Grayson-Jockey Club Research Foundation’s board of directors has announced a slate of 17 research projects which the Foundation will fund for a total of $986,863 in 2015. The list includes ten new projects and seven which are in their second year, as well as Two Research Career Development Awards. The allotment brings the Foundation’s total impact since 1983 to over $22 million to fund 322 projects at 41 universities.
The use of a guinea pig model of Rhodococcus equi would greatly benefit the equine industry. The impact of R. equi pneumonia on the equine industry is large. In the United States, respiratory disease is the third most common cause of disease in foals less than 6 months of age. Foals with R. equi pneumonia can either show clinical signs of disease, or have the lesions without clinical signs (known as subclinical disease), but the most common form of the disease is a chronic pneumonia with an insidious onset that hinder early detection. Often, the lung lesions are well–established and extensive damage has occurred by the time a diagnosis is made. Consequently, a high proportion of foals with R. equi pneumonia die, and foals that do survive the disease are less likely than age–matched cohorts to race.
Treatment and prevention of R. equi pneumonia are difficult: there is no vaccine commercially available, treatment with antimicrobials is lengthy, costly, has potential adverse effects, and is not uniformly successful. Also, there is great need to identify alternative antimicrobials because of evidence of increased resistance. Evaluation of antimicrobials in animals is necessary because drugs that are effective in the laboratory are not always effective in patients. Also, no efficacious vaccine against R. equi pneumonia exists. The equine industry would greatly benefit with the development of both alternative antimicrobials and vaccines that could protect foals against R. equi pneumonia. Thus, there is need to develop a small animal model of R. equi pneumonia, which would allow for evaluation of research questions of clinical importance (such as how the disease develops, and evaluation of new vaccines and antimicrobials) in guinea pigs before they are evaluated in horses or foals. A guinea pig model of R. equi pneumonia would allow for more rapid advances in the field resulting from experiments completed faster than if they were performed in the foal model.
The main hypothesis of this grant is that there are no differences between autologous and allogeneic BMDMSCs in how these cells stimulate the immune system. We will first test this hypothesis by comparing the immunity of these cells in cell culture (in vitro) and attempt to determine the mechanisms by which these cells locally suppress the immune response (Specific Aim 1). In our second Specific Aim (Specific Aim 2) we will compare the immune response to intra–articular injection of each horse when autologous or allogeneic BMDMSCs are administered once and with repeated injection. The completion of this project will answer the important question of whether allogeneic BMDMSCs are a viable alternative to autologous BMDMSCs in the horse. If so, allogeneic cells could be more readily available and less expensive, allowing BMDMSC therapy to be more highly utilized and affordable to owners and trainers.
Regenerative medicine, especially mesenchymal stem cell therapy is a frequently utilized treatment in racehorse and sport horse practice. The lag time of individual cell culture expansion makes this therapy inconvenient and expensive. If allogeneic (non–self) BMDMSCs could be utilized instead of autologous (self) BMDMSCs without these cells being recognized as foreign, this regenerative therapy could be made available as a less expensive “off the shelf” treatment making BMDMSCs available to clients who would have previously foregone the therapy due to expense. Research studies are mounting with evidence of the benefits this therapy provides. When the questions in this proposal are answered, we will have an improved understanding of how these cells avoid immune recognition and potentially have a more available and affordable source of stem cell therapy.
The hypothalamus–pituitary–adrenal gland axis (HPAA) is a complex system that regulates how foals respond to stressful events. Dysfunction of the HPAA results in abnormal concentrations of stress hormones, including steroids from the adrenal gland. These steroids are essential to regulate glucose, electrolytes, blood pressure, as well as organ maturation (lungs, brain, intestine). Some of these steroids (neurosteroids) protect nerve cells against injury and inflammation. We are proposing that in sick newborn foals the equilibrium between these steroid hormones is broken, and depending on its severity it can have deleterious effects on metabolic activities, immune protection, organ function, and likelihood for survival. For example, we suggest that critically ill and premature foals will have increased levels of progesterone (a sex hormone) and low levels of cortisol (a stress hormone).
