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The Orthopaedic Forum   |    
The Orthopaedic Genome: What Does the Future Hold and Are We Ready?
J. Edward Puzas, PhD; Regis J. O'Keefe, MD, PhD; Jay R. Lieberman, MD
View Disclosures and Other Information
J. Edward Puzas, PhD
Regis J. O’Keefe, MD, PhD
Department of Orthopaedics, University of Rochester School of Medicine and Dentistry, 601 Elmwood Avenue, Box 655, Rochester, NY 14642

Jay R. Lieberman, MD
Department of Orthopaedics, University of California at Los Angeles Medical Center, 10833 Le Conte Avenue, Los Angeles, CA 90095

Read at the Annual Meeting of the American Orthopaedic Association, Palm Beach, Florida, June 14, 2001

The authors did not receive grants or outside funding in support of their research or preparation of this manuscript. They did not receive payments or other benefits or a commitment or ageement to provide such benefits from a commercial entity. No commercial entity paid or directed, or agreed to pay or direct, any benefits to any research fund, foundation, educational institution, or other charitable or nonprofit organization with which the authors are affiliated or associated.

Presented at the Annual Meeting of the American Orthopaedic Association, Palm Beach, Florida, June 13-16, 2001.

The Journal of Bone & Joint Surgery.  2002; 84:133-141 
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The course of biomedical research and the practice of modern medicine have crossed into a new frontier—that of molecular genetics. The sequencing of the human genome will forever change the way that we view disease, and, in many cases, will point the way toward cures for diseases that we have not yet characterized. The disciplines of musculoskeletal research and clinical orthopaedics are part of this revolution. The following discussion highlights some of the recent advances in molecular and genetic biology, describes their possible application to the care of orthopaedic patients, and presents some of the social and ethical issues associated with the use of gene-altering therapy in humans.
 
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+Fig. 1:Diagrammatic representation of a gene. The promoter region controls whether or not the gene is expressed. Activators and repressors that bind to the promotor determine the level of expression. The coding region contains the information for protein production. The genetic information resides within the exons. Introns are "spacer" DNA sequences between the exons.
 
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+Fig. 2:Intracellular signaling cascade. Binding of a ligand to a receptor activates phosphorylating kinases. Subsequent activation steps eventually lead to the formation of an active transcription factor that controls gene expression in the cell’s genome.
 
Anchor for JumpAnchor for JumpTABLE I:  Diseases for Which Genetic Testing Is Available
Alpha-1-antitrypsin deficiency
Amyotrophic lateral sclerosis (ALS; Lou Gehrig disease)
Alzheimer disease
Ataxia telangiectasia (progressive brain disorder)
Gaucher disease (bone degeneration)
Inherited breast and ovarian cancer (BRCA-1 and BRCA-2)
Hereditary nonpolyposis colorectal cancer
Charcot-Marie-Tooth disease (CMT; loss of feeling in ends of limbs)
Congenital adrenal hyperplasia
Cystic fibrosis
Duchenne muscular dystrophy/Becker muscular dystrophy
Dystonia
Fanconi anemia group-C (FA; anemia, leukemia, skeletal deformities)
Factor V Leiden (FVL; blood-clotting disorder)
Fragile-X syndrome (fraX; leading cause of inherited mental retardation)
Hemophilia A and hemophilia B (HemA and HemB; bleeding disorders)
Huntington disease
Myotonic dystrophy
Neurofibromatosis type 1
Phenylketonuria
Adult polycystic kidney disease (APKD; kidney failure and liver disease)
Prader-Willi/Angelman syndromes (PWS/AS; decreased motor skills, cognitive impairment, early death)
Sickle-cell disease (SS; blood-cell disorder; chronic pain and infections)
Spinocerebellar ataxia, type-1 disorders (explosive speech)
Spinal muscular atrophy
Thalassemias
Tay-Sachs disease
 
