PART TWO OF THREE Subject: RFAs Vol. 18, No. 27, August 11, 1989, Part 2 (final) Potential applicants interested in obtaining further information may write or call: Stanley L. Slater, M.D. Director, Geriatric Research and Training Program National Institute on Aging Room 5C27, Building 31 National Institutes of Health Bethesda, Maryland 20892 Telephone: (301) 496-6761 RFA AVAILABLE: NIH-89-HL-09-H MODELS FOR HYPERTENSION RESEARCH USING TRANSGENIC ANIMALS P.T. 34; K.W. 0755020, 0715115, 1002002 National Heart, Lung and Blood Institute* Application Receipt Date: January 16, 1990 PURPOSE The Division of Heart and Vascular Diseases of the National Heart, Lung, and Blood Institute (NHLBI), invites grant applications to develop and use transgenic animal models to study basic molecular, biochemical, cellular and physiological mechanisms involved in the pathogenesis of hypertension. Investigators skilled in transgenic techniques are encouraged to focus their efforts on understanding hypertension, and members of the hypertension research community are encouraged to collaborate with individuals skilled in the development and use of transgenic animals. BACKGROUND Cardiovascular disease, the leading cause of death in the United States, results in about 1 million deaths annually. Approximately 58 million Americans are afflicted with hypertension and this contributes significantly to cardiovascular death and disability. Hypertension exacts a large social and economic toll on the United States, and requires a substantial amount of health resources for its clinical management. Research aimed at understanding the molecular mechanisms involved in the etiology and pathogenesis of hypertension should lead to the development of new preventive and therapeutic strategies with favorable effects on both the clinical and economic consequences of this disease. Elucidation of the molecular abnormalities responsible for hypertension is greatly complicated by the multifactorial nature of the disease that depends on genetically driven physiological components which interact with each other and with environmental factors in complex ways. Research progress is greatly hampered by the lack of animal models with well defined, systematic, genetic alterations that can be used to study normal and abnormal processes involved in blood pressure regulation. New experimental techniques and strategies for developing and using transgenic animals will help to remove this major impediment. Indeed, the introduction of foreign genes into the reproductive cell line of mammals and the expression of those genes is one of the major technological advances in biology because it allows investigators to genetically manipulate animals at a level of detail and experimental control that has previously been unobtainable. Because the foreign genetic material is transmitted from generation to generation, it is possible to develop stable colonies of these genetically altered animals. Three techniques are used most commonly to introduce foreign genes into animals. The most frequently and successfully used approach is the direct microinjection of DNA into the pronucleus of a fertilized egg. The foreign genetic material becomes integrated into the germ line, and is subsequently transmitted from generation to generation. A second method for generating transgenic animals uses retroviral vectors. In this procedure recombinant DNA techniques are used to insert a gene into a non- replicating retrovirus, which is then used to infect embryonic cells of the developing animal. Although each method has unique advantages and problems, both suffer from a major limitation - the investigator cannot control where the foreign gene will become inserted in the host DNA, and hence cannot predetermine the defect or alteration in function. In addition, the inability to target a gene to a specific chromosomal location may interfere with the transplanted gene's ability to function, or may cause unwanted inactivation or activation of endogenous genes. The third strategy for developing transgenic animals seeks to overcome these obstacles by improving the ability to specify, or "target", the gene to be mutated. In this way, the investigator can modify or delete genes with greater precision. The technique, gene targeting by homologous recombination, has the potential to permit derivation of more precise animal models for human hereditary diseases and to provide higher levels of refinement and resolution in the study of the effects of individual genes on the whole animal. Other techniques are being developed to transfect cells to generate transgenic animals, such as the use of radiofrequency electrical pulses. Perhaps the most intriguing development is the recent report in which sperm cells incorporate foreign DNA in solution. The sperm are then used as vectors to introduce foreign genetic material into eggs. If reproducible, this technique could greatly reduce the time and expense to develop transgenic animals. The instrumental role that transgenic animals play in understanding fundamental biological phenomena is best illustrated by some of the important ways they are currently employed. Transgenic animals are being used: to study mechanisms responsible for tissue-specific and temporal- specific gene expression; to analyze the developmental activation of gene families; to identify neural and hormonal factors, and the mechanisms by which they regulate gene expression; to determine cell ancestry, cell location, and cell commitment during mammalian development; to assess the physiological function of particular cell types utilizing genetic ablation techniques; to generate new animal models of human diseases; to design and test novel therapeutic modalities; to develop new cell lines; to create animals for screening carcinogenic compounds; to produce clinically and economically important proteins; and to study the effect of altered levels of endogenous substances. OBJECTIVES AND SCOPE Transgenic animals offer new opportunities for the systematic dissection of the actions of genes in both health and disease. Investigators can unravel complex biological phenomena by studying selected events in whole organisms at the