Progress 04/22/09 to 04/21/14
Outputs
Target Audience: The goal of this project is to help veterinarians better diagnose and manage chronic renal disease in dogs. Therefore, the target audience is the veterinary community.During this past year, the target audience was primarily veterinary internal medicine specialists, as that was the subset of veterinarians attending the national meetings at which I presented my talks. The results presented have helped further interest in the subject and emphasize the need for additional research in this area. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? This project has provided numerous training and professional development opportunities. In this most recent reporting period, 2 graduate students have continued to gain valuable experience in benchtop research, design of an experiment, and analysis of results. One has been actively engaged in writing a scientific manuscript to report her findings, and she has presented her results at two additional conferences: the 2013 ACVP/ASVCP Concurrent Annual Meeting in Montreal, Canada, and the 2014 Graduate Student and Postdoc Research Symposium at the Texas A&M College of Veterinary Medicine and Biomedical Sciences. How have the results been disseminated to communities of interest? Since the last report (Sept 30, 2013) results from these studies were presented by a PhD student at 1 international professional meeting (2013 ACVP/ASVCP Concurrent Annual Meeting in Montreal, Canada) and one local meeting (2014 Graduate Student and Postdoc Research Symposium at the Texas A&M College of Veterinary Medicine and Biomedical Sciences). In addition, collection of follow-up information has allowed us to provide additional information regarding kidney disease to veterinarians and owners in several instances. In one situation, we provided the means for an owner (of an Airedale terrier who succumbed to kidney disease) to obtain a diagnosis in her family of dogs caused by a recently identified mutation in Airedales and Soft-coated Wheaten Terriers. She can now resume her breeding program knowing how to avoid perpetuation of these mutations. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Chronic kidney disease (CKD) is a major cause of illness in dogs that can lead to a shortened lifespan and a lower quality of life due to a number of disease complications. We are only starting to fully realize the breadth of kidney diseases present in dogs, and with this increased awareness of its impact on our pet population, we are increasingly recognizing the limitations of current tests of kidney function available in our laboratories. This project has served to increase our experience with and knowledge of potential novel biomarkers of kidney disease in dogs. Biomarkers that are more sensitive and specific for progressive kidney injury are sorely needed in both human and veterinary medicine. As we strive to identify and treat animals earlier in their disease process, having such tests available will be crucial not only for diagnosis but to monitor and adjust therapeutic protocols. The full impact of novel therapies cannot be realized without improved biomarkers of kidney function and damage. In our study, we evaluated several specific urine proteins in dogs with naturally occurring kidney disease and found that these proteins provide a non-invasive tool for earlier and more specific detection of kidney damage in dogs. Importantly, one of these proteins was associated with death due to kidney failure and provided prognostic information beyond conventional tests of kidney function and damage. Ultimately, we will use these results to develop a panel of tests that will be more sensitive and specific for the detection of CKD than currently available tests. Earlier and more accurate identification of disease would allow for earlier treatment that can help slow progression of disease and therefore increase longevity and the quality of life in dogs with CKD. Therefore, this project has the potential to influence every day veterinary clinical practice and to improve the lives of dogs and their owners. Objective 1. To assay selected renal tubular proteins and markers of tubular and glomerular function in serial urine samples from male dogs with X-linked hereditary nephropathy (XLHN) as well as from normal siblings (controls), and correlate these measurements with standard measures of tubular and glomerular function as well as with renal biopsy findings to assess their individual and collective value for detecting onset and gauging progression of chronic kidney disease. This objective was completed prior to the current reported period. 2. To use proteomic techniques to examine the urine protein profiles of these dogs in search of novel biomarkers of early tubular damage and renal disease progression. This objective was completed prior to the current reported period. 