Anchor for JumpAnchor for JumpTABLE II:  Relative Risk of Implant Loosening Related to Five Genetic Prognosticators
GeneFunctionRelative Risk
1Cytokine?0.85
2Cytokine inhibitor?2.2
3Chemokine?1.0
4Angiogenic factor?4.7
5Osteoclast activator10.2
The elucidation of the human genome is perhaps the single most meaningful project ever undertaken in biomedical research. The final outcome of this project will be the foundation from which new paradigms of curing disease will emerge.
The Human Genome Project was officially begun in 1990 through a coordinated effort of the United States Department of Energy and the National Institutes of Health. The project was slated to last for fifteen years but has been essentially completed in approximately ten years. A working database of DNA sequences is available now and, by 2003, the entire project should be complete. The stated goals of the project were (1) to identify all of the approximately 30,000 genes in the human genome, (2) to store the information in a form that would facilitate data analysis, (3) to develop more sophisticated sequencing methods, and (4) to address the ethical, legal, and societal issues that may arise from use of the information.
Knowledge of the human genome will impact every area of medicine and will revolutionize our field. We will gain a much more sophisticated understanding of the genetic determinants of disease, including an individual’s propensity for the development of diseases such as cancer, heart disease, and arthritis. Table I lists some of the diseases for which genetic testing has already commenced, and our ability to predict the likelihood of disease will continue to expand. Knowledge of the genome will aid in the development of powerful and specific pharmacologic agents and will increase our capacity to treat human disease. Drug design will be enhanced by the identification of specific gene targets, and pharmacologic agents will be synthesized to target the activity of a specific gene or protein. Gene therapy will become a reality in the future, and while it will undoubtedly be used for the treatment of diseases arising from genetic mutation, it will also probably be used for the treatment of developmental, degenerative, and traumatic conditions as well as cancer.
With the advent of the era of molecular medicine, important issues will confront orthopaedic surgeons, who will need to be conversant with the new technologies. There are moral, ethical, and medical considerations regarding genetic testing. Positive results of genetic tests that identify a predilection for cancer or other serious disorders only indicate the probability that the disease will develop. We know that many individuals who test positive will never have the disease and that individuals who show no predilection can still be affected. Compounded with the seriousness of the implications of a positive finding, errors in testing may have profound economic and/or social effects. Furthermore, the design of new drugs will raise issues regarding both safety and cost.
A genome is a compilation of genes that defines an organism or an individual. A gene is a sequence of nucleotides that contains a promoter region and a coding region (Fig. 1). The promoter region of a gene determines whether the gene will be expressed, and the coding region defines the sequence of amino acids in the resulting protein.
The orthopaedic genome is a constellation of thousands of genes that are directly involved in the genesis and maintenance of the musculoskeletal system. The orthopaedic genome includes gene families coding for connective-tissue matrix molecules (including both the biosynthetic and degradative aspects of their metabolism), the factors controlling skeletal development, bone and cartilage metabolism, and the intracellular and extracellular signaling pathways involved in the autocrine, paracrine, and endocrine control of skeletal function and repair. Notable discoveries in the study of the orthopaedic genome during the last decade have included the identification of parathyroid hormone-related protein (PTHrP)1, the bone morphogenetic proteins (BMPs)2, and the hedgehog family of molecules3 and the elucidation of their roles in skeletal development and fracture-healing. The identification of these primordial regulators of skeletogenesis has resulted in new approaches to clinical problems. For example, it soon may be possible to recapitulate the formation of skeletal tissues such as articular cartilage. Regeneration of new matrices rather than incomplete repair of existing cartilage should allow for better long-term results in the treatment of arthritis.
In the study of osteoclastogenesis, four proteins (receptor activator of nuclear factor-kappaB [RANK]; osteoprotegerin; osteoclast differentiation factor; and tumor necrosis factor-related activation-induced cytokine [TRANCE]) that regulate osteoclast formation and activity have recently been discovered4-7. RANK binds to a receptor on osteoclast precursor cells and effects their final differentiation into osteoclasts. Osteoprotegerin is a soluble decoy receptor that resembles RANK and inhibits osteoclast function8. Characterization of these molecules has provided alternative approaches for the control of bone resorption, which should lead to useful therapies for blocking bone loss in patients with osteoporosis, inflammation-mediated osteolysis, cancer, and other skeletal disorders.
Anabolic regulators of bone formation, including core binding factor alpha 1 (Cbfa1) and its gene (Cbfa1)9,10, have also been discovered. This gene codes for the transcription factor Cbfa1, which is both necessary and sufficient for the differentiation of cells into the osteoblast lineage and also enhances the differentiation of chondrocytes during endochondral bone formation. Thus, Cbfa1 is involved in osteoblast and chondrocyte differentiation and appears to play a key role in controlling the activity of these cells. Animals or humans deficient in Cbfa1 have arrested bone development, which leads to developmental abnormalities such as cleidocranial dysplasia.
Human DNA is comprised of forty-six chromosomes, which contain approximately 30,000 unique genes. Gene structure is highly conserved throughout nature, and the paradigm described in Fig. 1 holds not only for the animal kingdom but also for viruses, bacteria, and plants. Each individual gene is comprised of several characteristic elements, including a promoter, or regulatory region, and a transcriptional element, or coding region. Genes are important because they dictate cell and tissue function. Genes encode a sequence from which messenger RNA is transcribed, which, in turn, serves as a template for protein synthesis. Since the expression and function of specific proteins determine the character of cells, and ultimately tissues, the genetic sequence regulates both cell and tissue function.
Gene activation is dependent upon and regulated by the activation of transcription factors. In most cases, only a small portion of each gene directly codes for the protein sequence. The vast majority of the gene is comprised of sequences involved in the regulation of gene expression or of spacing sequences (introns). The promoter is the region of the gene that is directly involved in regulation. The promoter, typically up to 1000 bases in length, contains short DNA sequences that can bind to regulatory proteins called transcription factors. There are a large number of transcription factors; these factors recognize unique sequences of DNA, ranging from six to twelve bases in length, with high specificity. Transcription factors are able to recruit enzymes and other proteins necessary for messenger RNA synthesis. Each gene is regulated by multiple transcription factors that work both independently and in concert to regulate gene expression.
The portion of the gene that is responsible for coding messenger RNA contains both introns and exons. The exon sequences directly code for RNA, and the interval sequences between exons are referred to as introns. Intron sequences are enzymatically removed from newly transcribed RNA by a splicing mechanism that rejoins the exonic mRNA at specific sites. The mRNA is then translated on the ribosome to form specific proteins.
