Harnessing native fat-residing stem cells for bone regeneration

Harnessing native fat-residing stem cells for bone regeneration

Funding Type: 
Early Translational II
Grant Number: 
TR2-01821
Award Value: 
$5,359,076
Disease Focus: 
Bone or Cartilage Disease
Stem Cell Use: 
Adult Stem Cell
Cell Line Generation: 
Other
Status: 
Active
Public Abstract: 
Like most tissues of the body, bone possesses a natural regenerative system aimed at restoring cells and tissues lost to natural cell aging, disease or injury. These natural regenerative systems are complex combinations of cell growth factors and support structures that guide and control the development of specialized bone stem cells. However, the regeneration process may still fail, for multiple reasons. For instance, the degree of skeletal injury may be so great that it overwhelms the natural regenerative capacity. Alternatively, the natural regenerative capacity may be defective; this is exemplified by osteoporosis, a frequent condition affecting post-menopausal females and elderly males and females. Osteoporotic individuals have severe declines in stem cell numbers (10-fold decrease from age 30 to 80) and stem cell function (tendency of stem cells to turn into fat rather than bone cells with age), leading to bone loss and “fragility fractures” that typically would not occur in persons with normal stem cell number and function. Thus, there is a tremendous need for therapies to increase the number and function of endogenous adult stem cells with the potential to build new bone. One option is to introduce so called mesenchymal stem cells (MSC) from the patient to bone repair sites. However, significant hurdles to autologous MSC use include the need for 2-3 week culture times to isolate MSC before application. Moreover culturing introduces infectious and immunogenic risks from prolonged exposure to animal products and cancerous risks from cellular gene changes in culture. In addition, once isolated, MSC require appropriate growth factor stimulation to form bone. Finally, MSC isolated from patient tissues such as fat or bone marrow are heterogenous and of undetermined composition—making growth factor dosing and conformance with FDA regulations for defining target product identity, purity, and potency more difficult. To circumvent these problems, we have identified and purified the cells at the origin of human MSC. We have termed these perivascular stem cells (PSC) because they are natively localized around all arteries and veins, forming the key cellular component of the natural regenerative system. In a significant breakthrough, we are able to isolate these cells within hours from adipose tissues in sufficient numbers for therapy without the need for culture. This realizes the possibility of harvesting and implanting stem cells during the same operative period. In another breakthrough, we have identified a potent growth factor NELL-1 that potently amplifies the ability of PSC to form bone and vascular structures. This has led to the development of our target PSC+NELL-1 product, which effectively stimulates and augments the body’s natural bone regenerative system by providing all the components (stem cells, growth factor, and allograft bone support structure) necessary to “jump start” as well as maintain the function of bone stem cells.
Statement of Benefit to California: 
This FDA oriented proposal focuses on crucial preparatory work required before IND-enabling preclinical studies on our Developmental Candidate for bone formation and regeneration. Our Developmental Candidate provides a complete package of stem cells, bone growth factors, and scaffold to build an optimized microenvironment to “jump start” bone formation in normal and impaired bone healing conditions. We have generated very promising preclinical data on our Developmental Candidate’s superior bone formation and regeneration efficacy. In addition to its significant impact on health care, this highly multi-disciplinary project by our team has many near-term and long-term benefits to the State of California. 1. Besides direct health costs, musculoskeletal injuries and diseases are the leading cause of work-related and physical disability in the United States. Hard working Californians are responsible for California’s annual gross domestic product of $1.8 trillion, which rank our state among the top eight largest economies in the world. By promoting the repair of both normal and healing-impaired bone in a safe and effective manner, our mature technology will reduce the loss of work productivity at the front end, reduce work disability costs, and reduce the loss of state income tax. 2. Local osteogenic stem cells decline with age (from 1/10,000 in newborns to 1/250,000 by age 30 and 1/2,000,000 by age 80), leading to osteoporosis, poor bone quality, and fragility fractures. In 1998, the health care burden for osteoporosis exceeded $2.4 billion in California alone. A whopping 64% of the $2.4 billion was caused by hip fracture. If our Development Candidate is successful in healing existing fractures in impaired bone, it may also translate to therapies to prevent fractures in impaired bone. This will significantly reduce the long-term health care burden for California’s public health insurance program. 3. This project directly adds jobs at [REDACTED] and at the California-based companies that are involved in this project. 4. This project will produce intellectual property that is owned by t [REDACTED]. Our team has a track record of attracting out of state private investment to invest in California and of procuring supplies and equipment from strategic California-based companies. 5. This mature project is precisely the type of cutting-edge, multi-disciplinary stem cell project that Californians imagined when they approved proposition 71 in 2004. The establishment of CIRM has transformed the research infrastructure at [REDACTED], increased our ability to recruit world class stem cell scientists, and attracted the attention of superb scientists from other disciplines to this new field. Working together, our team has compiled an impressive list of accomplishments and we are confident in our abilities to take this project to IND submission in a timely fashion. Funding of this project will fulfill the promise of proposition 71.
Progress Report: 