molecular, biochemical, cellular, and physiological levels. To date, the main applications of transgenic animals have focused on cancer, neurobiological diseases, and disorders of the immune system. Application of these technological advances to hypertension research has so far been limited. This RFA is designed to encourage the development and application of transgenic animal models to understand normal and aberrant development, structure, function, organization, regulation, and control of the cardiovascular system as it pertains to mammalian blood pressure regulation and the development of hypertension. Examples of topics are given below. The list is not complete; investigators are encouraged to study these or other topics that meet the objectives of this announcement. Representative areas of research include: 1. investigation of the molecular, biochemical, cellular, and physiological function of individual gene products involved in blood pressure homeostasis, and their interaction with other products under physiological and pathophysiological conditions. Methods to perturb physiological function may include gene targeting (to inactivate or alter endogenous genes) and insertional mutagenesis (to create mutant animals by inactivating endogenous genes). In addition, investigators may deliver new genes to alter levels of endogenous substances, to introduce substances not normally present, and to create animal models using human genes. 2. development of new therapeutic approaches 3. ascertainment of DNA sequences, trans-acting factors and mechanisms responsible for tissue-specific and temporal- specific gene expression of proteins playing a key role in blood pressure homeostasis and in the development of hypertension 4. identification and elucidation of the regulatory mechanisms of neural, hormonal, and other systems that are relevant to normal and abnormal blood pressure control 5. utilization of genetic ablation techniques to unravel the complex interactions of systems comprised of multiple cell types, such as the relationships between neurons and adjacent cells within the blood vessel 6. development and use of gene targeting strategies with larger mammals allowing investigators to use animals, such as the rat, that are more amenable to physiological manipulation than mice EXCLUSIONS While multidisciplinary approaches are encouraged, it is not the intent of this announcement to solicit applications for large studies encompassing a variety of independent projects, i.e., program projects. In general, funds will not be provided for the purchase and installation of expensive, new equipment. Awards under this announcement to foreign institutions will be made only for research of very unusual merit, need and promise, and in accordance with Public Health Service policy governing such awards. MECHANISM OF SUPPORT The support mechanism for this program will be the traditional, individual research grant. Although approximately $2.6 million in total costs for this program is included in the financial plans for fiscal year 1990, award of grants pursuant to this RFA is contingent upon receipt of funds for this purpose. It is anticipated that up to eight grants will be awarded under this program. The specific number to be funded, however, will depend on the merit and scope of the applications received and the availability of funds. Since a variety of approaches would represent valid responses to this announcement, it is anticipated that there will be a range of costs among individual grants awarded. Upon initiation of the program, the Division of Heart and Vascular Diseases will sponsor periodic meetings to encourage exchange of information among investigators who participate in this program, and to stimulate collaboration. Applicants should request ADDITIONAL TRAVEL FUNDS for a one-day meeting each year, most likely to be held in Bethesda, Maryland. Applicants should also include a statement in their applications indicating their willingness to participate in these meetings. Applicants, who will plan and execute their own research programs, are requested to furnish their own estimates of the time required to achieve the objectives of the proposed research project. Up to FIVE YEARS of support may be requested. At the end of the official award period, renewal applications may be submitted for peer review and competition for support through the regular grant program of the National Institutes of Health (NIH). It is anticipated that support for the present program will begin in July 1990. Administrative adjustments in project period and/or amount of support may be required at the time of the award. If collaborative arrangements involve sub-contracts with other institutions, the Chief, Blood Diseases Section, NHLBI Grants Operation Branch should be consulted regarding procedures to be followed (tel: 301-496-7255). All current policies and requirements that govern the research grant programs of the NIH will apply to grants awarded under this RFA. REVIEW PROCEDURES AND CRITERIA Review Method: All applications submitted in response to this RFA will be evaluated for scientific and technical merit by an initial review group, which will be convened for this purpose by the Division of Extramural Affairs, NHLBI. Upon receipt, applications will be reviewed for their responsiveness to the objectives of this RFA. If an application is judged unresponsive, the applicant will be contacted and given the opportunity to withdraw the application, or have it considered for the regular NIH grant program. If an application submitted in response to this RFA is similar to an application already submitted to the NIH for review, the applicant will be required to withdraw the pending application before the new one is accepted. Simultaneous submission of similar applications will not be allowed. Review Criteria: The factors to be considered in the evaluation of scientific merit of each application will be similar to those used in the review of traditional research project grant applications, including: 1. the novelty, originality and feasibility of the approach and the adequacy of the experimental design 2. the competence of the principal investigator and collaborators to accomplish the proposed