We will investigate hormones from the brain, pituitary gland, and adrenal gland. We will determine their association with the clinical signs and blood abnormalities of sick foals, as well as whether their levels are linked to clinical improvement or mortality.
We have established collaborations with multiple institutions to investigate the importance of these hormones in the development and progression of diseases affecting newborn foals. We have the expertise to address these questions in a timely manner. The information generated will be relevant to the equine industry as it could lead to the development of novel treatments for foals.
Sepsis is a pathological condition where bacteria can multiply in the blood and tissues, releasing toxins throughout the body of newborn foals. Sepsis is the number one cause of mortality in newborn foals. Many foals that survive sepsis are left with disabilities that impair their future performance. Stress hormones including steroids from the adrenal gland that are essential to control glucose, electrolytes, and blood pressure are often altered. Other steroids are important for organ maturation / function (lungs, brain, intestine). Recent investigations indicate that other steroids (aka neurosteroids) for which minimal information exists in foals protect nerve cells against injury.
We are proposing that in sick newborn foals the equilibrium between these steroid hormones is broken and it can impair their ability to survive.
The goal of this project is to investigate the importance of these hormones in newborn foals with different levels of disease. Hormones will be measured during hospitalization, and their level/balance assessed. The association between hormones, blood abnormalities, and likelihood mortality will be determined.
This study has clinical relevance to the equine industry as a better understanding of equine sepsis will improve treatment. We will generate novel information that will benefit the well–being of horses. Results will be valuable in the implementation of novel therapies to restore the hormonal balance. For example, we may consider using progesterone (a mare hormone) to accelerate brain maturation in dummy foals. In other instances we may want to block the production of other hormones. This project will also be positive for the training of students and veterinarians with interest in equine research. We are confident that we can accomplish the goals of this project in the proposed time–frame.
Insulin resistance is a growing problem among horse populations, and a contributor to such costly and debilitating conditions as obesity and pasture–associated laminitis. While a variety of clinical and field tests are available for the detection of insulin dysregulation, they may be labor intensive, require multiple blood samples, or be imprecise in their ability to assess insulin resistance in individual animals. Identification of a minimally–invasive, single–sample biomarker that allows practitioners to detect and monitor insulin resistance has the potential to advance animal well–being and reduce associated diagnostic and treatment costs to the horse owner. Circulating microRNAs, small molecules involved in gene regulation, have recently been recognized as effective biomarkers to screen for, and even predict, metabolic disease in humans and other species. If this novel technology can be applied to the equine population, it may provide practitioners and horse owners with a new and more convenient tool in the fight against equine metabolic syndrome, and yield important new knowledge about how cellular and physiological processes are regulated in insulin–resistant vs. healthy horses.
Horses with chronic, resistant or repeated bacterial infections are often suspected of harboring biofilm, a carbohydrate–based matrix that coats colonies of bacteria. Biofilms are thought to protect the bacteria from the host immune system and from antimicrobial treatment. Equine–origin bacterial strains are capable of producing a biofilm that is highly resistant to degradation in the laboratory. The purpose of this project is to determine if bacteria isolated from the equine uterus produce a biofilm in the equine uterus and how the host responds to this infection.
The first breakthrough in this area was when it was determined that bacteria in the equine uterus may be associated with a biofilm. The goals of this proposal are to use luciferase producing bacteria to determine the location of bacteria in the uterus for guided biopsy sampling. This will allow the determination of the spatial location and identification of key bacterial sites of colonization and subsequent biofilm formation to determine if the bacteria are on the surface of the endometrium, deep in the glands, or penetrating into the submucosa. Additionally these studies will determine if the bacteria are producing a biofilm as part of the infection, and evaluate the resulting inflammatory response to the bacteria in a biofilm. It is anticipated that this study will provide a better understanding of the role biofilms play in cases of equine infectious endometritis. Future studies, based on results of this pilot study, will focus on evaluating various treatments and how the mare’s immune system is able to help eliminate these infections. Ultimately, these studies will lead to improved therapeutic strategies for management of mares with uterine infections caused by biofilm–producing bacteria.