The binding of specific transcription factors to the promoter region of various genes controls the expression of these genes. Thus, the expression of a specific gene is dependent upon (1) the promoter sequence of that gene (i.e., the presence or absence of short sequences composed of six to twelve bases that permit specific transcription factors to bind to DNA) and (2) the presence of specific transcription factors within the cell. Some genes are necessary for basal function and are expressed in all cells. These genes are sometimes referred to as constitutively active genes, and their promoter region contains regulatory sites that bind to ubiquitously expressed transcription factors. Other genes are expressed in a much more restricted manner. The transcription factors that regulate these genes are similarly often selectively expressed. Thus, the Sox-9 gene, which induces type-II collagen, is expressed in mesenchymal cells undergoing chondrogenesis, and the Cbfa1 gene, which is necessary for bone formation and induces osteocalcin, is expressed exclusively in osteoblasts.
In many cases, transcription factors are present in cells but are in an inactive state. Many transcription factors are activated through the phosphorylation of serine or threonine amino acids located on these proteins. For some transcription factors, phosphorylation results in translocation from the cytoplasm to the nucleus, where they can bind to the promoter region of specific genes, leading to activation.
Transcription factor activation is the culmination of a series of phosphorylation events that comprise what is called a signaling pathway. Signaling pathways are composed of a series of sequentially activated kinases (Fig. 2), which are enzymes that add phosphate groups to target proteins. A paradigm that has emerged suggests that binding of a growth factor or other ligand to a cell membrane receptor leads to autophosphorylation and activation of the receptor. The activated receptor is then capable of phosphorylating a target kinase. A series of such events occurs until there is activation of a kinase that leads directly to transcription. Multiple, separate signaling pathways have been defined, including the protein kinase A, protein kinase C, Akt or protein kinase B, mitogen-activated kinase, and JAK/STAT (Janus Activated Kinase/signal transducer and activator of transcription proteins) signaling pathways.
During the last ten years, the production of large quantities of recombinant growth factors has had a great impact on the treatment of human diseases. An increasing number of recombinant growth factors have been approved for clinical use, and some of these factors have a role in the treatment of orthopaedic patients. For example, growth hormone and insulin-like growth factor-1 increase the longitudinal growth of bone in children deficient in these proteins11. Erythropoietin increases hematopoietic precursors and has been shown to decrease the need for perioperative transfusions in orthopaedic patients12. Bone morphogenetic proteins 2 and 7 are currently in phase-III clinical trials, and it is hoped that delivery of these growth factors to local sites will enhance reparative bone formation.
In spite of the successful clinical use of recombinant growth factors, there has been a shift away from such factors and toward signaling molecules as molecular therapeutic targets. This shift is due, in part, to the realization that the binding of growth factors to a membrane receptor is a complex event that usually activates several signaling pathways simultaneously. For example, the binding of BMPs to a specific receptor simultaneously activates three separate signaling pathways. Thus, activation of the Smad signaling pathway, which is responsible for BMP-signaling, is accompanied by activation of the mitogen-activated protein kinase (MAPK) and protein kinase-C signaling pathways13. As a consequence, BMP receptor-binding stimulates gene expression mediated by several separate transcription factors, including Smad, ATF-2 (activating transcription factor-2), and AP-1 (activating protein-1) transcription factors.
Therefore, recombinant growth factors, through receptor-mediated interactions, typically produce a large and heterogeneous response due to simultaneous activation of multiple signaling pathways and transcription factors. In some cases, these separate transcription factors can have disparate effects on target cells: one transcription factor may stimulate a desired cellular response (through activation of a specific set of genes), while another transcription factor may actually partially antagonize the response. Therefore, in theory, more exquisite control of cellular processes can occur with regulation at the level of intracellular signaling molecules. This is resulting in a conceptual shift from the use of recombinant growth factors toward the use of molecules targeting intracellular signaling pathways, a shift that will be accelerated by further progress in methods of gene delivery.
There are several strategies for targeting signaling pathways. One is the development of chemicals that inhibit or activate specific kinases or other enzymes. This strategy depends on knowledge of the three-dimensional structure of the protein. An example is the recent development and clinical use of a class of new molecules designed to inhibit the cyclooxygenase-2 enzyme. Another method is the direct delivery of genes that, once inside the cell, can use the intracellular apparatus to result in expression of an activated signaling protein. In contrast, there is the potential to genetically engineer genes that give rise to mutated proteins that can block specific signaling pathways. Other, more creative strategies being developed and investigated include the use of small RNAs or peptides that can inhibit the synthesis of specific proteins.
Methods of gene delivery have steadily improved, and the delivery of specific genes to cells in vitro has become relatively routine. The expression of genes is dependent upon the promoter that is used in a genetically engineered construct. Thus, a promoter that restricts expression to specific cell populations could be used. For example, when the type-X collagen promoter is used to drive expression, the target gene would be expressed only in hypertrophic chondrocytes and not in other cells. Alternatively, a promoter that includes a high level of expression in a large number of cells, such as the promoter for the viral large T antigen, could be used. Thus, introduced genes can have generalized or restricted patterns of expression. Furthermore, expression can be limited to specific cells at particular stages of differentiation, depending on the specificity of the promoter used in combination with the target gene.
The most challenging aspect of regulating gene expression has been the method used to deliver genetic material into the cell. DNA is highly charged and does not readily cross the cellular membrane; thus, several methods have been developed to insert DNA into the cell. The most common method in clinical trials has been the use of attenuated viruses that lack genes critical for viral replication. Thus, adenovirus-based gene therapy involves use of a replication-incompetent virus containing a target gene that encodes (expresses) a particular protein and is driven by a specific promoter.
Several strategies are being investigated to circumvent the immune response to adenovirus-based gene therapy. One method is the development of viral gene delivery methods in which there is no expression of viral proteins, and thus, no immune response. The adeno-associated viral vector, in particular, appears to have potential in this regard. However, this virus is much more difficult to produce and its ability to express target genes is less reliable than that of other viral delivery systems.
Yet another route for the introduction of specific proteins into cells takes advantage of a short twelve-amino-acid protein sequence (the so-called TAT protein), derived from the HIV genome. This peptide permits efficient intracellular uptake of recombinant proteins. The use of the TAT protein is a strategy for direct delivery of signaling proteins into cells, permitting modulation of intracellular signals that will alter the expression of those cells. Thus, recombinant proteins that contain this small TAT sequence can be delivered to cells in vitro or in vivo following local or systemic injection.