research, and the commitment and time they will devote to the project 3. the suitability of the facilities to perform the proposed research, including laboratories, instrumentation and data management systems 4. the appropriateness of the requested budget for the work proposed 5. adequate plans for interaction and communication of information and concepts among investigators involved in collaborative studies. METHOD OF APPLICATION Letter of Intent: Prospective applicants are asked to submit a letter of intent to apply to this RFA. This letter should include the names of any participating institutions and all investigators, together with a descriptive title. Such a letter of intent is not binding and it will not enter into the review of any application subsequently submitted, nor is it a necessary requirement for application. Letters of intent are requested solely for planning purposes. The NHLBI Staff will not provide responses to such letters. Letters of intent to apply to this RFA should be received no later than December 1, 1989, and should be addressed to: Dr. Charles L. Turbyfill Review Branch Division of Extramural Affairs National Heart, Lung and Blood Institute Westwood Building, Room 553 NIH, Bethesda, MD 20892 Format for Applications: Submit applications on form PHS-398 (revised 10/88), the application form for the traditional NIH research project grant. Copies of this form are available in the applicant institution's office of sponsored research, or may be obtained from the following: Office of Grants Inquiries Division of Research Grants Westwood Building, Room 449 NIH, Bethesda, MD 20892 Use the conventional format for research project grant applications and ensure that the points identified in the section above on "Review Procedures and Criteria" are fulfilled. To identify the application as a response to this RFA, CHECK "YES" on item 2 of page 1 of the application and enter the title "Models for Hypertension Research Using Transgenic Animals" and enter the RFA number NIH-89-HL-09-H in the space provided. THE RFA LABEL FOUND IN THE FORM PHS-398 APPLICATION KIT MUST BE AFFIXED TO THE BOTTOM OF THE FACE PAGE OF THE ORIGINAL COMPLETED APPLICATION FORM PHS-398. FAILURE TO USE THIS LABEL COULD RESULT IN DELAYED PROCESSING OF YOUR APPLICATION SUCH THAT IT MAY NOT REACH THE REVIEW COMMITTEE IN TIME FOR REVIEW. Application Procedure: Send or deliver the completed, signed application and four (4) complete photocopies to the following, making sure that the original application with the RFA label attached is on top: Division of Research Grants Westwood Building, Room 240 National Institutes of Health Bethesda, MD 20892** SEND TWO ADDITIONAL COPIES OF THE APPLICATION TO DR. CHARLES TURBYFILL AT THE ADDRESS LISTED UNDER "LETTER OF INTENT". IT IS IMPORTANT TO SEND THESE TWO COPIES AT THE SAME TIME AS THE ORIGINAL AND FOUR COPIES ARE SENT TO THE DIVISION OF RESEARCH GRANTS, OTHERWISE THE NHLBI CANNOT GUARANTEE THAT THE APPLICATION WILL BE REVIEWED IN COMPETITION FOR THIS RFA. Applications must be received by January 16, 1990. An application not received by this date will be considered ineligible. Timetable: Letter of Intent December 1, 1989 Application Receipt Date January 16, 1990 Review by National Heart, Lung and Blood Advisory Council May 25-26, 1990 Anticipated Award Date July 1, 1990 Inquiries: Inquiries regarding this announcement may be directed to the following: Dr. Stephen C. Mockrin Deputy Chief Hypertension and Kidney Diseases Branch Division of Heart and Vascular Diseases National Heart, Lung and Blood Institute Federal Building, Room 4C10 NIH, Bethesda, MD 20892 (Tel: 301-496-1857) *The programs of the Division of Heart and Vascular Diseases, National Heart, Lung, and Blood Institute, are identified in the Catalogue of Federal Domestic Assistance, Number 13.837. Awards will be made under the authority of the Public Health Service Act, Section 301 (42 USC 241) and administered under PHS grant policies and Federal regulations, most specifically 42 CFR part 52 and 45 CFR part 74. This program is not subject to the intergovernmental review requirements of Executive Order 12372 or to review by a Health Systems Agency RFA: 89-HL-12-H MOLECULAR AND CELLULAR EMBRYOLOGY OF HEART MUSCLE P.T. 34; K.W. 0705015, 1002017, 1002004, 1002008 National Heart, Lung And Blood Institute * Application Receipt Date: January 16, 1990 PURPOSE The Division of Heart and Vascular Diseases (DHVD) of the National Heart, Lung, and Blood Institute (NHLBI) invites grant applications for five years of support for research focused on cardiac myogenesis and on the perturbations of normal developmental processes which result in malformations of the myocardium. Of particular interest are studies of the lineage and developmental pathways of the cells of the heart chambers and the regional specialized tissues within them. BACKGROUND At the present time, there is only limited knowledge of how the heart achieves its size and shape and specifies regional function. Morphogenesis of the heart, like other organs, is a complex process which involves coordination of cell migration, division and death as well as differentiation and adhesion. Furthermore, this process is not solely intrinsic to the cells, but is the result of cellular and tissue interactions. In the earliest stages, endoderm provides information necessary for development of heart mesoderm. As development of the heart proceeds, signalling appears to occur between the developing outflow tract and other regions of developing myocardium. This is evidenced by the fact that outflow tract anomalies appear to influence development of ventricular myocardium and the formation of the cardiac chambers. Thus, for example, mitral or aortic valve dysplasia or aortic arch anomalies are often associated with malformation of the left ventricle. This solicitation is focused on research leading to elucidation of the mechanisms involved in cardiac muscle cell genesis, proliferation and differentiation. A number of questions need to be answered. What are the progenitors of cardiac myocytes? Does cell proliferation occur only in stem cells or can committed myocytes resume cell division in response to an appropriate stimulus? How