We have evidence that one of the ways EHV–1 avoids immune responses is by blocking the Type–1 interferon (IFN) response, also known as the IFN–alpha/beta response. The IFN–alpha/beta response is important as a non–specific first line of defense against many virus infections, by making cells more resistant to virus infection. Our preliminary studies show that EHV–1 causes an initial stimulation of IFN–alpha/beta but then blocks its further activity once the virus replication has gone into its late phase. These studies were done using an EHV–1 strain that causes EHM, and we do not know whether the results are applicable to all EHV–1 strains. We propose that one or more factors made by the virus are responsible for blocking the IFN–alpha/beta response, and here we want to find out how that block happens. We also propose that this block happens with the EHV–1 strains that cause EHM but not with the EHV–1 viruses that don’t cause EHM.
Specific aims are:
1. Determine which steps of the IFN–alpha/beta response pathway (sensitization, induction, or amplification) are blocked by EHV–1
2. Determine the relationship between the ability of an EHV–1 strain to cause EHM and its ability to block the IFN–alpha/beta response.
With this knowledge we can potentially design interventions to bolster innate immune responses against EHV–1 and improve control of this disease. This research could eventually lead to treatments for horses infected with EHV–1 that target the viral factors which cause the block in IFN–alpha/beta, with the goal of reducing the severity of disease and the likelihood of EHM.
Our group at the University of Florida treated mares with placentitis using trimethoprim sulfa tablets (antibiotic), pentoxifylline (anti– inflammatory) and Regumate™ (progestin to induce uterine quiescence). Treated mares delivered more live foals (10/12; 83%) when compared to untreated mares (five of five foals dead). Unfortunately, the same treatment protocol used in a clinical setting does not result in this high percentage of live born foals. Theories as to why this treatment protocol fails in some cases include: 1. resistance of bacteria to trimethoprim sulfa, 2. poor ability to diagnose placentitis which causes treatment to be delayed and, 3. uncontrolled inflammation. Research in women has demonstrated that inflammation, with or without bacterial infection, resulted in early delivery of babies. Survival of these babies was poor. Therefore, research efforts have been devoted to early identification and treatment of inflammation in pregnancy. Placentitis in mares closely mimics conditions causing preterm labor in women. It makes perfect sense that our efforts toward improving foal survival from mares with placentitis would involve investigation of drugs that attack inflammation (anti–inflammatory agents).
Our objectives in this work are to examine characteristics of firocoxib, a potent anti–inflammatory drug formulated for use in horses, after it is administered to mares with placentitis. First, we will determine if firocoxib can effectively reduce factors that cause inflammation in placentitis. Second, we will treat mares with placentitis using firocoxib, trimethoprim sulfa and Regumate™ to see if pregnancy is maintained and foal survival improved. We will compare this treatment to a commonly used placentitis treatment of pentoxifylline, trimethoprim sulfa and Regumate™. Our preliminary studies have shown us that firocoxib is found in the fluids of pregnancy (allantoic and amniotic), colostrum and plasma from foals of treated mares. Since firocoxib can penetrate the placenta effectively, we expect that firocoxib will reduce inflammation that occurs after placental infection. Further, we expect that infected mares treated with firocoxib, trimethoprim sulfa and altrenogest will carry pregnancies longer and deliver more live foals than mares administered a more traditional treatment. These objectives fulfill the over–arching goal of our research program which is to improve foal survival after placentitis.
Foals that are delivered prior to the last week of a normal gestation experience high rates of death. Foal death, in turn, exerts a tremendous financial burden on the owner and breeder, both directly, and as reflected in poor produce record for the mare. The single most important cause of abortion and premature delivery in horses is bacterial infection of the placenta (placentitis) which leads to uncontrolled inflammation and premature delivery of a foal. Identification of drugs that are effective for treating mares with placentitis is paramount to improving foal survival. A fundamental step toward making educated therapeutic choices is to determine if specific drugs are performing expected functions, such as resolving inflammation, in mares with placentitis. It is also important to determine if drugs are effective in stalling preterm labor so that the foal has more time to mature, and ideally, survive the infection. Given the high incidence of this condition in equine pregnancy, and the significant financial burden a lost pregnancy imparts, we feel that this work is paramount to the solvency of the equine industry.
