Project title

Stealth-engineered cell therapy for autoimmune disease in the kidney 

Supervisors


Background

Cellular therapy using genetically engineered immune cells revolutionised the treatment of leukaemia and is now impacting autoimmune diseases such as systemic lupus erythematosus (SLE).  In children with SLE, the kidneys are frequently affected, leading to inflammation called lupus nephritis.  Cellular therapy may offer a means of treating this before irreversible damage is done. 

Cells used to make cell therapy typically come from the patient themselves, which is both costly and inefficient.  Using cells from healthy donors is potentially much more efficient.  However, the patient’s immune system can reject the infused cells, limiting the duration of effective therapy.  The interplay between this therapy and the host immune system is complex, and we seek to develop strategies for prolonging the therapeutic window.

We have recently developed a new therapeutic platform that uses γδT cells to deploy therapeutic antibodies at the disease site, recruiting local immune cells to attack target cells (2).  Importantly, γδT cells do not cause Graft vs. Host Disease (GvHD), which means they can be safely obtained from an unrelated donor.  Now, we seek to protect them from immune rejection by the recipient.

Aims and objectives

  1. To evaluate the impact of anti-CD20 antibody-secreting γδT cells on allogeneic PBMC
  2. To develop genetic engineering constructs to slow or prevent the rejection of allogeneic gdT cells in this context
  3. To model the therapeutic time-course of these combinations with respect to activation and differentiation dynamics within the donor and host immune cells
  4. To model these interactions in developmentally appropriate 3D models of the human kidney, as a model of lupus nephritis

 

Methods

Healthy donor γδT cells will be engineered to secrete anti-CD20 opsonins with or without an engineered cytokine.  Their ability to kill CD20+ cells will be determined using cytometric assays.  The engineered γδT will be co-cultured with autologous or allogeneic PBMC, and their effects on “host” immune cell activation will be determined initially using flow cytometry.  These panels will then be integrated into a larger spectral cytometric panel, allowing simultaneous analysis of multiple immune compartments.

A series of modules aimed at reducing allo-rejection will be added to the expression cassette, such as Serpin-B9 and PD-L1.  The degree of protection conferred by each module and the resultant effect on CD20+ target clearance will be determined.

The final part of the project will involve the creation of 3D culture systems that mimic the extracellular matrix of a solid organ such as the kidney.  In this instance, we will investigate the ability of the engineered gdT cells to penetrate the 3D models and deliver anti-CD20 cytotoxicity. 

Timeline

  • Months 0-6 – Training in γδT engineering, cytometric analysis and immune functional assays.
  • Months 7-12 – Spectral cytometric panel optimisation and comparison to lower-parameter flow cytometry.
  • Months 13-20  – Design and mechanistic analysis of “stealth” constructs
  • Months 21-26  – Evaluation of γδT persistence, serial cytotoxicity and proliferation in an allogeneic culture context.  Begin development of a 3D culture system.
  • Months 27-32  – 3D culture assays
  • Months 33-36  – Complete data analysis, write-up.


References

  1. Hayday, A., Dechanet-Merville, J., Rossjohn, J. & Silva-Santos, B. Cancer immunotherapy by γδ T cells. Science 386, eabq7248 (2024).
  2. Fowler, D. et al. Payload-delivering engineered γδT cells display enhanced cytotoxicity, persistence, and efficacy in preclinical models of osteosarcoma. Sci Transl Med 16, eadg9814 (2024).


Who should students contact?

Jonathan Fisher (jonathan.fisher@ucl.ac.uk).

Research topic

Genetics, Paediatric surgery