is mitotic activity controlled in the developing heart? How is differentiation initiated and controlled? Early studies of cardiac morphogenesis showed that waves of mitotic activity travelled through heart mesoderm from its rostral end to its caudal end and that a major decline in mitotic activity occurred a few hours before striated myofibrils were first seen in the heart. Subsequently, sinoatrial cells were seen to replicate at a higher rate than that seen in ventricular cells. Moreover, when contraction was initiated in the cardiac tube, it occurred first on the right side. Thus, regional variations in time of differentiation and onset of function are observable in the developing heart. It has also been reported that in early chick embryos, development of cardiac beat rate is regulated by regional cues rather than by intrinsic properties of the cells. Thus, replication, differentiation, and the assumption of specialized function were shown to be subject to complex controls that involve both extracellular and intracellular mechanisms. Recent studies of the incorporation of bromodeoxyuridine into hearts of early chick embryos, demonstrating regional differences in rates of DNA synthesis, have confirmed these early observations. Computer graphic reconstruction of these results has shown regions of quiescent cells. These images leave little doubt that regional decreases in proliferation or DNA synthesis participate in cardiac morphogenesis through local restriction of tissue growth. It remains to be determined how tightly DNA synthesis is coupled to karyokinesis, ploidy, cytokinesis or regional differences in cytoplasmic volume. Such cytokinetics, as well as migrations or sheet movements of cells, may also influence the emerging shape of the definitive heart. There is also evidence that some regions of quiescent cells may represent specialized tissues such as atrial natriuretic peptide storage sites and conducting tissue. Understanding of myogenesis and cardiac cell proliferation and differentiation must involve a study of a multiplicity of mechanisms which initiate and regulate gene expression. Research to date has largely focused on the role of oncogenes in modulating phenotypic expression in cardiac myocytes. These genes may be more relevant to cardiac hypertrophy than to developmental processes. This solicitation will, therefore, foster investigations which seek to identify candidate proteins and their genes associated with cardiac morphogenetic and developmental processes. One approach to the dissection of the various developmental processes is to establish a cardiac cell line which can be manipulated by various experimental procedures. Two attempts to utilize transgenic mice to obtain replicating myocytes resulted in hypertrophy of the atria. Cells from these atria will proliferate in culture. Continuous cell division of mammalian ventricular cells which maintain differentiated phenotypic properties has not yet been achieved. However cardiac muscle cells from amphibians will undergo cytokinesis 15-20 days after wounding the heart. In this case, both atrial and ventricular cells showed evidence of cell division. Another possible source of a cell line is a pluripotent stem cell line which might be induced to differentiate into cardiac myocytes. Cell lineage analysis in the developing heart is an important area for investigation, and much remains to be uncovered. It is not always the case that the temporal sequence of differentiation and the final generation of cell diversity proceed in a uniform, step-wise fashion. For example, in the rat retina, some of the cell types are generated in the embryo, but most rod photoreceptors are generated in the newborn pup. Cell lineage analysis shows that a retinal progenitor cell can give rise to at least two different cell types right up to the stage of its final division. Similar studies are needed in the mammalian heart, taking advantage of new approaches to marking cells, for example, by gene transfer using replication-defective retroviral constructs. Other relevant areas include the roles of growth factors, cell-cell communications, cell adhesion molecules, substrate adhesion molecules, and position of the cell within the developing organ as factors in cell regulation. OBJECTIVES AND SCOPE Applicants are encouraged to submit applications for up to five years of support for studies relevant to the topic of this RFA. The proposed studies should have clearly stated hypotheses and be focused on a well-defined problem. Collaborative work may be proposed where appropriate. It may involve individuals from within the investigator's institution or it may be interinstitutional. PROPOSED RESEARCH Research topics could include: o Elucidation of cell lineages of cardiac myocytes, with emphasis on cells of the various chambers and those with specialized function such as impulse generation and conduction o Studies of cell and cell-sheet migrations which are relevant to the assembly of the myocardium o Identification and elucidation of the roles of growth factors and genes involved in the morphogenetic and developmental process o Studies of the roles of spacial and temporal factors in cardiac development o Studies of the roles of intercellular communication in the developmental process o Elucidation of the mechanisms whereby physiological function can modulate the organization of the myocardium o Studies of the roles of angiogenesis or vascularization and the delivery of oxygen, cytokines and growth factors on growth and maturation of heart muscle These topics are provided as examples only: investigators are strongly urged to consider other relevant projects and approaches not mentioned above. EXCLUSIONS Applications that propose only the development of a permanent myocyte cell line without using it to examine aspects of heart muscle development will not be considered relevant to this RFA. Similarly studies focused exclusively on cells other than myocyte progenitors and myocytes will not be considered relevant. Applicants may request equipment; however funds will not be provided for major, expensive items or for outfitting a laboratory. MECHANISM OF SUPPORT The support mechanism for this program will be the