Ewing sarcoma partial regression without GvHD by chondromodulin-I/HLA-A*02:01-specific allorestricted T cell receptor transgenic T cells

ABSTRACT

Background: Chondromodulin-I (CHM1) sustains malignancy in Ewing sarcoma (ES). Refractory ES carries a dismal prognosis and patients with bone marrow (BM) metastases do not survive irrespective of therapy. We assessed HLA-A*02:01/CHM1-specific allorestricted T cell receptor (TCR) wild-type and transgenic cytotoxic (CD8⁺) T cells against ES.

Patients and Methods: Three refractory HLA-A2⁺ ES patients were treated with HLA-A*02:01/peptide-specific allorepertoire-derived (i.e., allorestricted) CD8⁺ T cells. Patient #1 received up to 4.8 × 10⁵/kg body weight HLA-A*02:01⁻ allorestricted donor-derived wild-type CD8⁺ T cells. Patient #2 received up to 8.2 × 10⁶/kg HLA-A*02:01⁻ donor-derived and patient #3 up to 6 × 10⁶/kg autologous allorestricted TCR transgenic CD8⁺ T cells. All patients were treated with the same TCR complementary determining region 3 allorecognition sequence for CHM1 peptide 319 (CHM1³¹⁹).

Results: HLA-A*02:01/CHM1³¹⁹-specific allorestricted CD8⁺ T cells showed specific in vitro lysis of all patient-derived ES cell lines. Therapy was well tolerated and did not cause graft versus host disease (GvHD). Patients #1 and #3 showed slow progression, whereas patient #2, while having BM involvement, showed partial metastatic regression associated with T cell homing to involved lesions. CHM1³¹⁹ TCR transgenic T cells could be tracked in his BM for weeks.

Conclusions: CHM1³¹⁹-TCR transgenic T cells home to affected BM and may cause partial disease regression. HLA-A*02:01/antigen-specific allorestricted T cells proliferate in vivo without causing GvHD.

KEYWORDS:

• Allorestricted T cells
• adoptive transfer
• Ewing sarcoma
• immunotherapy
• T cell receptor transgenic T cells

Disclosure of potential conflicts of interest

S. Burdach has an ownership interest in PDL BioPharma and holds intellectual property (EU and US patents) in gene expression analysis. D. Busch holds shares in Juno Therapeutics. The other authors declare no potential conflicts of interest.

Acknowledgments

We thank all patients and families for their participation and support. Anna Hochholzer is acknowledged for technical assistance, Wolfgang Uckert and Matthias Leisegang for supplying the MP-71 vector and Wolfgang Schwingerand Ernst-Christian Urbanfor referral of patient#1.

Funding

This work was supported by grants from the Wilhelm Sander-Stiftung (2006.109.1), Else Kröner–Fresenius–Stiftung (GR & SB; P31/08//A123/07), BMBF (SB, PROVABES 01KT1311), the Deutsche Kinderkrebsstiftung (GR & SB; DKS 2010.07) and the BMBF (GR, UT and SB, TranSarNet 01GM1104B) as well as the Cura Placida Children's Cancer Research Foundation. T.G.P.G. is supported by a grant from ‘Verein zur Förderung von Wissenschaft und Forschung an der Medizinischen Fakultät der LMU München (WiFoMed)’, the Daimler and Benz Foundation in cooperation with the Reinhard Frank Foundation, by LMU Munich's Institutional Strategy LMUexcellent within the framework of the German Excellence Initiative, the ‘Mehr LEBEN für krebskranke Kinder – Bettina-Bräu-Stiftung’, the Fritz-Thyssen Foundation (FTF-40.15.0.030MN), the Friedrich-Baur Foundation, and the German Cancer Aid (DKH-111886 and DKH-70112257).

Author contributions

UT, SS, AK, MT, DS, FB, OS, RAR, TGPG, and SB did the experiments. UT, SS, IE, AK, MT, DS, FB, OS, RAR, TGPG, GHSR, PHS, and SB analyzed the data. UT, SS, AK, DS, FB, TGPG, GSHR, and SB interpreted the data. SJ, AR, and UD performed the statistical analysis. DHB provided multimer technology. UT, SS, KG, ITvL, and SB fulfilled diagnoses, collected patient information and did the follow up. UT and SB conceived, designed, and supervised the analysis. UT, SS, PHS, and SB drafted and revised the report.