Successful tissue development and repair require the sequential expression of genes that modulate a complex set of events, including differentiation from primitive mesenchymal precursors, progressive maturation of cells, apoptosis or programmed cell death, and secretion of a specific and highly organized extracellular matrix. As the particular genes of the orthopaedic genome and the signals that regulate their temporal expression are elucidated, potential points of therapeutic intervention will become defined. These manipulations will affect a large number of orthopaedic conditions, including both reparative and degenerative conditions.
Thus, in the post-genomic era, when the sequences of all genes are known, the goal will be to find the subset of genes that are causative of or are directly related to a disease process. Knowing the sequence of genes alone is not the complete story; more important is their functional role. Functional genomics involves the knowledge of a specific gene’s role in both normal and pathological cell processes.
The key step, then, is to identify the small number of genes that are involved in a disease process from among the 30,000 genes that comprise the human genome. Efficient identification of important genes is facilitated by screening methods that permit the simultaneous analysis of a large number of genes. In one such method, called gene array, a large number of genes—as many as 10,000 to 20,000—are placed onto a small chip. The relative expression of the arrayed genes in different populations of cells can then be compared. This technology has led to breakthroughs in our understanding of some diseases. For example, the genes involved in the metastasis of malignant melanoma were recently defined14.
This methodology has potential applications in the study of musculoskeletal disease. One important target, in which the molecular events remain elusive, is osteoarthritis. This is among the most prevalent diseases in our society, affecting more than half of the individuals over the age of sixty-five and resulting in enormous health-care costs. Gene arrays that compare the patterns of gene expression in normal and osteoarthritic articular cartilage could lead to improved understanding of specific treatments for this disease.
There is a growing appreciation that the likelihood of human disease is dependent upon an individual’s genetic background. Different individuals can have diverse responses to the same stimulus, depending on the genetic determinants involved. For example, patients’ response to interleukin-1 (IL-1) correlates with their response to Helicobacter pylori infection15. In those with an extensive inflammatory reaction, the bacteria are eliminated but the gastric mucosa are damaged, leading to achlorhydria and an increased risk of gastric cancer. In contrast, in those who have a limited inflammatory reaction, the bacteria are not eliminated and gastric ulcers tend to develop. The basis of this finding lies in a single base difference in the promoter region of the IL-1 gene. Thus, while the region coding the IL-1 protein is the same in all individuals (i.e., all of our IL-1 protein sequences are identical), single base differences in the regulatory region result in differential levels of gene expression among different individuals, depending upon the sequences that they inherited. These important single base differences occur in nearly all genes and influence our propensity for disease. These differences are called single nucleotide polymorphisms, or SNPs.
It is likely that inflammatory conditions that affect the musculoskeletal system also depend upon specific patterns of gene expression. This has already been established for certain forms of inflammatory arthritis. Similarly, implant loosening, an inflammatory condition leading to osteolysis and failure, is also more likely in individuals with certain genetic backgrounds. Thus, a differential response to wear debris may explain why some prostheses loosen and others do not, and this genetic information may target future therapies toward patients at high risk for this complication.
Gene arrays permit exquisite molecular subtyping of diseases, such as cancer, that are heterogeneous in genetic expression but indistinguishable in clinical manifestation. The use of gene arrays potentially permits the widespread collection of genetic information on an individual basis. Just as each individual has a unique fingerprint, a unique genetic blueprint for each individual is possible and is likely to be defined in the future. With the growth of functional genomics, this information will provide important insight into the risk of a multitude of human diseases, including those of the musculoskeletal system. This development will markedly improve future medical care but has obvious ethical implications.
The following case study, a hypothetical assessment of a patient considering a total hip arthroplasty in the year 2010, is an adaptation of the "Shattuck Lecture—Medical and Societal Consequences of the Human Genome Project" by F.S. Collins16.
Assume that a sixty-year-old man with osteoarthritis of the left hip comes into your office. He has complete loss of the joint space and marked limitation of activity. He also has mild hypertension but is otherwise in good health. In addition to performing a complete history and physical examination, you scan the microchip on his insurance identification card and find that a "Genetic Index" screening for important diseases in men indicates that his relative risks for common cancers, heart disease, and osteoarthritis are 0.76, 0.53, and 4.20, respectively.
None of these genetic factors would predispose him to complications associated with a hip arthroplasty, so a further workup is performed. You order an "Arthroplasty Loosening Index" that examines the risk of implant loosening on the basis of the differential expression of genes known to be involved in prosthetic loosening. The relative risks associated with each of the five genes known to influence bone resorption around a prosthesis are depicted in Table II.
Considering all of the genes together results in an Overall Composite Loosening Index of 15.3. This means that the patient’s risk for the development of prosthetic loosening, based on his genetic background, is 15.3 times greater than that of the average person. After the patient’s age, weight, and activity level have been factored in, the Overall Composite Loosening Index indicates that he has a relative risk of 23.4. That is, he is 23.4 times more likely to undergo revision surgery for loosening of the implant within ten years than a low-risk patient is. Do you proceed with the operation?
Given the favorable Genetic Index score for important diseases and the relatively long life span expected for the patient, you proceed with the surgery. However, after the procedure you refer the patient to the Orthopaedic Molecular Therapy Unit for prophylaxis against prosthetic loosening.
Does this scenario represent fiction or reality? By the year 2010, it is likely to be a reality.
There are a number of ethical issues that require public debate, but we have chosen to focus on four areas of concern that influence both the field of medicine and society and that are particularly relevant to the care of patients with musculoskeletal problems. These ethical dilemmas include the following: (1) How will this new information regarding the human genome be used? (2) How will patient safety be maintained in an era in which we race to achieve new cutting-edge medical treatments that will benefit our patients but also will have the potential to provide huge profits for industry? (3) What safeguards need to be developed as partnerships between academia and industry continue to expand? and (4) Should advances in genetic medicine only be used to treat disease and genetic problems or should genetic enhancement of individuals be considered?
Our purpose is not to provide answers for these ethical dilemmas but to use this forum to stimulate public debate and to heighten awareness of the ethical challenges associated with this era of technological advancement.