traditional, individual research project grant. Although the financial plans for fiscal year 1990 include $1,500,000 for the total costs of this program, award of grants pursuant to this RFA is contingent upon receipt of funds for this purpose. The specific amount to be funded will, however, depend on the merit and scope of the applications received and on the availability of funds. Since a variety of approaches would represent valid responses to this announcement, it is anticipated that there will be a range of costs among individual grants awarded. It is not the intent of this announcement to solicit applications for large studies encompassing a variety of independent projects, i.e., program projects. If collaborative arrangements involve sub-contracts with other institutions, the Section Chief, DHVD, NHLBI Grants Operations Branch should be consulted regarding procedures to be followed (tel: 301-496-7255). Upon initiation of the program, the Division of Heart and Vascular Diseases will sponsor annual meetings to encourage an exchange of information among investigators who participate in this program. In the preparation of the budget for the grant application, applicants should REQUEST ADDITIONAL TRAVEL FUNDS for a two-day meeting each year to be held in Bethesda, Maryland. Applicants should also include a statement in their applications indicating their willingness to participate in such meetings. Applicants, who will plan and execute their own research programs, are requested to furnish their own estimates of the time required to achieve the objectives of the proposed research project. Up to FIVE YEARS of support may be requested. At the end of the official award period, renewal applications may be submitted for peer review and competition for support through the regular grant program of the National Institutes of Health (NIH). It is anticipated that support for the present program will begin on July 1, 1990. Administrative adjustments in project period and/or amount of support may be required at the time of the award. All current policies and requirements that govern the research grant programs of the NIH will apply to grants awarded in connection with this RFA. Awards in connection with this announcement will be made to foreign institutions only for research of very unusual merit, need, and promise, and in accordance with Public Health Service policy governing such awards. REVIEW PROCEDURES AND CRITERIA Review Method All applications responding to this RFA will be reviewed for scientific and technical merit by an initial review group, which will be convened by the Division of Extramural Affairs, NHLBI. Upon receipt, applications will be reviewed for their responsiveness to the objectives of this RFA. If an application is judged unresponsive at this stage, the applicant will be contacted and given an opportunity to withdraw the application or to have it considered for the regular research grant program of NIH. If an application submitted in response to this RFA is substantially similar to a research grant application already submitted to NIH for review, the applicant will be required to withdraw either the pending application or new one. Simultaneous submission of substantially similar applications will not be allowed. Review Criteria The factors to be considered in the evaluation of each application will be similar to those used in the review of traditional research project grant applications. The major factors to be considered in the evaluation of applications will include: 1. The scientific merit of the proposed projects, including the originality and feasibility of the approach, and the adequacy of the experimental design; 2. The competence of the investigators to accomplish the proposed research goals, their commitment, and the time they will devote to the program; 3. The adequacy of facilities for performance of the proposed research including the laboratory facilities, the proposed instrumentation and, when needed, the data management systems; 4. The integration of any interdisciplinary components into a coherent enterprise with adequate plans for interaction and communication of new information and concepts among the collaborating investigators; 5. The appropriateness of the budget for the proposed program METHOD OF APPLYING Letter of Intent Prospective applicants are asked to submit a letter of intent that includes identification of any other participating investigators and institutions, together with a descriptive title. The NHLBI requests such letters only for the purpose of providing an indication of the number and scope of applications to be received and, therefore, usually does not acknowledge their receipt. A letter of intent is not binding, and it will not enter into the review of any application subsequently submitted, nor is it a necessary requirement for application. This letter of intent, which should be received no later than November 30, 1989, should be sent to: Dr. Charles Turbyfill Review Branch, Division of Extramural Affairs National Heart, Lung, and Blood Institute Westwood Building, Room 553 National Institutes of Health Bethesda, Maryland 20892 Format for Applications Submit applications on form PHS 398, (revised 10/88) the application form for the traditional research project grant. This form is available in the applicant institution's office of sponsored research or business office. Use the conventional format for research project grant applications and ensure that the points identified in the section on "Review Procedures and Criteria" in this announcement are fulfilled. Be sure to observe the new page limitation requirements. To identify the application as a response to this RFA, CHECK "YES" on Item 2 of page 1 of the application and enter the title "Molecular and Cellular Embryology of Developing Heart Muscle" and the RFA number 89-HL-12-H. IN ADDITION, BE SURE TO ATTACH THE RFA LABEL FROM THE APPLICATION KIT TO THE BOTTOM OF THE FACE PAGE OF THE ORIGINAL APPLICATION AND TO PLACE THIS ON TOP OF THE FOUR COPIES TO BE MAILED TO THE DIVISION OF RESEARCH GRANTS. FAILURE TO USE THIS LABEL COULD RESULT IN DELAYED PROCESSING OF YOUR APPLICATION SUCH THAT IT MAY NOT REACH THE REVIEW COMMITTEE IN TIME FOR