Robert A. Good - US grants

Short-lived autoimmune-prone mice which develop relatively early in life all of the diseases of aging, e.g., amyloidosis, autoimmunities, vascular diseases, coronary disease, immunodeficiency which renders them susceptible to infection, hyalinizing renal disease associated with high levels of circulating immunocomplexes and immunocomplex injuries, and a high frequency of malignancy, have a genetic fault that may be correctable by cellular engineering. We propose that the life span short-lived autoimmune-prone mice might be greatly prolonged and development of diseases of aging prevented if we can transplant marrow to these short-lived autoimmune-prone mice relatively early in life without producing graft-versus-host reaction (GVHR). This feat would require transplanting marrow to introduce genetically distinct stem cells. The stem cells should come from a strain of mice genetically programmed for long life, and late immunologic disorganization. Such a donor should be resistant to development of autoimmunity, and the marrow transplant should not produce GVHR.

A. SPECIFIC AIMS 1. To study the comparative influences of dietary calorie (energy restriction) selective fat restriction and selective carbohydrate restriction on cellular immunologic abnormalities that develop in autoimmune-prone mice and with aging in long-lived autoimmune-resistant mice. 2. To investigate the influence of dietary calorie (energy) restriction, selective dietary fat restriction, and selective carbohydrate restriction on development of B lineage precursor cells in autoimmune-prone NZB and long-lived autoimmune-resistant mice. 3. To determine the influences of dietary calorie (energy) restriction, selective dietary fat restriction, and selective carbohydrate restriction on decline of B lineage precursor cells in autoimmune-prone NZB and long-lived autoimmune-resistant mice. 4. To study the influence of dietary calorie (energy) restriction, selective fat restriction or selective carbohydrate restriction on endocrine metabolism and rate of DNA synthesis in the pyloric pits, bone marrow, spleen and thymus of mice from each of the several autoimmune-prone strains. 5. a. To determine whether autoimmune processes are involved in development of occlusive coronary vascular disease in autoimmune-prone mice. b. To determine whether total dietary calorie (energy), high fat or high carbohydrate intake are involved in development of coronary vascular lesions in autoimmunity-prone mice. 6. To determine whether autoimmunity-resistant, long-lived mice given a diet which will make possible development of coronary vascular lesions and atherosclerosis that occurs in short-lived, autoimmune-prone mice fed a high calorie, high saturated fat diet. For most of the experiments the mice will be individually caged, fed daily, the diets weighed, orts weighed and food intake recorded on each diet. The mice are weighed weekly and representative mice are sacrificed at 3-4 mo intervals for immunological study of lymph node and spleen cells, circulating immune complexes and serum. Studies of auto-antibodies in estrus cycles and of the influence of diets on hormone levels will be sampled at regular intervals with immuno-histochemical studies of kidney, heart, blood vessels, liver and spleen and histological study and chemical and histological analysis of heart, coronary vessels, aorta and aortic vessels will be made.

These investigations will analyze the effectiveness of lectin fractionated haploidentical parental or sibling marrow transplants in treatment of severe combined immunodeficiency or acute leukemia in man. If these investigations prove favorable, this approach will be extended to treatment of other highly lethal hematological and immunodeficiency diseases of man. Bone marrow transplantation in mice will also be investigated as a means to introduce resistance genes for the prevention of AKR leukemia or against the development of Potter myelomas in inbred mice and to prevent and treat autoimmune disease in NZB, NZBx NZW and other autoimmune-prone mice.

A. SPECIFIC AIMS 1. To study the comparative influences of dietary calorie (energy restriction) selective fat restriction and selective carbohydrate restriction on cellular immunologic abnormalities that develop in autoimmune-prone mice and with aging in long-lived autoimmune-resistant mice. 2. To investigate the influence of dietary calorie (energy) restriction, selective dietary fat restriction, and selective carbohydrate restriction on development of B lineage precursor cells in autoimmune-prone NZB and long-lived autoimmune-resistant mice. 3. To determine the influences of dietary calorie (energy) restriction, selective dietary fat restriction, and selective carbohydrate restriction on decline of B lineage precursor cells in autoimmune-prone NZB and long-lived autoimmune-resistant mice. 4. To study the influence of dietary calorie (energy) restriction, selective fat restriction or selective carbohydrate restriction on endocrine metabolism and rate of DNA synthesis in the pyloric pits, bone marrow, spleen and thymus of mice from each of the several autoimmune-prone strains. 5. a. To determine whether autoimmune processes are involved in development of occlusive coronary vascular disease in autoimmune-prone mice. b. To determine whether total dietary calorie (energy), high fat or high carbohydrate intake are involved in development of coronary vascular lesions in autoimmunity-prone mice. 6. To determine whether autoimmunity-resistant, long-lived mice given a diet which will make possible development of coronary vascular lesions and atherosclerosis that occurs in short-lived, autoimmune-prone mice fed a high calorie, high saturated fat diet. For most of the experiments the mice will be individually caged, fed daily, the diets weighed, orts weighed and food intake recorded on each diet. The mice are weighed weekly and representative mice are sacrificed at 3-4 mo intervals for immunological study of lymph node and spleen cells, circulating immune complexes and serum. Studies of auto-antibodies in estrus cycles and of the influence of diets on hormone levels will be sampled at regular intervals with immuno-histochemical studies of kidney, heart, blood vessels, liver and spleen and histological study and chemical and histological analysis of heart, coronary vessels, aorta and aortic vessels will be made.

