Induction of immune tolerance with solid organ and vascular composite allografts is the Holy Grail for transplantation medicine. Induction of immune tolerance to mismatched grafts would obviate the need for life- long immunosuppression which is associated with serious adverse outcomes, such as renal failure, cancers and infections. Currently the most promising means of tolerance induction is through establishing a mixed chimeric state by transplantation of donor hematopoietic stem cells; however, with the exception of tolerogenic organs such as kidneys, the mixed chimerism approach has not achieved durable immune tolerance in preclinical or clinical trials with most solid organs or vascular composite allotransplants (VCA). Encouragingly, though, we have succeeded in achieving reduced immunosuppression in clinical trials of VCA using this approach. Mesenchymal stem (stromal) cells (MSC) are a potentially useful adjuvant to stem cell transplants (SCT) for promoting mixed chimerism as well as promoting complementary peripheral immunomodulatory functions. However, there are many unresolved issues to address before clinical translation of these promising therapeutic cells. A primary impediment is the source of MSC, which are rare in all tissues and require invasive procedures for procurement. Low abundance mandates extensive expansion in culture to generate sufficient numbers for human dosing. It has been observed in the clinical setting that the degree of expansion is negatively correlated with outcomes. Ossium Health has overcome this obstacle by identifying an abundant source of primary MSC associated with medullary bones of vertebral bodies obtained from deceased organ donors. These vertebral bone adherent MSC (vBA-MSC) are isolated by proteolytic digestion of bone fragments, following elution and cryopreservation of bone marrow (BM). Primary vBA-MSC are obtained at numbers that are 3 orders of magnitude higher than can be recovered from living donor iliac crest BM aspirates. A further advantage of vBA-MSC is they are matched to the donor, as opposed to third-party MSC, which enhances safety and potentially efficacy. Isolation and characterization of vBA-MSC from over 30 donors has demonstrated that the cells are no different than BM-MSC, but, because of their high numbers, unlike traditional BM-MSC, can be expanded to >5 billion cells with only 2 passages in culture. We hypothesize that donor-matched vBA-MSC will augment tolerance mechanisms of mixed chimerism with BM transplant as well as provide peripheral immunomodulatory functions to achieve durable tolerance for major histocompatibility complex mismatched solid organ and vascular composite tissue transplants. This hypothesis will be tested first in a murine orthotopic hind limb transplant VCA model and then in a murine heterotopic heart model. The hindlimb model studies will allow us to evaluate mechanisms of MSC immune tolerance due to the tolerogenic nature of BM-containing hind limbs. Durable grafts will be evaluated for T cell dynamics (especially memory T cells and regulatory T cells) and donor-specific immune tolerance with be confirmed with donor and third-party skin grafting. Information gained from this study will be used to perform similar studies in our heterotopic heart transplant model. If successful, the results of this study, combined with a plethora of MSC clinical trials as well as a long history of transplantation tolerance trials and our future Phase II studies to further define dosing parameters in small and large animal models, will provide compelling arguments to FDA for proceeding to clinical trials.
Keywords: vascular composite allotransplantation; solid organ transplantation; immune tolerance; immunomodulation, regulatory T lymphocytes
Public Health Relevance Statement: NARRATIVE Transplantation of solid organs, such as kidneys, liver and heart, are life saving procedures that are routinely performed. Relatedly, victims of traumatic injury who have lost limbs are candidates for limb transplants to restore quality of life and assist with re-integrating into the workforce. However, a negative consequence for transplant patients is the life-long requirement for drugs that suppress the immune system to prevent their body from rejecting the organ. These immunosuppression drugs are associated with severe medical issues such as cancer, kidney failure and infections. Therefore, new medical procedures that reduce or overcome the need for immunosuppressive drugs are greatly needed. Mesenchymal stem cells (MSC) have been shown to possess potent immune system modulating activities which has contributed to widespread testing in clinical trials for various diseases associated with immune system dysfunction. However, MSC are extremely hard to obtain and subsequently grow in the laboratory to generate sufficient numbers required to treat typical adult patients. Ossium Health has discovered very large numbers of MSC associated with the vertebrae of deceased donors who are also the source for most organs used for transplants. The company has developed novel methods to isolate these cells and shown that they expand quickly in the laboratory. A key advantage of these cells is they come from the same donor as the organ and, thus, pose little additional risk to the transplant recipient. We propose to test these MSCs for their ability to prevent rejection of transplanted limbs and hearts in mouse models that replicate the clinical scenarios. If successful, this new treatment modality may make organ and limb transplants safer, thereby, helping patients and reducing medical costs.
Project Terms: Address; Adjuvant; Adult; adverse outcome; Alloantigen; Allogenic; Allografting; allotransplant; Animal Model; Animals; Aspirate substance; Blood Vessels; bone; Bone Banks; Bone Marrow; Bone Marrow Transplantation; Bone Matrix; CD28 gene; CD3 Antigens; Cells; central tolerance; Chimerism; Chronic; Clinic; Clinical; clinical efficacy; clinical predictors; clinical translation; Clinical Trials; Complement; composite tissue transplantation; conditioning; Cryopreservation; Dendritic Cells; Digestion; Disease; Donor person; Dose; Engraftment; Evaluation; FOXP3 gene; Future; General Hospitals; Graft Rejection; Health; Heart; Heart Transplantation; Hematological Disease; Hematopoietic stem cells; Hindlimb; Human; Immune system; Immune System Diseases; Immune Tolerance; immunoregulation; Immunosuppression; In Vitro; in vivo; Individual; Infection; Kidney; Kidney Failure; kidney infection; Kidney Transplantation; Laboratories; Letters; Life; Limb structure; limb transplantation; Liver; Living Donors; Lymphocyte; Maintenance; Major Histocompatibility Complex; Malignant - descriptor; Malignant Neoplasms; Massachusetts; Medical; Medical Care Costs; Mesenchymal; Mesenchymal Stem Cells; Methods; Mixed Lymphocyte Culture Test; Modality; Modeling; mouse model; Mus; Nature; Non-Malignant; novel; Organ; Organ Donor; Organ Transplantation; Outcome; Patients; Peripheral; Pharmaceutical Preparations; phase 2 study; Population Dynamics; preclinical trial; prevent; Procedures; Property; Protocols documentation; Quality of life; Recording of previous events; Regimen; Regulatory T-Lymphocyte; Research Personnel; Resistance; Risk; Safety; Savings; Skin graft; Skin Transplantation; Solid; Source; spine bone structure; stem; Stem cell transplant; stem cells; Stromal Cells; Study models; success; T memory cell; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Therapeutic; Time; Tissues; transplant model; Transplant Recipients; Transplantation; transplantation medicine; Transplantation Tolerance; Traumatic injury; Universities; Upper Extremity; vertebra body; Vertebral Bone