REVIEW. Application Procedure Send or deliver the completed application and four (4) signed, exact photocopies to the following address, making sure that the original application with the RFA label attached is on top: Division of Research Grants Westwood Building, Room 240 National Institutes of Health Bethesda, Maryland 20892** SEND AN ADDITIONAL TWO (2) COPIES OF THE APPLICATION TO DR. CHARLES L. TURBYFILL AT THE ADDRESS LISTED UNDER LETTER OF INTENT. IT IS IMPORTANT TO SEND THESE TWO COPIES AT THE SAME TIME AS THE ORIGINAL AND FOUR COPIES ARE SENT TO THE DIVISION OF RESEARCH GRANTS. OTHERWISE THE NHLBI CANNOT GUARANTEE THAT THE APPLICATION WILL BE REVIEWED IN COMPETITION FOR THIS RFA. Applications must be received by January 16, 1990. An application not received by this date will be considered ineligible. Timetable Letter of Intent November 30, 1989 Application receipt date January 16, 1990 Review by National Heart, Lung and Blood Advisory Council May 25-26, 1990 Anticipated Award Date July 1, 1990 Inquiries Inquiries regarding this announcement may be directed to the program administrator: Constance Weinstein, PhD Division of Heart and Vascular Diseases National Heart, Lung, and Blood Institute Federal Building, Room 3C06 National Institutes of Health Bethesda, Maryland 20892 Telephone: (301) 496-1081 * This program is described in the Catalog of Federal Domestic Assistance number 13.837, Heart and Vascular Diseases. Awards will be made under the authority of the Public Health Service Act, Section 301 (42 USC 241) and administered under PHS grant policies and Federal regulations, most specifically 42 CFR Part 52 and 45 CFR Part 74. This program is not subject to the intergovernmental review requirements of Executive Order 12372, or to Health Systems Agency Review. RFA: 89-HL-10-H BASIC DEVELOPMENTAL BIOLOGY OF THE VESSEL WALL P.T. 34; K.W. 0705015, 0775000, 1002004 National Heart, Lung, and Blood Institute* Application Receipt Date: December 13, 1989 PURPOSE The Division of Heart and Vascular Diseases (DHVD) of the National Heart, Lung, and Blood Institute (NHLBI) invites grant applications to support research studies on formation of the vessel wall during embryogenesis. Basic research is encouraged to address the genetic factors controlling phenotypic diversity of the cells of the vascular wall, to investigate factors that initiate and control cell proliferation and differentiation, and to study the effects of aberrations in these processes that may induce arteriosclerosis and other vascular diseases. BACKGROUND Investigations on the embryogenesis of the vessel wall and on factors initiating and controlling cell wall proliferation and differentiation will advance understanding of the vascular biology events with the hope that common mechanisms important to atherogenesis and a range of other vascular diseases can be described. Atherosclerosis is generally accepted as a reaction involving smooth muscle proliferation and extra- and intra- cellular lipid accumulation. Recent data suggest that atherogenesis begins early in life and that atherosclerotic lesions probably occur at "pre-existing" sites of intimal smooth muscle. Other studies have suggested that growth factors control smooth muscle proliferation or that the proliferative change begins with a focal developmental localization of smooth muscle cells into intimal cell masses. Yet, little is known as to how and when the intima of vessels form, or whether smooth muscle cells can continue to be derived from more primitive cell types even in adult life. Further, the relationship of smooth muscle cells to smooth muscle-like cells (e.g. pericytes) remains obscure. Even the cellular lineages of smooth muscle and endothelial cells are not clear. Recently, it has been reported that genetic hypertension in the rat is present either as a growth defect or as a very early abnormality of pressure control. At present, it is not known how the wall of resistance vessels is formed, and how it becomes remodeled to produce the thickened wall of the hypertensive microvasculature. Equally unknown is the role of endogenously derived growth factors and proteins, derived from other genes controlling cell growth, in the development of hypertension or in the increases in vascular mass. Whether these and other substances controlling growth arise in the smooth muscle cell itself or in interactions with other cells of the vessel wall is unknown. Vascular inflammation is another area where vascular developmental biology may be important for understanding vascular pathology. Many inflammatory diseases are specific for individual vascular beds. Advances in immunology and virology indicate that this specificity probably reflects regional specialization in the endothelium, either as an antigen-presenting cell or as a cell with an appropriate target for a virus or bacterial agent. It is not known, however, how these specializations arise or whether they represent fundamental properties of the local endothelium or adaptations to the local parenchymal environment. Embryologic studies suggest that the endothelium develops over a short period of time as a small number of cell aggregates, some of which form discrete blood islands. The fate of these primordia in the adult is unknown. Studies of endothelial cells from different vascular beds suggest that these cells, in culture, may have very different properties. If true, this would imply either that the primordia were different or that interactions with specific organ parenchyma had led to variation in genetic expression in the endothelium. The data from different vascular beds need to be confirmed and mechanisms controlling differentiation need to be explored at the molecular level. OBJECTIVES AND SCOPE There are a number of major important unresolved issues in vascular developmental biology: Recent embryologic studies suggest that smooth muscle cells in local organs may be derived as a result of interactions of invading vascular channels and the organ primordium. Mechanisms of induction of smooth muscle cell differentiation from more primitive cells are not known, or how smooth muscle cells from various vascular beds differ. There are working hypotheses to explore