Ethical Issues and the Human Genome

The identification of the human genome has received public attention worldwide. Scientists will use this information to identify the genes that are associated with particular diseases and inherited medical conditions and also to identify the signal transduction pathways that lead to various pathological conditions in order to develop effective treatment strategies. This information should enhance the efficacy of preventive medicine. There has been considerable interest in genetic testing over the past few years as genes associated with different cancers (e.g., breast and colon cancer) and other conditions have been identified. As our knowledge of the human genome increases and further links with various diseases are established, the demand for genetic testing will increase. At the present time, patients can undergo periodic screening examinations to diagnose a disease at an early stage, can change their lifestyle or diet in an attempt to delay the development of a disease or to limit its manifestations, and can even consider enrolling in experimental clinical trials. It is possible to envision a future scenario in which genetic information is used to develop gene therapies to treat a variety of diseases.
However, we will clearly need to develop strategies to handle this new information, given the obvious concerns about the improper use of such information by employers, health-care providers, and insurance companies. Will screening for the development of certain medical conditions become a routine part of pre-employment and pre-insurance testing? Should screening for genetic diseases be a routine aspect of a health-care evaluation? How should health-care professionals handle this information, particularly when the patient has a gene for a fatal disease for which there presently is no treatment? Does the physician have an obligation to inform other relatives, such as siblings and children, that they are at risk for the development of a fatal or seriously debilitating disease? What type of burden would this place on teenagers and young children if this information were revealed to them? Clearly, the potential for expanding the efficacy of preventive medicine is great, but these ethical dilemmas will have to be resolved by policymakers, clinicians, and, probably, the courts over time.

Patient Safety

Recent advances in genetic research will enable scientists to use gene transfer to treat diseases associated with genetic mutations (e.g., osteogenesis imperfecta) and a variety of other musculoskeletal problems, including rheumatoid arthritis; bone loss associated with fracture, nonunion, or revision total joint arthroplasty; and cartilage injury. This type of genetic manipulation is called somatic cell gene therapy17,18. Although investigators in this field are enthusiastic about the potential of using gene therapy to treat difficult clinical problems, the tragic death of Jessie Gelsinger, in an experimental gene-transfer trial at the University of Pennsylvania, demonstrates the inherent risk associated with the development of new treatment regimens. The association of this case with a general lack of protection for human subjects and the question of a financial conflict of interest highlights the potential ethical problems associated with the use of new technology19. Furthermore, the investigation of this death led to the identification of hundreds of unreported adverse events that have occurred among volunteers enrolled in different gene-transfer experiments. Interestingly, the involved researchers were in compliance with the requirements of the Food and Drug Administration even though the adverse events were not reported to the National Institutes of Health19. This tragedy has led to the development of an extensive educational effort regarding the importance of the protection of human research subjects and to the development of new regulations designed to limit adverse events in the future. Any clinical trial that is performed to test the efficacy of new drugs must provide maximum patient protection and informed consent. In addition, investigators must be wary of patients’ unrealistic expectations about the efficacy of new therapeutic regimens. This is of primary importance when treating patients with musculoskeletal conditions, many of whom are young and otherwise healthy. In most cases, the proposed therapy will not be life-saving and the goal will be to improve quality of life. Safety is more important than efficacy.
There is clear support for the development of new treatment strategies that employ either recombinant proteins, cytokines, or somatic cell gene therapy. However, the development of germ-line gene therapy is more controversial. This gene-transfer strategy would result in alteration of the genetic makeup of the sperm and ova, which would be passed on to succeeding generations18. A debate on the moral implications of germ cell gene transfer is beyond the scope of our discussion, but, as scientific breakthroughs occur, difficult ethical questions will arise and physicians should be ready to actively engage in debating these issues with policymakers and researchers.

Academia and Industry

Many of the most important advances in medicine that have been made in the past few decades have been the result of partnerships between academic institutions and industry. Such research collaborations can be extremely productive as long as academic freedom is maintained and maximum patient protection is ensured19,20. The potential for conflicts of interest is inherent in the practice of medicine, but it is magnified by the important role of pharmaceutical and biotechnology companies in the development of treatment modalities involving molecular medicine, particularly gene therapy. In addition, academic institutions have encouraged their faculty to develop biotechnology companies in which the institution has a substantial financial interest. Furthermore, the testing that is necessary in order to bring recombinant proteins or various gene-therapy strategies to market usually requires academic investigators to develop collaborations with industry because of the expense associated with clinical trials20. This may lead to a conflict of interest for the investigator because of the potential for huge profits associated with the successful development of these agents, particularly if the investigator owns stock in the company.
The potential for conflict of interest in these situations must be recognized. A study of pulmonologists showed not only that physicians’ prescribing patterns were affected by incentives offered by pharmaceutical companies but that clinicians were often unaware of the effect that such enticements had on their own behavior21. If physician behavior can be influenced by such small gifts as a pen or a free lunch, there is significant potential for a conflict of interest if more substantial financial gain is at stake. Investigators may overstate the potential efficacy of a particular treatment regimen to potential patients, and a truly objective analysis of the data might not be possible.
It is expected that an investigator would declare his financial interest during the informed-consent process. However, even the disclosure of potential conflict of interest to patients does not relieve the physician of the responsibility of making choices that are in the patient’s best interests. Patients may not fully understand the implications of a physician’s receiving financial compensation if the results of a particular trial are successful. Most patients enter the physician-patient relationship under the assumption that the physician will do what is in the patient’s best interest and may not suspect that financial remuneration may affect an important treatment decision22. Therefore, in these situations, disclosure of a potential conflict of interest to patients is probably not sufficient to safeguard the integrity of a clinical trial. Investigators with a direct financial interest in the outcome of the clinical trial should not be involved in the direction of the study, patient selection, or the informed-consent process20. Furthermore, medical journals should consider developing new policies so that potential conflicts of interest related to economic interests are scrutinized more closely.