Colic is the leading cause of death in horses behind old age according to studies performed by the United States Department of Agriculture. The main reason for death in horses with colic is absorption of bacterial toxins from the gut into the bloodstream. This causes shock, which is often difficult to manage. Treatment has greatly improved over the last 20-30-years, but periodically requires re-evaluation to make sure veterinarians are treating with the best possible mediations. One very important drug used to combat colic is the non-steroidal anti-inflammatory drug (NSAID) flunixin (Banamine®) because it is a painkiller as well as reversing some of the shock-causing effects of bacterial toxins. However, it has side effects, including one that we have recently discovered in which flunixin slows down intestinal healing. This paradoxically increases absorption of bacterial toxins. Interestingly, a new NSAID on the market for horses, firocoxib (Equioxx®), provides more targeted treatment of pain and inflammation while limiting side effects, including delayed intestinal repair. To find out the clinical importance of this potentially critical difference in medications, we want to perform a multi-university clinical trial in which flunixin and firocoxib will be compared in horses going to surgery for colic. We will set it up so that use of firocoxib and flunixin will be given in a random order, and without the veterinarian knowing which drug they are using. In this way, we can remove bias and gather clinical information as well as important indicators of the level of shock that will facilitate decision-making on colic treatment in the future.
Delayed healing and proud flesh formation involving wounds of the horse’s lower limbs is one of the most common problems facing veterinarians and horse owners, and new effective treatments are needed. Our research group is a combination of equine veterinary specialists and biomedical engineers that are working on developing new therapies for wounds or other diseases that are caused by or associated with poor blood flow. Endothelial progenitor cells (EPCs) are cells that form new blood vessels or repair the lining (endothelium) of existing blood vessels. Combining stem or progenitor cells with engineered biomaterials allows the veterinarian to deliver stem cells directly to the area of interest and keep them in that location for a longer period of time. The cells are more likely to have the ability to heal tissues and form blood vessels if they are not immediately carried away. In addition to a potential new treatment for distal limb wounds, the cell/biomaterial combination therapy could have implications for many different diseases and many different cell types.
The investigators of this proposal have recently been funded by GJCRF for two SLL studies using very different models and experimental techniques; however, results from both studies indicate that lamellar hypoxia (meaning decreased tissue oxygen levels, most likely from decreased lamellar blood flow) is the driving force behind lamellar failure in SLL. Because the two laboratories use different approaches (AVE/DWR using a novel microdialysis catheter placed in the lamellar tissue to obtain lamellar interstitial fluid [fluid in the space between the cells which reflects the activities of the cells] for assessment of energy metabolites and blood flow[ by injecting urea into the interstitial space and measuring how long it takes to clear] and JKB using molecular and biochemistry/histologic imaging techniques in lamellar tissue) to detect lamellar cellular dysregulation/ hypoxia, we can now combine these approaches (and our recent results) to fully assess the efficacy of a therapeutic intervention as a prophylactic treatment for the horse at risk of SLL. The results of the van Eps/Richardson study strongly indicate that it is not the excessive weight bearing but is the lack of movement of the supporting limb which results in lamellar hypoxia; they also demonstrated that pharmacologic intervention most commonly used by veterinarians in attempt to increase lamellar blood flow (acepromazine) not only did not improve lamellar metabolism but actually worsened the parameters assessed. Thus, the culmination of these studies indicates that the only effective treatment may be a dynamic shoe which provides the same motion to the digit as walking while allowing the animal to maintain weight on the supporting limb.