the mechanisms underlying vascular growth; e.g., a number of reports have shown that several polypeptides control the same cells which replicate in atherogenesis and other disease processes. Most of the work in this area, however, has emphasized growth factors derived from platelets or inflammatory cells. It seems unlikely that these exogenous sources of mitogens operate during development or in the hypertrophic response to high blood pressure. This strongly suggests that other unidentified mechanisms must be operative and should be explored. Similarly, recent work suggests that vessel wall cells themselves may function as the source of growth factors. It is intriguing to consider the possibility that endogenous growth factors may be responsible for vascular hypertrophy in hypertension or that vasoactive factors may also have trophic functions. Whether this occurs in vivo and how the production of such factors is controlled remain unclear. Mechanisms underlying differentiation of smooth muscle and endothelial cells and their possible interrelationships have been investigated. There would seem to be a relationship between the original formation of blood vessels and angiogenic responses later in life. Studies of co-cultured endothelium and smooth muscle cells suggest that these cells may interact to control each others' growth. There are reports that some, as yet undefined, endothelial factor may be able to reduce the expression of differentiated contractile proteins by smooth muscle cells, a phenomenon known as "modulation". It has been suggested that modulation is necessary in order for differentiated smooth muscle cells to replicate. However, it is not clear how the endothelial cells may control the recruitment and differentiation of smooth muscle and whether similar processes may be important in responses to injury in the adult vessel. Recently, specific molecules thought to be involved in angiogenesis have been identified and the genes cloned. However, little is known about how these molecules function during development, whether their primary effects are as mitogens, controlling replication, or as morphogens, controlling differentiation. Studies of the mechanisms of angiogenesis have generally focused on tumor angiogenesis. However, the issue of neo-vascularization is equally important in issues central to the vascular system, including collateral vessel formation in areas of infarction, invasion of atherosclerotic plaques by the vasa vasorum, recanalization of thrombosed vessels, and structural adaptation of the microcirculation to changes in blood flow. In summary, recent advances in molecular embryology have begun to permit identification of cell lineages and have begun to allow the identification of specific genes able to control cellular differentiation. However, little is known of how vessels become organized or about whether cells comprising vessels in different sites have common embryologic origins. An understanding of the embryology of vessel wall cells could provide information about the localization of atherosclerosis, inflammatory processes and other injuries to specific vascular beds. EXAMPLES OF RESEARCH PROJECTS This RFA is designed to encourage research on vascular biology events at a basic level of cellular and molecular biology with the hope that common mechanisms important to a range of vascular diseases can be described. Examples of topics that would be appropriate for this program are given below. The list is not to be regarded as complete or exclusive and other research areas proposed by applicants that meet the objectives of this program will be considered by the NHLBI. Representative areas of research might include the identification and elucidation of: o Factors controlling endothelial and smooth muscle cell growth during development, o Mechanisms controlling differentiation of the cells of the vessel wall from primitive mesenchyme, o Genetic factors controlling phenotypic diversity of the cells of the vascular wall, o Role of endothelial cells in smooth muscle proliferation, differentiation and "modulation". o Formation of vascular intimal cells and their relationship to medial smooth muscle cells and the localization of areas prone to atherosclerosis. o Role of developmental mechanisms in the ontogeny of the hypertensive and the atherosclerotic vasculature. EXCLUSIONS Large scale clinical studies or clinical trials are not deemed appropriate for support under this solicitation. Although the research topics may require the use of transgenic animals, it is not the intent of this RFA to solicit applications for the development of animal models. MECHANISM OF SUPPORT The support mechanism for this program will be the traditional, individual research grant. Although approximately $1.25 million in total costs for this program is included in the financial plans for fiscal year 1990, award of grants pursuant to this RFA is contingent upon receipt of funds for this purpose. It is anticipated that up to 8 grants will be awarded under this program. The specific amount to be funded, however, will depend on the merit and scope of the applications received and the availability of funds. Since a variety of approaches would represent valid responses to this RFA, it is anticipated that there will be a range of costs among individual grants awarded. While multidisciplinary approaches are encouraged, it is not the intent of this announcement to solicit applications for large studies encompassing a variety of thematic projects, i.e., program projects. If collaborative arrangements involve sub-contracts with other institutions, the Section Chief, DHVD, NHLBI Grants Operations Branch should be consulted regarding procedures to be followed (tel: 301-496-7255). Upon initiation of the program, the Division of Heart and Vascular Diseases will sponsor periodic meetings to encourage exchange of information among investigators who participate in this program. In the budget for the grant application, applicants should request ADDITIONAL TRAVEL FUNDS for a one-day meeting each year, most likely to be held in Bethesda, Maryland. Applicants should also include a statement in their applications indicating their willingness to participate