Genetic Enhancement

Advances in molecular medicine have clearly captured the attention of all elements of our society. A recent article in Sports Illustrated entitled "Unnatural Selection" discussed the potential role of genetic engineering in producing a new breed of athlete who would exceed the present limits of human performance23. There is a general consensus that the development of gene-transfer strategies to treat medical diseases is acceptable but the use of such strategies for genetic enhancement is questionable. For example, the use of recombinant proteins or gene therapy to heal fractures, treat nonunions, and repair cartilage in order to enhance patients’ quality of life and to allow athletes to continue their activities seems quite reasonable. However, is it appropriate to use these techniques to enhance athletic performance by increasing muscle strength, oxygen transport, or overall body size? Evidently, athletes are already interested in using insulin-like growth factor to enhance muscle strength on the basis of findings in animal studies23. If our experience with anabolic steroids and erythropoietin provides any guidance, the abuse of growth factors to enhance athletic performance will become a reality if safeguards are not provided. In addition, physicians and the public need to be educated about the importance of using these growth factors in the correct way. It is well known that various growth factors can cause either cell proliferation or inhibition depending on the dosage used. In addition, growth factors may have different effects depending on the type of administration and the anatomical delivery site. Serious adverse events can be expected if these factors are used in the wrong way.
Will we eventually have a class of human beings who have been genetically altered to enhance athletic performance? Clearly, with the substantial potential for financial gain, this fantasy could become a reality. Public education strategies need to be developed, and governing bodies of various athletic organizations need to formulate policies defining both the proper and the illegal use of this new technology.
The "Principles of Medical Ethics" adopted by the American Academy of Orthopaedic Surgeons assert that the orthopaedic profession exists for the primary purpose of caring for the patient24. This requires that decision-making related to clinical care be focused on what is best for the patient and not what is best for the surgeon or a corporate sponsor24. As we enter this age of molecular medicine, we need to maintain our focus on patient protection and patient care. Orthopaedic surgeons need to participate in public education and public debate concerning the use of new molecular agents in order to enhance the care of our patients and to ensure that the appropriate safeguards are in place to protect them.
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Fuller K, Wong B, Fox S, Choi Y,Chambers TJ. TRANCE is necessary and sufficient for osteoblast-mediated activation of bone resorption in osteoclasts. J Exp Med,1998;188: 997-1001. 188997  1998  [PubMed]
 
Hsu H, Lacey DL, Dunstan CR, Solovyev I, Colombero A, Timms E, Tan HL, Elliott G, Kelley MJ, Sarosi I, Wang L, Xia XZ, Elliott R, Chiu L, Black T, Scully S, Capparelli C, Morony S, Shimamoto G, Bass MB,Boyle WJ. Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. Proc Natl Acad Sci U S A,1999;96: 3540-5. 963540  1999  [PubMed]
 
Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, Tomoyasu A, Yano K, Goto M, Murakami A, Tsuda E, Morinaga T, Higashio K, Udagawa N, Takahashi N,Suda T. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci U S A,1998;95: 3597-602. 953597  1998  [PubMed]
 
Simonet WS, Lacey DL, Dunstan CR, Kelley M, Chang MS, Luthy R, Nguyen HQ, Wooden S, Bennett L, Boone T, Shimamoto G, DeRose M, Elliott R, Colombero A, Tan HL, Trail G, Sullivan J, Davy E, Bucay N, Renshaw-Gegg L, Hughes TM, Hill D, Pattison W, Campbell P, Boyle WJ,et al. Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell,1997;89: 309-19. 89309  1997  [PubMed]
 
Ducy P. Cbfa1: a molecular switch in osteoblast biology. Dev Dyn,2000;219: 461-71. 219461  2000  [PubMed]
 
Karsenty G. Role of Cbfa1 in osteoblast differentiation and function. Semin Cell Dev Biol,2000;11: 343-6. 11343  2000  [PubMed]
 
Root AW, Kemp SF, Rundle AC, Dana K,Attie KM. Effect of long-term recombinant growth hormone therapy in children—-the National Cooperative Growth Study, USA, 1985-1994. J Pediatr Endocrinol Metab,1998;11: 403-12. 11403  1998  [PubMed]
 
Faris PM, Ritter MA,Abels RI. The effects of recombinant human erythropoietin on perioperative transfusion requirements in patients having a major orthopaedic operation. The American Erythropoietin Study Group. J Bone Joint Surg Am,1996;78: 62-72. 7862  1996  [PubMed]
 
Gallea S, Lallemand F, Atfi A, Rawadi G, Ramez V, Spinella-Jaegle S, Kawai S, Faucheu C, Huet L, Baron R,Roman-Roman S. Activation of mitogen-activated protein kinase cascades is involved in regulation of bone morphogenetic protein-2-induced osteoblast differentiation in pluripotent C2C12 cells. Bone,2001;28: 491-8. 28491  2001  [PubMed]
 