Using a custom “V shoe” designed by the Belknap laboratory as a non–painful cause of decreased weight bearing on one limb (and excessive weight bearing on the opposite limb), we are now well situated to test a novel, practical pneumatic shoe device (which was produced by the SoftRide company at our request and to our specifications) on the supporting limb. In the proposed study, we will first perform short term animal protocols (( 6 hours) using a custom stocks present in the van Eps laboratory in order to: 1) determine the ability of different combinations/ cycles of heel and toe elevations to improve lamellar oxygenation/blood flow (assessing lamellar interstitial fluid obtained from the microdialysis catheters inserted in the lamellae in the front fee), and 2) determine the effect of these movements of the supporting limb digit by the pneumatic shoe on the weight bearing on the opposite limb (due to the fact that the opposite/injured limb may only be able to sustain a moderate degree of weight bearing in the clinical case). Once we have established the best combination and frequency of heel/toe elevations which do not cause a dramatic increase in weight bearing on the opposite limb, we will then perform a longer term study in which we will assess (using the microdialysis technique in the van Eps laboratory and biochemistry techniques on tissue samples in the Belknap lab) the efficacy of the pneumatic shoe using the chosen heel/toe elevation protocol (from the first experiment) in horses with the V shoe on the opposite limb for 96 hours.
We expect to determine the efficacy of the use of a pneumatic device to maintain digital movement of the supporting limb in ameliorating the lamellar cellular dysregulation/ hypoxia that occurs with excessive weight bearing on one limb; we feel it is likely that we will establish a protocol using the pneumatic shoe which will be effective in protecting the lamellae from decreased blood flow/hypoxia. In conclusion, the results from the proposed study will not only further our understanding of SLL, but have a great potential of resulting in the rapid establishment of a commercially viable prophylactic treatment for SLL.
Laminitis been voted the number one priority for equine research by the American Association of Equine Practitioners due to both the high incidence of the disease (annual incidence of 2–7% of horses in recent studies), the severe nature of the disease (high incidence of humane destruction or chronic lameness due to crippling nature) and the lack of effective therapies for treating the disease. In one of the largest studies of the incidence of lameness in the U.S. in recent history, a USDA National Animal Health Monitoring Study published in 2000 of approximately 3000 horse farms in 28 states stated that 13% of these farms reported a case of laminitis in a one year period. Supporting limb laminitis occurs in all breeds, and is particularly devastating due to the much higher mortality rate (50%) compared to the other types of laminitis (e.g. endocrinopathic laminitis). Supporting limb laminitis is perhaps the most familiar form of the disease to the racing industry and general public, being the condition that led to the demise of Kentucky Derby winner Barbaro, in 2007 and more recently of Kentucky Derby contender Intense Holiday in 2014. This highlights the fact that, despite great advances in the treatment of even the most catastrophic limb fractures and infections in adult horses, supporting limb laminitis remains the major cause of treatment failure and euthanasia for humane reasons in these cases. Because the horse at risk of supporting limb laminitis is easily detectable in most cases, an effective preventative treatment strategy would be a significant step forward for the welfare of horses and for the horse industry.
From research performed in the past decade it has emerged that there are three distinct major forms of laminitis. One of them is supporting-limb laminitis. Although it has never been proven, supporting limb laminitis is suspected to occur as a result of reduced blood supply to the connection between hoof and bone (the lamellar tissue). It is proposed that horses rely upon regular loading and unloading of the foot in order to move blood (containing nutrients and oxygen) through it. Our research groups (Queensland, U of Penn, and Ohio State), supported by the Grayson-Jockey Club Research Foundation, now have strong preliminary data that supports this theory.
This next step will utilize a protocol that combines real–time monitoring of lamellar tissue blood flow and energy balance with a suitable and humane model of preferential weight bearing on one limb. We seek to confirm the cause of supporting limb laminitis and to test potential therapeutic interventions. It is anticipated that the results of this study will directly guide the design of devices and/or protocols that can be used in the clinical setting to prevent supporting limb laminitis.
An effective preventative strategy would be a significant step forward for the welfare of horses and for the horse industry.