in these meetings. Applicants, who will plan and execute their own research programs, are requested to furnish their own estimates of the time required to achieve the objectives of the proposed research project. Up to FIVE YEARS of support may be requested. At the end of the official award period, renewal applications may be submitted for peer review and competition for support through the regular grant program of the National Institutes of Health (NIH). It is anticipated that support for the present program will begin in July 1990. Administrative adjustments in project period and/or amount of support may be required at the time of the award. All current policies and requirements that govern the research grant programs of the NIH will apply to grants awarded under this RFA. Awards under this announcement to foreign institutions will be made only for research of very unusual merit, need and promise, and in accordance with Public Health Service policy governing such awards. REVIEW PROCEDURES AND CRITERIA Review Method: All applications submitted in response to this RFA will be evaluated for scientific and technical merit by an initial review group, which will be convened for this purpose by the Division of Extramural Affairs, NHLBI. Upon receipt, applications will be reviewed for their responsiveness to the objectives of this RFA. If an application is judged unresponsive, the applicant will be contacted and given the opportunity to withdraw the application, or have it considered for the regular NIH grant program. If an application submitted in response to this RFA is substantially similar to an application already submitted to the NIH for review, the applicant will be required to withdraw the pending application before the new one is accepted. Simultaneous submission of substantially similar applications will not be allowed. Review Criteria: The factors to be considered in the evaluation of scientific merit of each application will be similar to those used in the review of traditional research project grant applications, including: o the novelty, originality and feasibility of the approach and the adequacy of the experimental design o the competence of the principal investigator and collaborators to accomplish the proposed research, and the commitment and time they will devote to the project o the suitability of the facilities to perform the proposed research, including laboratories, instrumentation and data management systems o the appropriateness of the requested budget for the work proposed METHOD OF APPLICATION Letter of intent: Prospective applicants are asked to submit a letter of intent to apply to this RFA. This letter should include the names of any participating institutions and all investigators, together with a descriptive title. Such a letter of intent is not binding and it will not enter into the review of any application subsequently submitted, nor is it a necessary requirement for application. Letters of intent are requested solely for planning purposes. The NHLBI Staff will not provide responses to such letters. Letters of intent to apply to this RFA should be received no later than October 6, 1989, and should be addressed to: Dr. Charles L. Turbyfill Review Branch Division of Extramural Affairs National Heart, Lung, and Blood Institute Westwood Building, Room 553 NIH, Bethesda, MD 20892 Format for Applications: Submit applications on form PHS- 398 (revised 10/88), the application form for the traditional NIH research project grant. Copies of this form are available in the applicant institution's office of sponsored research, or may be obtained from the following: Office of Grants Inquiries Division of Research Grants Westwood Building, Room 449 NIH, Bethesda, MD 20892 Use the conventional format for research project grant applications and ensure that the points identified in the section above on "Review Procedures and Criteria" are fulfilled. To identify the application as a response to this RFA, CHECK "YES" on item 2 of page 1 of the application and enter the title, Developmental Biology of the Vessel Wall, and the RFA number, NIH-89-HL-10-H, in the space provided. THE RFA LABEL FOUND IN THE FORM PHS-398 APPLICATION KIT MUST BE AFFIXED TO THE BOTTOM OF THE FACE PAGE OF THE ORIGINAL COMPLETED APPLICATION FORM PHS-398. FAILURE TO USE THIS LABEL COULD RESULT IN DELAYED PROCESSING OF YOUR APPLICATION SUCH THAT IT MAY NOT REACH THE REVIEW COMMITTEE IN TIME FOR REVIEW. Application Procedure: Send or deliver the completed, signed application and four (4) complete photocopies of it to the following address, making sure that the original application with the RFA label attached is on top: Division of Research Grants Westwood Building, Room 240 National Institutes of Health Bethesda, MD 20892** SEND TWO ADDITIONAL COPIES OF THE APPLICATION TO Dr. CHARLES TURBYFILL AT THE ADDRESS LISTED UNDER "LETTER OF INTENT". IT IS IMPORTANT TO SEND THESE TWO COPIES AT THE SAME TIME AS THE ORIGINAL AND FOUR COPIES ARE SENT TO THE DIVISION OF RESEARCH GRANTS, OTHERWISE THE NHLBI CANNOT GUARANTEE THAT THE APPLICATION WILL BE REVIEWED IN COMPETITION FOR THIS RFA. Applications must be received by December 13, 1989. An application not received by this date will be considered ineligible. Timetable: Letter of Intent: October 6, 1989 Application Receipt Date: December 13, 1989 Review by National Heart, Lung and Blood Advisory Council: May 25-26, 1990 Anticipated Award Date: July 1, 1990 Inquiries: Inquiries regarding this announcement may be directed to the following: Momtaz Wassef, Ph.D. Deputy Chief Lipid Metabolism/Atherogenesis Branch Division of Heart and Vascular Diseases National Heart, Lung and Blood Institute Federal Building, Room 4A12 NIH, Bethesda, MD 20892 (Tel: 301-496-1978) * This program is described in the Catalog of Federal Domestic Assistance number 13.837, Heart and Vascular Diseases. Awards will be made under the authority of the Public Health Service Act, Section 301 (42 USC 241) and administered under PHS grant policies and Federal regulations, most specifically 42 CFR Part 52 and 45 CFR Part 74. This program is not subject to the intergovernmental review requirements of Executive Order 12372, or to Health Systems Agency Review.