Year 3

The transplantation of blood forming stem cells from one individual to another is widely used to treat patients with otherwise incurable cancers. Because such transplantations alter the recipient immune system in profound ways there are many other applications for this powerful form of therapy. The studies proposed in this grant focused on the use of blood stem cell transplantation for the purpose of immune tolerance induction. Tolerance induction in this setting means that transplantation of blood stem cells trains the body of a recipient to accept organs from same stem cell donor without the need for drugs to suppress their immune system. Blood stem transplantations can also reverse aberrant immune responses in individuals with autoimmune diseases such as childhood diabetes and multiple sclerosis.

In this project we sought to develop new ways to perform blood stem cell transplants to make the procedure safer and therefore more widely useable for a broad spectrum of patients. Transplants can be dangerous and sometimes fatal. Serious complications are caused by the toxic chemotherapy or radiation which are used to permit stem cells to engraft, and by a syndrome called graft-versus-host disease. Our research has aimed to replace the toxic treatments by testing novel reagents that more specifically target and eliminate the cells in recipients that constitute the barriers to stem cell engraftment. Furthermore, we perform transplantations of purified blood forming stem cells, and thus are able to avoid the problem of graft-versus-host disease which is caused by non-stem cell “passenger” immune cells in the donor grafts.

The proposal has four Specific Aims. Aims 1 and 2 focus on development of biologic agents that specifically target recipient barrier cells. Aims 3 and 4 propose testing the reagents and approaches developed in the first two aims in mouse models to induce tolerance to co-transplanted tissues and to cure animals with muscular dystrophy, Type 1 diabetes mellitus and multiple sclerosis. These aims have not changed in this reporting period.

Our prior reports highlighted our progress in Aim 2, which is now complete. Aim 2 focused on the identification and testing of an antibody directed against a molecule called CD117 present on surface of human blood stem cells. We demonstrated that this antibody can safely target and eliminate human blood stem cells in mice that had been previously engrafted with human cells. Based upon these studies we were awarded a CIRM Disease Team Grant, which will test this anti-human CD117 antibody in a clinical trial for the treatment of children with severe combined immune deficiency (SCID), also known as the “bubble boy” disease. Children with SCID are missing certain types of white blood cells (lymphocytes) so they cannot defend themselves from infections. Without a transplant, SCID patients usually die before the age of two. Our proposed clinical study has the potential to significantly improve the success of transplants for these patients. This clinical trial will be a first to test a reagent that specifically targets recipient stem cells to clear niche space and allow replacement therapy by healthy donor stem cells.

In the last year we have continued to make significant progress on Aims 1, 3 and 4. Aim 1 proposed to study how to improve blood stem cell engraftment using novel agents in mice that have intact immune systems. The anti-CD117 antibody discussed above works well in recipients that lack lymphocytes but not recipients with normal immune function. We have tested the anti-CD117 antibody in mice that lack more defined lymphocyte subsets to narrow down which lymphocyte type must be neutralized or eliminated. We have also tested novel reagents that inhibit the activity of specific immune cells and observed a stronger effect of the anti-CD117 antibody when co-administered with these reagents. For Aims 3 and 4, we have successfully achieved our goal of performing blood stem cell transplants that result in the stable mixing of blood cells between donor and recipients (called partial chimerism). For Aim 3, recipients are from a specialized mouse strain that models muscular dystrophy (MDX mice). We have transplanted purified skeletal muscle stem cells (SMSC) and observed engraftment of SMSC in MDX mice injected with genetically-matched SMSC. The next step is to test if co-transplants of blood stem cells plus SMSC from genetically mismatched donors will permanently engraft and expand in MDX recipients. For Aim 4, two mouse models are studied: (1) NOD mice which model childhood diabetes, and (2) mice that develop multiple sclerosis. We can successfully block the progression of disease in these animals with blood stem cell transplants. Our next steps are to apply the therapies developed in Aim 1 to these disease models. In the post-award period we will continue to carry out studies testing the novel approaches developed here in models of tolerance induction.