Benign tumors in TSC are amenable to treatment by GD3 CAR T cells in mice

Ancy Thomas; Saurav Sumughan; Emilia R. Dellacecca; Rohan S. Shivde; Nicola Lancki; Zhussipbek Mukhatayev; Cristina C. Vaca; Fei Han; Levi Barse; Steven W. Henning; Jesus Zamora-Pineda; Suhail Akhtar; Nikhilesh Gupta; Jasmine O. Zahid; Stephanie R. Zack; Prathyaya Ramesh; Dinesh Jaishankar; Agnes S.Y. Lo; Joel Moss; Maria M. Picken; Thomas N. Darling; Denise M. Scholtens; Daniel F. Dilling; Richard P. Junghans; I. Caroline Le Poole

doi:10.1172/jci.insight.152014

Benign tumors in TSC are amenable to treatment by GD3 CAR T cells in mice

Ancy Thomas, Saurav Sumughan, Emilia R. Dellacecca, Rohan S. Shivde, Nicola Lancki, Zhussipbek Mukhatayev, Cristina C. Vaca, Fei Han, Levi Barse, Steven W. Henning, Jesus Zamora-Pineda, Suhail Akhtar, Nikhilesh Gupta, Jasmine O. Zahid, Stephanie R. Zack, Prathyaya Ramesh, Dinesh Jaishankar, Agnes S.Y. Lo, Joel Moss, Maria M. PickenThomas N. Darling, Denise M. Scholtens, Daniel F. Dilling, Richard P. Junghans, I. Caroline Le Poole^*

^*Corresponding author for this work

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

7 Scopus citations

Abstract

Mutations underlying disease in tuberous sclerosis complex (TSC) give rise to tumors with biallelic mutations in TSC1 or TSC2 and hyperactive mammalian target of rapamycin complex 1 (mTORC1). Benign tumors might exhibit de novo expression of immunogens, targetable by immunotherapy. As tumors may rely on ganglioside D3 (GD3) expression for mTORC1 activation and growth, we compared GD3 expression in tissues from patients with TSC and controls. GD3 was overexpressed in affected tissues from patients with TSC and also in aging Tsc2+/- mice. As GD3 overexpression was not accompanied by marked natural immune responses to the target molecule, we performed preclinical studies with GD3 chimeric antigen receptor (CAR) T cells. Polyfunctional CAR T cells were cytotoxic toward GD3-overexpressing targets. In mice challenged with Tsc2-/- tumor cells, CAR T cells substantially and durably reduced the tumor burden, correlating with increased T cell infiltration. We also treated aged Tsc2+/- heterozygous (>60 weeks) mice that carry spontaneous Tsc2-/- tumors with GD3 CAR or untransduced T cells and evaluated them at endpoint. Following CAR T cell treatment, the majority of mice were tumor free while all control animals carried tumors. The outcomes demonstrate a strong treatment effect and suggest that targeting GD3 can be successful in TSC.

Original language	English (US)
Article number	e152014
Journal	JCI Insight
Volume	6
Issue number	22
DOIs	https://doi.org/10.1172/jci.insight.152014
State	Published - Nov 22 2021

Funding

The single-cell cytokine analysis was performed at the Immunotherapy Assessment Core/TEST IT, North-western University. Tissue imaging work was performed at the Northwestern University Center for Advanced Microscopy supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. The TEST IT core is supported by NIH P30AR075049 to AS Paller. The flow cytometry was carried out at the Northwestern University\u2013Flow Cytometry Core Facility supported by Cancer Center Support Grant (NCI CA060553). Cytotoxicity assay using IncuCyte S3 was performed in the Analytical bioNano-Technology Equipment Core at Northwestern University. The U.S. Army Research Office, the U.S. Army Medical Research and Materiel Command, and Northwestern University provided funding to develop this facility, and ongoing support is being received from the Soft and Hybrid Nanotechnology Experimental Resource (NSF ECCS-1542205). Histology services were provided by the Northwestern University Mouse Histology and Phenotyping Laboratory, which is supported by NCI P30-CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. These studies were supported by a Department of Defense grant W81XWH-17-TSCRP-CTRA and National Cancer Institute (NCI) RO1 CA191317 to ICLP. The authors acknowledge material support from the NCI Biological Resources Branch, Bethesda, Maryland, USA, for part of the recombinant human IL-2 used for in vivo studies. JM was supported by the Intramural Research Program; National Heart, Lung, and Blood Institute; NIH.