We propose to continue long-term efforts to perfect bone marrow transplantation (BMT) as a means to prevent or treat many different diseases. This series of systematic experimental investigations will explore further the usefulness of bone marrow or fetal liver cell transplantation and ultimately stem cell transplantation as an experimental approach to analysis, prevention or treatment of a number of diseases and disorders not yet effectively treated. First we will explore BMT as an approach to prevention or to causation of senility in mice. We will try to prevent senility associated disease or disorders, including amyloid deposition and arthritis or arthroses, in senility accelerated mice [SAM(P)] mice. Alternatively, congenic or allogeneic BMT or congenic or allogeneic neonatal liver transplantation will be tested as a means of introducing from SAM(P) into the congenic resistant SAM(R) mice the senility accelerating propensity. Similarly, BMT will be tested as a means of causing, preventing or treating senility-associated changes and amyloidosis when BMT donors share the major histocompatibility complex (MHC) but are allogeneic to the SAM(P) mice, e.g. CBA/H or C3H/He. Alternatively, we will attempt to use donors that differ from SAM(P) at MHC, e.g. C57B1/6. BMT will also be tested as a means of preventing or treating two new models of non-inflammatory coronary vascular diseases or atherosclerosis in inbred mice. We will compare the immunological consequences of long-term BMT induced chimeric state where BMT have either been performed from autoimmune-resistant donors to mice of autoimmune-prone strains or to mice of autoimmune-resistant strains. We will also ascertain whether it is possible to treat effectively by BMT advanced, putatively irreversible kidney disease associated with autoimmunity states. Experiments will attempt to employ BMT from MHC-matched, MHC-mismatched or haploidentical donors to treat effectively advanced kidney disease by complete correction by BMT of the autoimmunity and immunologic dysregulation that led to the kidney disease in the first place. Finally, we will initiate experiments to evaluate the potential of stem cell preparations to reconstruct the entire hematopoietic and lymphoid systems and immunologic functions as an approach to correcting the genetically determined propensity to develop autoimmunities nad other diseases associated with aging and to analyze and influence the abnormal accumulations of Ly1+ B lymphocytes that occur in these autoimmune states.

This project has as its primary focus to achieve basic understanding of the influence of undernutrition without malnutrition -- chronic energy intake restriction (CEIR) which greatly increases life span and health span of rats and of mice of long-lived, and of short-lived autoimmune- prone and cancer-prone strains. We are attempting especially to analyze the mechanisms which underlie prevention by CEIR of retrovirus-induced mammary adenocarcinoma, certain autoimmunities, vascular diseases, malignancies and immunodeficiency disorders that develop with aging in mice of certain genetically short-lived strains, as well as the immunodeficiency associated with aging in mice of some long-lived strains. Particularly, we have discovered that CEIR down-regulates (tunes down) many forms of what we call vegetative and reproduction associated cellular proliferation -- while at the same time CEIR up-regulates cellular proliferation that is associated with liver regenerative repair following partial hepatectomy. Our experiments will attempt to analyze at the molecular level in one representative long-lived and in one representative short-lived strain and in one strain of mice in which females develop mammary adenocarcinoma in high frequency, how CEIR influences expression and production of growth factors that in turn influence and control cell proliferation in mammary gland epithelium, skin and esophageal and intestinal epithelium. We will also analyze how CEIR influences the expression and production of complete hepatic growth factors and a new augmentor of liver regeneration and how these actions influence hepatic cellular regeneration after partial hepatectomy. It is our view that the influence of CEIR on these crucial vegetative and adaptive proliferations play crucial roles in the action of CEIR to prolong life, prolong the span of health and prevent development of diseases which occur with aging such as cancers and autoimmunities.