Fiaccadori, Valeria; (2022) Development of a CD30-directed CAR-T cell for the treatment of relapsed/refractory classical Hodgkin Lymphoma. Doctoral thesis (Ph.D), UCL (University College London).

Text PhD_Thesis_Valeria_Fiaccadori.pdf - Accepted Version Access restricted to UCL open access staff until 1 September 2025. Download (5MB)

Abstract

Relapsed/refractory classical Hodgkin Lymphoma (cHL) represents an unmet clinical need. CD30 is an ideal target for CAR-T cell therapy in cHL, as its expression is high on tumour cells, but limited on healthy tissues. A proportion of activated T cells upregulates CD30, but the extent, duration and consequences of this on CD30-CAR T cell therapy have not been elucidated. Responses in early phase trials employing CD30-CAR T cells are encouraging, but their duration needs to be improved. To develop an efficacious CD30-CAR T cell, first I studied 11 CD30-binders for binding kinetics, stability and epitope binding. I tested them in vitro as second generation, 41BB co-stimulated CARs for cytotoxicity, cytokine release and proliferation against CD30+ targets. Binder 2 was selected for the best combination of on-target cytotoxicity and IL2 secretion. I studied CD30 expression on T cells at several timepoints after single activation. Expression peaked between day +4 and +5. By day +11, <10% of non-transduced T-cells expressed CD30, compared to up to 50% of CD30-CARs. These cells showed higher expression of activation markers (CD69 and CD62L). To elucidate its impact on CD30-CAR T cells activity, I knocked out CD30 on T cells using CRISPR-Cas9. I tested different timelines to incorporate editing in my in vitro CARs manufacturing and showed that editing non activated T cells was feasible, effective and did not impair subsequent activation or transduction. When tested in vitro, though, CD30 KO CD30-CARs did not show a statistically significant advantage compared to their non-edited counterpart and they achieved inferior tumour control In vivo, in an immunocompromised model. This works generated an efficacious second generation CD30-CAR, which will be used as backbone for further engineering and translation into clinic. While CD30 expression is high and persistent in CD30-CAR T cells, it does not appear to impact significantly on their activity in vitro and in vivo.

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