Kenneth M. Brinkhous: Investigator, Teacher, Administrator, and Gene Therapist
Authors: Gilbert C. White, Harold R. Roberts
Journal: Molecular Therapy (2001)

It is rare that an individual is privileged to see a disease from beginning to end, to be a part of a scientific beginning, and to contribute to a cure. Kenneth Merle Brinkhous (Fig. 1), who died December 11, 2000, at the age of 92, had that unusual privilege. In a remarkably long and consistent scientific career that included the longest continuously running R01 from the National Institutes of Health, Brinkhous would make many contributions, receive many accolades, and would ultimately be nominated for the Nobel Prize. But what excited him the most in the end was the possibility that he might live to see a cure for hemophilia, a disease he helped to define more than half a century earlier, and, more importantly, that he might contribute to the development of that cure. He had a remarkable passion for knowledge and scientific endeavor that drove him to remain productive even into his ninth decade and which endeared him to a whole new field of research, gene therapy, and a whole new generation of researchers. Although hemophilia was first described in antiquity, it was the pioneering work of Brinkhous and his colleagues, among others, in the middle half of the past century that led to the molecular definition of hemophilia. His work in the area started in 1939 with his description of anti-hemophilic factor, the factor missing in hemophilia A (1Brinkhous K.M. A study of the clotting defect in hemophilia: The delayed formation of thrombin.Am. J. Med. Sci. 1939; 198: 509-516Crossref Google Scholar). Later, the International Committee on Nomenclature, which Brinkhous helped found, would rename anti-hemophilic factor by its roman numeral designation, factor VIII. One of his most important observations was the development of the partial thromboplastin time (PTT), a functional assay of blood coagulation which could be used to measure specific components of blood, including factor VIII and factor IX (2Langdell R.D. Wagner R.H. Brinkhous K.M. Effect of anti-hemophilic factor on one-stage clotting tests.J. Lab. Clin. Med. 1953; 41: 637-647PubMed Google Scholar). The development of an accurate assay for the measurement of clotting factor activity would have far-reaching effects. Now used in laboratories all over the world for diagnostic purposes and for monitoring anticoagulant therapy, the PTT was critical to the facile measurement of factor VIII and factor IX and, ultimately, to their purification and characterization. In a celebration of the 25th anniversary of the development of the PTT, the famed photographer, Karch, was commissioned to photograph Langdell, Wagner, and Brinkhous (Fig. 2). The first commercial concentrates of factor VIII for therapeutic use in hemophilia derived from work by Wagner, his long-term colleague, and Brinkhous using a glycine precipitate method for factor VIII purification that maintained factor VIII potency (3Webster W.P. Roberts H.R. Thelin G.M. Wagner R.H. Brinkhous K.M. Clinical use of a new glycine-precipitated antihemophilic fraction.Am. J. Med. Sci. 1965; 250: 643-651Crossref PubMed Scopus (28) Google Scholar, 4Brinkhous K.M. Shanbrom E. Roberts H.R. Webster W.P. Fekete L. Wagner R.H. A new high-potency glycine precipitated antihemophilic factor (AHF) concentrate.J. Am. Med. Assoc. 1968; 205: 613-617Crossref PubMed Scopus (71) Google Scholar). This product revolutionized the treatment of hemophilia, leading to home treatment and permitting safe joint and other surgery in hemophiliacs. It remained a standard for nearly 20 years until blood-borne viruses gained entry to the blood supply and became the impetus for the development of safer and more efficient purification procedures. Early on, Brinkhous and his colleagues recognized the importance of animal models in the study of hemophilia. Because of the importance of this model in gene therapy studies, the story of the start of this colony is of some interest (J. B. Graham, personal communication). Around 1946, Brinkhous received a telephone call from a veterinarian at Cornell University. He told Brinkhous the story of a District Attorney in Cortland, New York, who raised Irish setters. Some of the setters were not doing well and had developed joint disease. The veterinarian had performed an autopsy on one of the animals that died and found a large spinal hematoma. Based on the bleeding and the joint disease, it was suspected that this might be canine hemophilia and Brinkhous was contacted. After confirmation of the diagnosis and some negotiation with the breeder, Brinkhous agreed to purchase Nora and Lynn, two female carriers. The dogs were driven from New York to Chapel Hill and that was the start of the hemophilia dog colony. Further characterization of the dogs revealed that the canine disorder was very similar to the human disease (5Graham J.B. Buckwalter J.A. Hartley L.J. Brinkhous K.M. Canine hemophilia. Observations on the course, the clotting anomaly, and the effect of blood transfusions.J. Exp. Med. 1949; 90: 97-111Crossref PubMed Scopus (52) Google Scholar). Later, Brinkhous would also acquire animals with canine hemophilia B, canine factor VII deficiency, and porcine von Willebrand's disease. The animals with hemophilia were used extensively in the 1960s through 1980s in the development of factor VIII and IX therapeutic concentrates. In the late 1980s, Verma and his colleagues at the Salk Institute contacted Brinkhous about a collaboration on gene transfer experiments using the hemophilia B dogs. Their initial work demonstrated expression of retrovirus-mediated canine factor IX in skin fibroblasts from hemophilic dogs (6Axelrod J.H. Read M.S. Brinkhous K.M. Verma I.M. Phenotypic correction of factor IX deficiency in skin fibroblasts of hemophilic dogs.Proc. Natl. Acad. Sci. USA. 1990; 87: 5173-5177Crossref PubMed Scopus (89) Google Scholar). Subsequent work with a number of investigators utilizing the hemophilia dog colonies provided strong preclinical data that played an important role in approval of the ongoing clinical trials of gene transfer in hemophilia A and B. Thus, through his vision and foresight in developing and maintaining an animal model of hemophilia, Brinkhous' decision to bring Nora and Lynn to Chapel Hill in 1946 continues to impact hemophilia research. Another word or two regarding his NIH grant (P. Ganguly, personal communication). In the summer of 1947, Brinkhous received a letter (Fig. 3) informing him that his grant application, initially designated RG-978 and entitled “Clotting Defect in Hemophilia: Studies of a Strain of Hemophilic Dogs,” was recommended for approval. In those days, the NIH had just been created from the Public Health Service and the grant was actually submitted to the Surgeon General of the United States. The individual institutes, NHLBI, NCI, etc., did not exist in those days and the original grant designation RG stood for Research Grant. In the application, Brinkhous asked for $14,000. Interestingly, he did not request indirect funds and the NIH added 8% to his budget to cover indirect costs. The grant was reviewed by five individuals and their reviews were half a page in length or less, each. This grant, later redesignated HL01648, was to be continuously funded for the next 50 years and was to become the longest continuously funded grant in the history of the NIH. Through his long career, Kenneth Brinkhous was many things. He was a visionary departmental chair who built a strong research and clinical Department of Pathology, a historian, a mentor, a successful stock trader who taught a college level course on the market, a founder of the International Society on Thrombosis and Haemostasis, an organizer of numerous scientific conferences, a loving father and husband, an entrepreneur, and, in the end, a gene therapist. Most of all, he was a lifelong scientist who worked in the laboratory until the age of 91.