ASJC Scopus subject areas

General Medicine

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10.1172/jci.insight.152014

Cite this

Thomas, A., Sumughan, S., Dellacecca, E. R., Shivde, R. S., Lancki, N., Mukhatayev, Z., Vaca, C. C., Han, F., Barse, L., Henning, S. W., Zamora-Pineda, J., Akhtar, S., Gupta, N., Zahid, J. O., Zack, S. R., Ramesh, P., Jaishankar, D., Lo, A. S. Y., Moss, J., ... Le Poole, I. C. (2021). Benign tumors in TSC are amenable to treatment by GD3 CAR T cells in mice. JCI Insight, 6(22), Article e152014. https://doi.org/10.1172/jci.insight.152014

@article{64465679e3ac41cd9fce2420e90e6fb9,

title = "Benign tumors in TSC are amenable to treatment by GD3 CAR T cells in mice",

abstract = "Mutations underlying disease in tuberous sclerosis complex (TSC) give rise to tumors with biallelic mutations in TSC1 or TSC2 and hyperactive mammalian target of rapamycin complex 1 (mTORC1). Benign tumors might exhibit de novo expression of immunogens, targetable by immunotherapy. As tumors may rely on ganglioside D3 (GD3) expression for mTORC1 activation and growth, we compared GD3 expression in tissues from patients with TSC and controls. GD3 was overexpressed in affected tissues from patients with TSC and also in aging Tsc2+/- mice. As GD3 overexpression was not accompanied by marked natural immune responses to the target molecule, we performed preclinical studies with GD3 chimeric antigen receptor (CAR) T cells. Polyfunctional CAR T cells were cytotoxic toward GD3-overexpressing targets. In mice challenged with Tsc2-/- tumor cells, CAR T cells substantially and durably reduced the tumor burden, correlating with increased T cell infiltration. We also treated aged Tsc2+/- heterozygous (>60 weeks) mice that carry spontaneous Tsc2-/- tumors with GD3 CAR or untransduced T cells and evaluated them at endpoint. Following CAR T cell treatment, the majority of mice were tumor free while all control animals carried tumors. The outcomes demonstrate a strong treatment effect and suggest that targeting GD3 can be successful in TSC.",

author = "Ancy Thomas and Saurav Sumughan and Dellacecca, {Emilia R.} and Shivde, {Rohan S.} and Nicola Lancki and Zhussipbek Mukhatayev and Vaca, {Cristina C.} and Fei Han and Levi Barse and Henning, {Steven W.} and Jesus Zamora-Pineda and Suhail Akhtar and Nikhilesh Gupta and Zahid, {Jasmine O.} and Zack, {Stephanie R.} and Prathyaya Ramesh and Dinesh Jaishankar and Lo, {Agnes S.Y.} and Joel Moss and Picken, {Maria M.} and Darling, {Thomas N.} and Scholtens, {Denise M.} and Dilling, {Daniel F.} and Junghans, {Richard P.} and {Le Poole}, {I. Caroline}",

note = "Publisher Copyright: {\textcopyright} 2021, Thomas et al.",

year = "2021",

month = nov,

day = "22",

doi = "10.1172/jci.insight.152014",

language = "English (US)",

volume = "6",

journal = "JCI Insight",

issn = "2379-3708",

publisher = "American Society for Clinical Investigation",

number = "22",

}

Thomas, A, Sumughan, S, Dellacecca, ER, Shivde, RS, Lancki, N, Mukhatayev, Z, Vaca, CC, Han, F, Barse, L, Henning, SW, Zamora-Pineda, J, Akhtar, S, Gupta, N, Zahid, JO, Zack, SR, Ramesh, P, Jaishankar, D, Lo, ASY, Moss, J, Picken, MM, Darling, TN, Scholtens, DM, Dilling, DF, Junghans, RP & Le Poole, IC 2021, 'Benign tumors in TSC are amenable to treatment by GD3 CAR T cells in mice', JCI Insight, vol. 6, no. 22, e152014. https://doi.org/10.1172/jci.insight.152014

TY - JOUR

T1 - Benign tumors in TSC are amenable to treatment by GD3 CAR T cells in mice

AU - Thomas, Ancy

AU - Sumughan, Saurav

AU - Dellacecca, Emilia R.