After many decades of frustration with failures of multiple therapies, we now have one therapy, continuous digital hypothermia (CDH), which has proven effective in the laboratory and clinical setting in many horses. We will use advanced biochemistry techniques, combined with two cutting edge research tools including kinome arrays and metabolomics analysis. The results will provide targets for pharmaceutical therapies to either replace or augment CDH, and will also for the first time determine effects of CDH in the normal digit (important to know for any commonly used therapy).
Advancing laminitis therapy will allow more at- risk animals to avoid the crippling consequences of SRL and return to their previous level of function in the equine industry as was witnessed with Paynter.
We presume that these deaths are caused by heatstroke. Our theory is that the overheating problem is similar to anhidrosis (nonsweating), in that the ability to sweat is impaired. We have good evidence for this in the case of erythromycin, based upon studies done in preparation for this proposal. This makes the foal unable to efficiently lose body heat, and on hot days, puts it at risk for heatstroke. We have developed a way to measure precisely the ability of horses to sweat. We plan to use this simple test to show that impaired sweating is the cause of overheating not only in foals given erythromycin but also in some foals treated with azithromycin, clarithromycin, and even the exciting new antibiotic, gamithromycin. Some additional studies will be dedicated to trying to sort out how this class of drugs affects sweating.
If we are right, we hope that these results will serve as a wake-up call for all concerned that foals on erythromycin–like drugs including azithromycin, clarithromycin, and gamithromycin are at risk for heatstroke during and even after treatment and they therefore need to be protected by implementation of common–sense heat control measures.
Data gathered in this study will provide valuable information for the duration of effect of this drug in the horse by measuring up-regulated genes and inflammatory proteins, in both normal and inflamed joints. If, as we have theorized, this drug has a prolonged effect relative to detection times, results from this study may support less frequent administration of intra-articular corticosteroids, which may ultimately prove beneficial to the health of the joint.
Mild and severe injuries to the fetlock are very common. Catastrophic injuries to the fetlock are the greatest cause of death in racehorses, causing about 50% of injury related deaths. Fetlock injuries are due to circumstances that cause the fetlock to extend beyond its normal range of motion (hyperextension). We hypothesize that fetlock hyperextension, and thus related injuries, can be prevented by developing race surfaces that change the way the limb interacts with the surface. Our objective is to determine the characteristics that a race surface should have to prevent fetlock injuries.
It is not economically practical to build the number of race surfaces needed to determine the ideal surface for injury prevention. Thus, a computer modeling and simulation approach is being used because it is a powerful and economical tool for investigation of race surface characteristics on fetlock motion, and thus risk for injury. This approach can be used to assess the effect of surfaces presently installed, as well as those surfaces that do not presently exist. Racehorse fetlock motion will be predicted for a large number of surfaces with different characteristics. Because the predictions are only as good as the computer model, the model will be evaluated for accuracy (validated) by comparing predictions with actual race surface behavior and racehorse fetlock motion that we previously measured simultaneously at two racetracks, one with a dirt surface and one with a synthetic surface. After validation, the model will be used to determine race surface characteristics that prevent fetlock hyperextension. These race surface characteristics can be used as a standard guide.
Unfortunately, routine diagnostic methods (radiographs , ultrasound, MRI, CT) all have shortcomings in evaluating articular cartilage, although MRI is currently the preferred imaging method. However, when contrast agents are injected into the joint prior to CT imaging (CCECT), they have the potential to provide far more information about the health of cartilage. In preliminary experiments we have demonstrated that the amount of CCECT contrast in the cartilage is strongly related to the amount of GAG content within articular cartilage in normal joints. Our hypothesis is that CCECT is a safe and useful method to improve early diagnosis of joint disease. We will test this hypothesis by determining GAG concentrations in normal and diseased articular cartilage and comparing those amounts to CCECT attenuation. We will also test the safety of the contracts agent by evaluating articular cartilage samples for toxicity and lastly, compare the CCECT imaging modality to MRI. It is anticipated that in these experiments we will be able to demonstrate that decreasing GAG content is measurable using contrast CT studies allowing for the early detection of cartilage injury using readily available equipment.