Clark EA, Golub TR, Lander ES,Hynes RO. Genomic analysis of metastasis reveals an essential role for RhoC. Nature,2000;406: 532-5. 406532  2000  [PubMed]
 
El-Omar EM, Carrington M, Chow WH, McColl KE, Bream JH, Young HA, Herrera J, Lissowska J, Yuan CC, Rothman N, Lanyon G, Martin M, Fraumeni JF Jr,Rabkin CS. Interleukin-1 polymorphisms associated with increased risk of gastric cancer. Nature,2000;404: 398-402. 404398  2000  [PubMed]
 
Collins FS. Shattuck lecture—medical and societal consequences of the Human Genome Project. N Engl J Med,1999;341: 28-37. 34128  1999  [PubMed]
 
Kaji EH,Leiden JM. Gene and stem cell therapies. JAMA,2001;285: 545-50. 285545  2001  [PubMed]
 
Szebik I,Glass KC. Ethical issues of human germ-cell therapy: a preparation for public discussion. Acad Med,2001;76: 32-8. 7632  2001  [PubMed]
 
Shalala D. Protecting research subjects—what must be done. N Engl J Med,2000;343: 808-10. 343808  2000  [PubMed]
 
Friedmann T. Medical ethics. Principles for human gene therapy studies. Science,2000;287: 2163 -5. 2872163  2000  [PubMed]
 
Orlowski JP,Wateska L. The effects of pharmaceutical firm enticements on physician prescribing patterns. There’s no such thing as a free lunch. Chest,1992;102: 270-3. 102270  1992  [PubMed]
 
Wenger NS,Lieberman JR. The orthopaedic surgeon and industry. Ethics and industry incentives. Clin Orthop,2000;378: 39-43. 37839  2000  [PubMed]
 
Swift EM,Yager D. Unnatural selection. Sports Illustrated,2001;94: 86-94. 9486  2001 
 
American Academy of Orthopaedic Surgery, Committee on Ethics 1996-1998. Guide to the ethical practice of orthopaedic surgery. 3rd ed. Rosemont, IL: American Academy of Orthopaedic Surgeons; 1998. p 10-11 
 

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Topics

Anchor for JumpAnchor for Jump
+Fig. 1:Diagrammatic representation of a gene. The promoter region controls whether or not the gene is expressed. Activators and repressors that bind to the promotor determine the level of expression. The coding region contains the information for protein production. The genetic information resides within the exons. Introns are "spacer" DNA sequences between the exons.
Anchor for JumpAnchor for Jump
+Fig. 2:Intracellular signaling cascade. Binding of a ligand to a receptor activates phosphorylating kinases. Subsequent activation steps eventually lead to the formation of an active transcription factor that controls gene expression in the cell’s genome.
Anchor for JumpAnchor for JumpTABLE I:  Diseases for Which Genetic Testing Is Available
Alpha-1-antitrypsin deficiency
Amyotrophic lateral sclerosis (ALS; Lou Gehrig disease)
Alzheimer disease
Ataxia telangiectasia (progressive brain disorder)
Gaucher disease (bone degeneration)
Inherited breast and ovarian cancer (BRCA-1 and BRCA-2)
Hereditary nonpolyposis colorectal cancer
Charcot-Marie-Tooth disease (CMT; loss of feeling in ends of limbs)
Congenital adrenal hyperplasia
Cystic fibrosis
Duchenne muscular dystrophy/Becker muscular dystrophy
Dystonia
Fanconi anemia group-C (FA; anemia, leukemia, skeletal deformities)
Factor V Leiden (FVL; blood-clotting disorder)
Fragile-X syndrome (fraX; leading cause of inherited mental retardation)
Hemophilia A and hemophilia B (HemA and HemB; bleeding disorders)
Huntington disease
Myotonic dystrophy
Neurofibromatosis type 1
Phenylketonuria
Adult polycystic kidney disease (APKD; kidney failure and liver disease)
Prader-Willi/Angelman syndromes (PWS/AS; decreased motor skills, cognitive impairment, early death)
Sickle-cell disease (SS; blood-cell disorder; chronic pain and infections)
Spinocerebellar ataxia, type-1 disorders (explosive speech)
Spinal muscular atrophy
Thalassemias
Tay-Sachs disease
Anchor for JumpAnchor for JumpTABLE II:  Relative Risk of Implant Loosening Related to Five Genetic Prognosticators
GeneFunctionRelative Risk
1Cytokine?0.85
2Cytokine inhibitor?2.2
3Chemokine?1.0
4Angiogenic factor?4.7
5Osteoclast activator10.2
Strewler GJ. The parathyroid hormone-related protein. Endocrinol Metab Clin North Am,2000;29: 629-45. 29629  2000  [PubMed]
 
Reddi AH. Bone morphogenetic proteins: from basic science to clinical applications. J Bone Joint Surg Am,2001;83 Suppl 1 (Pt 1): 1-6. 83 Suppl 1 (Pt 1)1  2001 
 
Juppner H. Role of parathyroid hormone-related peptide and Indian hedgehog in skeletal development. Pediatr Nephrol,2000;14: 606-11. 14606  2000  [PubMed]
 
Emery JG, McDonnell P, Burke MB, Deen KC, Lyn S, Silverman C, Dul E, Appelbaum ER, Eichman C, DiPrinzio R, Dodds RA, James IE, Rosenberg M, Lee JC,Young PR. Osteoprotegerin is a receptor for the cytotoxic ligand TRAIL. J Biol Chem,1998;273: 14363-7. 27314363  1998  [PubMed]
 