AU - Shivde, Rohan S.

AU - Lancki, Nicola

AU - Mukhatayev, Zhussipbek

AU - Vaca, Cristina C.

AU - Han, Fei

AU - Barse, Levi

AU - Henning, Steven W.

AU - Zamora-Pineda, Jesus

AU - Akhtar, Suhail

AU - Gupta, Nikhilesh

AU - Zahid, Jasmine O.

AU - Zack, Stephanie R.

AU - Ramesh, Prathyaya

AU - Jaishankar, Dinesh

AU - Lo, Agnes S.Y.

AU - Moss, Joel

AU - Picken, Maria M.

AU - Darling, Thomas N.

AU - Scholtens, Denise M.

AU - Dilling, Daniel F.

AU - Junghans, Richard P.

AU - Le Poole, I. Caroline

PY - 2021/11/22

Y1 - 2021/11/22

N2 - Mutations underlying disease in tuberous sclerosis complex (TSC) give rise to tumors with biallelic mutations in TSC1 or TSC2 and hyperactive mammalian target of rapamycin complex 1 (mTORC1). Benign tumors might exhibit de novo expression of immunogens, targetable by immunotherapy. As tumors may rely on ganglioside D3 (GD3) expression for mTORC1 activation and growth, we compared GD3 expression in tissues from patients with TSC and controls. GD3 was overexpressed in affected tissues from patients with TSC and also in aging Tsc2+/- mice. As GD3 overexpression was not accompanied by marked natural immune responses to the target molecule, we performed preclinical studies with GD3 chimeric antigen receptor (CAR) T cells. Polyfunctional CAR T cells were cytotoxic toward GD3-overexpressing targets. In mice challenged with Tsc2-/- tumor cells, CAR T cells substantially and durably reduced the tumor burden, correlating with increased T cell infiltration. We also treated aged Tsc2+/- heterozygous (>60 weeks) mice that carry spontaneous Tsc2-/- tumors with GD3 CAR or untransduced T cells and evaluated them at endpoint. Following CAR T cell treatment, the majority of mice were tumor free while all control animals carried tumors. The outcomes demonstrate a strong treatment effect and suggest that targeting GD3 can be successful in TSC.

AB - Mutations underlying disease in tuberous sclerosis complex (TSC) give rise to tumors with biallelic mutations in TSC1 or TSC2 and hyperactive mammalian target of rapamycin complex 1 (mTORC1). Benign tumors might exhibit de novo expression of immunogens, targetable by immunotherapy. As tumors may rely on ganglioside D3 (GD3) expression for mTORC1 activation and growth, we compared GD3 expression in tissues from patients with TSC and controls. GD3 was overexpressed in affected tissues from patients with TSC and also in aging Tsc2+/- mice. As GD3 overexpression was not accompanied by marked natural immune responses to the target molecule, we performed preclinical studies with GD3 chimeric antigen receptor (CAR) T cells. Polyfunctional CAR T cells were cytotoxic toward GD3-overexpressing targets. In mice challenged with Tsc2-/- tumor cells, CAR T cells substantially and durably reduced the tumor burden, correlating with increased T cell infiltration. We also treated aged Tsc2+/- heterozygous (>60 weeks) mice that carry spontaneous Tsc2-/- tumors with GD3 CAR or untransduced T cells and evaluated them at endpoint. Following CAR T cell treatment, the majority of mice were tumor free while all control animals carried tumors. The outcomes demonstrate a strong treatment effect and suggest that targeting GD3 can be successful in TSC.

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ER -

Benign tumors in TSC are amenable to treatment by GD3 CAR T cells in mice

Abstract

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