Year 1

Background: unsuccessful bone repair Most organs can regenerate cells lost to ageing, exposure to adverse conditions, ephemeral lifespan, disease or injury. Regenerative systems are complex combinations of growth factors and anchorage molecules which support, guide and control the maturation of specialized stem cells. The regenerative process may still fail, for multiple reasons. Bone fractures will sometimes not heal spontaneously because parts of the broken bone do not join anymore, or because bone regeneration itself is impaired, like in osteoporosis, a frequent condition that affects post-menopause females and elderly people of both genders. A novel class of bone-healing stem cells Since specialized bone-forming cells have not been identified, which stem cells could be used for the cell therapy of bone fractures? We have identified and purified novel stem cells, localized around blood vessels and therefore named perivascular stem cells (PSCs). We have also demonstrated that Nell-1, a potent osteogenic growth factor we have developed, efficiently turns PSCs into bone cells. In 2011, we have been granted support by the California Institute for Regenerative Medicine to develop a product to heal critically fractured bones in osteoporotic – or not – patients or to perform spine fusion in order to correct skeletal defects. PSCs will be purified from the patient’s own fat tissue, a well-documented, rich source of these cells, and embedded in a biocompatible scaffold in the presence of the bone forming growth factor. The resulting compound, inserted at the site of the fracture, will provide bone-forming stem cells 1-precisely characterized in terms of origin and identity; 2- derived from the patient, hence not rejected; 3- not cultured, therefore similar to their natural counterparts. Progress after one year of research supported by CIRM We are now close to the end of the first year of our CIRM supported studies. In this initial period we have, first, validated the stem cells of use with regard to stringent biologic criteria: how many of these cells can be purified from fat tissue? How viable and pure are these cells after the sorting process? Do stem cell numbers and quality depend on the age, sex and corpulence of the donor? Following analysis of about 80 distinct fat harvests (lipoaspirates) we have determined that robust bone forming PSC can be reliably purified, in sufficient numbers, from male and female donors in all age and weight ranges. We have also shown that PSC are at least as good, in terms of bone forming potential, than conventional mesenchymal stem cells (the candidate stem cells, so far, to be used for cell therapies of bone defects amd injuries), while being significantly superior regarding purity and safety. Transplantation experiments in mice and rats have revealed that the combination of PSC and the Nell-1 growth factor ensures the most efficient bone regeneration, but that PSC do not give rise to tumors, an important verification in any protocol involving transplantation. These experiments have also demonstrated that PSC stimulate the formation of new blood vessels, an essential requisite for efficient wound healing and tissue repair. Finally, we have embarked on the development of a larger animal model of bone regeneration, that will be used in the second and third years of the projects. In summary, we have reached all the milestones and met the deadlines planned in the original project. The next year will see the further validation of the protocol and the beginning of the development of a stem cell/scaffold/growth factor combination product in the perspective of trials in human patients.

Year 2

Our research group has identified in the human body a novel class of stem cells endowed with strong osteogenic potential, which means capacity to produce bone. These stem cells are attached to the outer aspect of blood vessels, and therefore present in all organs. Our ultimate goal is to use these stem cells in human beings, to perform for instance spine fusion, a procedure used to treat spine deformities (scoliosis) or faulty vertebrae, or to heal complicated bone fractures, especially in patients suffering from osteoporosis, a condition where spontaneous bone repair is compromised. We shall harvest stem cells from the patient’s own abdominal adipose (fat) tissue, a rich source of these cells which is also easy and safe to collect. Stem cells will be purified on an automated machine, named fluorescence activated cell sorter, and immobilized on an engineered biocompatible scaffold in the presence of a novel osteogenic growth factor, that is a sort of hormone which stimulates stem cell development into bone. We have won an Early Translational grant (2011-2014) from the California Institute for Regenerative Medicine to develop this project to the stage where the first clinical trials can be engaged into. In the first year, we had fully validated the stem cell population of use on a large number of human adipose samples and clearly documented the availability, efficiency and safety of these cells independently of the age, sex and weight of the donor. We had also initiated experiments in mice and rats which have been pursued during the second year, finally demonstrating unequivocally the outstanding bone forming ability of these cells in the presence of the appropriate scaffold and osteogenic hormone. We have also, during this 2nd year, developed a large animal model of stem cell mediated bone regeneration which will be indispensable prior to the inception of human treatments. Finally, we have refined the physicochemical properties of the scaffold for optimal osteogenic factor availability and bone growth. In conclusion, all milestones and deadlines included in this grant have been met and our team is steadily progressing toward the medical “translation” of these novel stem cells.

Publications

© 2013 California Institute for Regenerative Medicine