier, S., Daniel, B., Sandor, K., Xu, P., Huang, J., Labanieh, L., Keerthi, V., Leruste, A., Bashti, M., Mata-Alcazar, J., Gkitsas, N., Guerrero, J. A., Fisher, C., ... Mackall, C. L. (2024). Inosine induces stemness features in CAR-T cells and enhances potency. Cancer Cell, 42(2), 266-282.e8. https://doi.org/10.1016/j.ccell.2024.01.002

@article{6cbb4d1c1bce4dcea966c30e80279f1f,

title = "Inosine induces stemness features in CAR-T cells and enhances potency",

abstract = "Adenosine (Ado) mediates immune suppression in the tumor microenvironment and exhausted CD8+ CAR-T cells express CD39 and CD73, which mediate proximal steps in Ado generation. Here, we sought to enhance CAR-T cell potency by knocking out CD39, CD73, or adenosine receptor 2a (A2aR) but observed only modest effects. In contrast, overexpression of Ado deaminase (ADA-OE), which metabolizes Ado to inosine (INO), induced stemness and enhanced CAR-T functionality. Similarly, CAR-T cell exposure to INO augmented function and induced features of stemness. INO induced profound metabolic reprogramming, diminishing glycolysis, increasing mitochondrial and glycolytic capacity, glutaminolysis and polyamine synthesis, and reprogrammed the epigenome toward greater stemness. Clinical scale manufacturing using INO generated enhanced potency CAR-T cell products meeting criteria for clinical dosing. These results identify INO as a potent modulator of CAR-T cell metabolism and epigenetic stemness programming and deliver an enhanced potency platform for cell manufacturing.",

author = "Klysz, \{Dorota D.\} and Carley Fowler and Meena Malipatlolla and Lucille Stuani and Freitas, \{Katherine A.\} and Yiyun Chen and Stefanie Meier and Bence Daniel and Katalin Sandor and Peng Xu and Jing Huang and Louai Labanieh and Vimal Keerthi and Amaury Leruste and Malek Bashti and Janette Mata-Alcazar and Nikolaos Gkitsas and Guerrero, \{Justin A.\} and Chris Fisher and Sunny Patel and Kyle Asano and Shabnum Patel and Davis, \{Kara L.\} and Satpathy, \{Ansuman T.\} and Feldman, \{Steven A.\} and Elena Sotillo and Mackall, \{Crystal L.\}",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors",

year = "2024",

month = feb,

day = "12",

doi = "10.1016/j.ccell.2024.01.002",

language = "English",

volume = "42",

pages = "266--282.e8",

journal = "Cancer Cell",

issn = "1535-6108",

publisher = "Cell Press",

number = "2",

}

Klysz, DD, Fowler, C, Malipatlolla, M, Stuani, L, Freitas, KA, Chen, Y, Meier, S, Daniel, B, Sandor, K, Xu, P, Huang, J, Labanieh, L, Keerthi, V, Leruste, A, Bashti, M, Mata-Alcazar, J, Gkitsas, N, Guerrero, JA, Fisher, C, Patel, S, Asano, K, Patel, S, Davis, KL, Satpathy, AT, Feldman, SA, Sotillo, E & Mackall, CL 2024, 'Inosine induces stemness features in CAR-T cells and enhances potency', Cancer Cell, vol. 42, no. 2, pp. 266-282.e8. https://doi.org/10.1016/j.ccell.2024.01.002

TY - JOUR

T1 - Inosine induces stemness features in CAR-T cells and enhances potency

AU - Klysz, Dorota D.

AU - Fowler, Carley

AU - Malipatlolla, Meena

AU - Stuani, Lucille

AU - Freitas, Katherine A.

AU - Chen, Yiyun

AU - Meier, Stefanie

AU - Daniel, Bence

AU - Sandor, Katalin

AU - Xu, Peng

AU - Huang, Jing

AU - Labanieh, Louai

AU - Keerthi, Vimal

AU - Leruste, Amaury

AU - Bashti, Malek

AU - Mata-Alcazar, Janette

AU - Gkitsas, Nikolaos

AU - Guerrero, Justin A.

AU - Fisher, Chris

AU - Patel, Sunny

AU - Asano, Kyle

AU - Patel, Shabnum

AU - Davis, Kara L.

AU - Satpathy, Ansuman T.

AU - Feldman, Steven A.

AU - Sotillo, Elena

AU - Mackall, Crystal L.

PY - 2024/2/12

Y1 - 2024/2/12

N2 - Adenosine (Ado) mediates immune suppression in the tumor microenvironment and exhausted CD8+ CAR-T cells express CD39 and CD73, which mediate proximal steps in Ado generation. Here, we sought to enhance CAR-T cell potency by knocking out CD39, CD73, or adenosine receptor 2a (A2aR) but observed only modest effects. In contrast, overexpression of Ado deaminase (ADA-OE), which metabolizes Ado to inosine (INO), induced stemness and enhanced CAR-T functionality. Similarly, CAR-T cell exposure to INO augmented function and induced features of stemness. INO induced profound metabolic reprogramming, diminishing glycolysis, increasing mitochondrial and glycolytic capacity, glutaminolysis and polyamine synthesis, and reprogrammed the epigenome toward greater stemness. Clinical scale manufacturing using INO generated enhanced potency CAR-T cell products meeting criteria for clinical dosing. These results identify INO as a potent modulator of CAR-T cell metabolism and epigenetic stemness programming and deliver an enhanced potency platform for cell manufacturing.

AB - Adenosine (Ado) mediates immune suppression in the tumor microenvironment and exhausted CD8+ CAR-T cells express CD39 and CD73, which mediate proximal steps in Ado generation. Here, we sought to enhance CAR-T cell potency by knocking out CD39, CD73, or adenosine receptor 2a (A2aR) but observed only modest effects. In contrast, overexpression of Ado deaminase (ADA-OE), which metabolizes Ado to inosine (INO), induced stemness and enhanced CAR-T functionality. Similarly, CAR-T cell exposure to INO augmented function and induced features of stemness. INO induced profound metabolic reprogramming, diminishing glycolysis, increasing mitochondrial and glycolytic capacity, glutaminolysis and polyamine synthesis, and reprogrammed the epigenome toward greater stemness. Clinical scale manufacturing using INO generated enhanced potency CAR-T cell products meeting criteria for clinical dosing. These results identify INO as a potent modulator of CAR-T cell metabolism and epigenetic stemness programming and deliver an enhanced potency platform for cell manufacturing.

UR - https://www.scopus.com/pages/publications/85184025054

U2 - 10.1016/j.ccell.2024.01.002

DO - 10.1016/j.ccell.2024.01.002

M3 - Article

C2 - 38278150

AN - SCOPUS:85184025054

SN - 1535-6108

VL - 42

SP - 266-282.e8

JO - Cancer Cell

JF - Cancer Cell

IS - 2

ER -

Inosine induces stemness features in CAR-T cells and enhances potency

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