The completion of this project will allow for the critical evaluation of CCECT as a method for the detection of early osteoarthritis in horses and will allow for applications of its use in clinical patients. If this method proves to be effective, it can be a useful tool for monitoring future treatment protocols in horses as well as future cartilage research as it offers a way of ante-mortem in depth evaluation of cartilage health.
The triggering factors for IAD are not known but studies have implicated exposure to barn dust. On the other hand, studies have also supported a role for bacterial infection of the airways. The levels of airborne irritants horses are exposed to in the barn environment can be measured and airway secretions can be sampled in order to classify the type of airway inflammation present in an individual horse. Combining these techniques will allow examination of the relationship between barn exposure and airway inflammation. In the past, investigations of the role of bacteria in IAD have relied upon time-consuming and relatively insensitive culture of mucus. Currently, DNA sequencing is a highly sensitive method that can be used to identify all the bacteria present in mucus, thereby characterizing the entire airway microbe population, or so-called “microbiome.” In addition to bacteria, viruses may cause airway inflammation. Again, DNA sequencing has improved the ability to detect viruses. This study is designed to explore the possibility that the various types of airway inflammation differ in their impact upon racing performance in horses and are due to different causes. Studying these differences is important to develop targeted prevention and treatment strategies.
There are two Career Development Awards offered through the Foundation this year.
The Storm Cat Career Development Award, inaugurated in 2006, is a $15,000 grant designed as an early boost to an individual considering a career in equine research. It has been underwritten annually by Mrs. Lucy Young Hamilton, a Grayson-Jockey Club Research Foundation board member whose family stood the retired champion stallion Storm Cat at Overbrook Farm.
This year the award winner is:
The central hypothesis, based on preliminary data and earlier published reports in other species, is that there are no significant differences in immune profiles between autologous and allogeneic BMDMSCs in vitro and in vivo.
Specific aims include : 1)To compare the immunogenic and the mmunosuppressive properties of autologous to allogeneic bone marrow derived mesenchymal stem cells in vitro and identify a mechanistic explanation for their immunosuppressive properties. The hypothesis is that no significant differences exist between autologous and allogeneic BMDMSCs when compared in vitro and the mechanism of action of immunosuppression may be due to the production of PGE2, indoleamine 2,3-dioxygenase (IDO), TGFß, nitric oxide, or reactive oxygen species. 2) To compare the intra-articular response of autologous versus allogeneic bone marrow derived mesechymal stem cells in vivo and determine if responses change with repeated injections. Our hypothesis is that the joints of horses will not respond significantly differently when autologous or allogeneic BMDMSCs are injected once, or repeatedly.
The Elaine Klein Development Award is a new competitive program intended to promote development of promising investigators by providing a one year salary supplement of $15,000. This program is restricted to one award per year and is named in honor of renowned horsewoman, Elaine Klein. The grant is funded by $15,000 donations by the Klein Family Foundation.
The 2015 award winner is:
The hypothesis is that three-dimensional spheroid culture of equine bone marrow-derived mesenchymal stem cells (eMSCs) will increase production of anti-inflammatory and immunomodulatory mediators and not pro-inflammatory mediators compared to monolayer culture. Futhermore, that spheroid eMSCs will have the following effects in two in vitro models of osteoarthritis: 1) Increase anti-inflammatory/immunomodulatory mediators 2) Increase gene expression of growth factors 3) Reduce the levels of pro-inflammatory mediators 4) Reduce matrix metalloproteinase enzyme (MMP) gene expression and 5) Reduce tissue inhibitor of MMP (TIMP) expression.Specific aims include optimizing the anti-inflammatory properties of eMSCs to treat synovitis and osteoarthritis in horses. This study aims include determining whether: 1)Three-dimensional spheroid culture of eMSCs alters the expression of pro- and anti-inflammatory mediators, MMPs and growth factors compared with monolayer culture. 2) To determine if spheroid or monolayer eMSCs reduce the pro-inflammatory cascade in two in vitro models of synovitis: lipopolysaccharide (LPS)-stimulated synoviocytes and LPS-stimulated cartilage cultures.