Fuller K, Wong B, Fox S, Choi Y,Chambers TJ. TRANCE is necessary and sufficient for osteoblast-mediated activation of bone resorption in osteoclasts. J Exp Med,1998;188: 997-1001. 188997  1998  [PubMed]
 
Hsu H, Lacey DL, Dunstan CR, Solovyev I, Colombero A, Timms E, Tan HL, Elliott G, Kelley MJ, Sarosi I, Wang L, Xia XZ, Elliott R, Chiu L, Black T, Scully S, Capparelli C, Morony S, Shimamoto G, Bass MB,Boyle WJ. Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. Proc Natl Acad Sci U S A,1999;96: 3540-5. 963540  1999  [PubMed]
 
Yasuda H, Shima N, Nakagawa N, Yamaguchi K, Kinosaki M, Mochizuki S, Tomoyasu A, Yano K, Goto M, Murakami A, Tsuda E, Morinaga T, Higashio K, Udagawa N, Takahashi N,Suda T. Osteoclast differentiation factor is a ligand for osteoprotegerin/osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci U S A,1998;95: 3597-602. 953597  1998  [PubMed]
 
Simonet WS, Lacey DL, Dunstan CR, Kelley M, Chang MS, Luthy R, Nguyen HQ, Wooden S, Bennett L, Boone T, Shimamoto G, DeRose M, Elliott R, Colombero A, Tan HL, Trail G, Sullivan J, Davy E, Bucay N, Renshaw-Gegg L, Hughes TM, Hill D, Pattison W, Campbell P, Boyle WJ,et al. Osteoprotegerin: a novel secreted protein involved in the regulation of bone density. Cell,1997;89: 309-19. 89309  1997  [PubMed]
 
Ducy P. Cbfa1: a molecular switch in osteoblast biology. Dev Dyn,2000;219: 461-71. 219461  2000  [PubMed]
 
Karsenty G. Role of Cbfa1 in osteoblast differentiation and function. Semin Cell Dev Biol,2000;11: 343-6. 11343  2000  [PubMed]
 
Root AW, Kemp SF, Rundle AC, Dana K,Attie KM. Effect of long-term recombinant growth hormone therapy in children—-the National Cooperative Growth Study, USA, 1985-1994. J Pediatr Endocrinol Metab,1998;11: 403-12. 11403  1998  [PubMed]
 
Faris PM, Ritter MA,Abels RI. The effects of recombinant human erythropoietin on perioperative transfusion requirements in patients having a major orthopaedic operation. The American Erythropoietin Study Group. J Bone Joint Surg Am,1996;78: 62-72. 7862  1996  [PubMed]
 
Gallea S, Lallemand F, Atfi A, Rawadi G, Ramez V, Spinella-Jaegle S, Kawai S, Faucheu C, Huet L, Baron R,Roman-Roman S. Activation of mitogen-activated protein kinase cascades is involved in regulation of bone morphogenetic protein-2-induced osteoblast differentiation in pluripotent C2C12 cells. Bone,2001;28: 491-8. 28491  2001  [PubMed]
 
Clark EA, Golub TR, Lander ES,Hynes RO. Genomic analysis of metastasis reveals an essential role for RhoC. Nature,2000;406: 532-5. 406532  2000  [PubMed]
 
El-Omar EM, Carrington M, Chow WH, McColl KE, Bream JH, Young HA, Herrera J, Lissowska J, Yuan CC, Rothman N, Lanyon G, Martin M, Fraumeni JF Jr,Rabkin CS. Interleukin-1 polymorphisms associated with increased risk of gastric cancer. Nature,2000;404: 398-402. 404398  2000  [PubMed]
 
Collins FS. Shattuck lecture—medical and societal consequences of the Human Genome Project. N Engl J Med,1999;341: 28-37. 34128  1999  [PubMed]
 
Kaji EH,Leiden JM. Gene and stem cell therapies. JAMA,2001;285: 545-50. 285545  2001  [PubMed]
 
Szebik I,Glass KC. Ethical issues of human germ-cell therapy: a preparation for public discussion. Acad Med,2001;76: 32-8. 7632  2001  [PubMed]
 
Shalala D. Protecting research subjects—what must be done. N Engl J Med,2000;343: 808-10. 343808  2000  [PubMed]
 
Friedmann T. Medical ethics. Principles for human gene therapy studies. Science,2000;287: 2163 -5. 2872163  2000  [PubMed]
 
Orlowski JP,Wateska L. The effects of pharmaceutical firm enticements on physician prescribing patterns. There’s no such thing as a free lunch. Chest,1992;102: 270-3. 102270  1992  [PubMed]
 
Wenger NS,Lieberman JR. The orthopaedic surgeon and industry. Ethics and industry incentives. Clin Orthop,2000;378: 39-43. 37839  2000  [PubMed]
 
Swift EM,Yager D. Unnatural selection. Sports Illustrated,2001;94: 86-94. 9486  2001 
 
American Academy of Orthopaedic Surgery, Committee on Ethics 1996-1998. Guide to the ethical practice of orthopaedic surgery. 3rd ed. Rosemont, IL: American Academy of Orthopaedic Surgeons; 1998. p 10-11 
 
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These activities have been planned and implemented in accordance with the Essential Areas and policies of the Accreditation Council for Continuing Medical Education (ACCME) through the joint sponsorship of the American Academy of Orthopaedic Surgeons and The Journal of Bone and Joint Surgery, Inc. The American Academy of Orthopaedic Surgeons is accredited by the ACCME to provide continuing medical education for physicians.
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