3. To further evaluate promising proteins identified by the first 2 objectives using clinical samples submitted to the Texas Veterinary Renal Pathology Service (TVRPS) for diagnostic evaluation. In this past reporting period, we have continued to analyze the data obtained by measuring 5 proteins in the urine and serum of ~200 dogs with spontaneous kidney disease whose samples were collected for diagnostic purposes by clinicians from throughout the United States. In addition, SDS-PAGE analysis of the urine was performed. These samples are paired with a concurrent renal biopsy sample that was extensively evaluated/scored by a veterinary nephropathologist. This comprehensive evaluation serves as the gold standard for kidney damage present in these dogs and allows for the comparison of novel biomarker assays with conventional tests of kidney function and damage. In addition, follow-up clinical information was obtained from ~100 of the 200 dogs evaluated. In addition, the urinary concentration and fractional excretion of one of the novel biomarkers of tubular damage provided a similar estimate of tubular damage severity as did the most widely used, currently available test (serum creatinine (sCr)). However, this novel protein does not share some of the major limitations of sCr and might therefore be a useful adjunct in the diagnosis of significant damage to the tubulointerstitial compartment. Importantly, this protein correlated with death due to kidney failure, and statistical analysis indicated that it was the only novel biomarker (along with biopsy evaluation) to provide prognostic information beyond serum creatinine. Urinary IgG and IgM and fractional excretion of IgG and IgM correlated the most strongly with severity of glomerular damage, and they demonstrated a stronger correlation than urine protein:creatinine ratio (UPC), which is currently the most widely used clinical assay for detecting glomerular damage. These results suggest that measurement of IgG and IgM in the urine of dogs may be helpful in detecting and monitoring glomerular damage. One conventional molecule (serum creatinine (sCr)) and one novel protein (retinal binding protein (RBP)) showed the best correlation with severity of tubulointerstitial damage (TID). Hazard ratio analysis revealed sCr to have the most significant impact on outcome, where every 1-unit increase in sCr increased the hazard ratio risk of death by 51%. We performed further analysis on the effect of active sediment (e.g., pyuria, hematuria) on biomarker results. Some differences were noted in these samples, particularly in dogs with pyuria and/or bacteriuria, although serum creatinine was also much higher in these dogs, which could indicate the higher novel biomarker concentrations are due to worse kidney disease rather than non-renal influences. Regardless, both pyuric and hematuric samples were removed from our final analysis. We have also been evaluating urine sediment for gene expression products that might help detect ongoing glomerular injury. In particular, we aim to detect podocytes, since loss of podocytes is thought to contribute to glomerular damage. We have identified reference genes suitable for qRT-PCR using urine sediment that allow for more accurate comparison of results between samples, and we have finalized our selection of primer/probe sets that detect podocyte-specific genes. We are actively evaluating the urine sediment samples from dogs with CKD to investigate the utility of urinary podocyte mRNA for detecting podocyte loss. By analyzing these samples in the context of the rich corresponding clinical and prognostic data, we have gained a better sense of the usefulness of several proteins that have been implicated as promising biomarkers of renal disease as well as how protein patterns are altered with renal disease. Our results bring into question the acceptance of one of the proteins (NAG) as an indicator of tubular damage. They also suggest that we can gain a better sense of glomerular damage by measuring IgG and IgM as opposed to UPC. The information we have gathered and are continuing to discover is essential in order to understand how proteinuria is manifested in dogs with kidney disease. It is clear that the information gained from this these tests provides new and useful information that cannot be obtained with conventional testing, including a better understanding of kidney disease and urine proteomics. With this knowledge, it is our hope that we will be able to better manage patients with kidney disease using non-invasive and relatively inexpensive testing strategies. While the immediate impact will be in the veterinary field, the pathophysiology of disease is similar to human renal disease. Therefore, there is the potential for impact in human medicine as well. Last, on a personal level, this project has enhanced not only the growth of my first graduate students, but my own professional growth as I am learning to manage a laboratory and guide undergraduate, veterinary, and graduate students in research.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Schneider, S.M., Cianciolo, R, Nabity, M.B., Brown, C.A., Clubb, F., Lees, G.E. Prevalence of immune-complex glomerulonephritides in dogs biopsied for suspected glomerular disease: 501 cases (2007-2012), J Vet Intern Med, 27(s1):S67-S75, 2013.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
J.A. Hokamp, M.B. Nabity, G.E. Lees, R. Cianciolo. Evaluation of NGAL and RBP as tubular biomarkers in naturally occurring canine chronic kidney disease. DOI: 10.1111/vcp.12092
|
Progress 01/01/13 to 09/30/13
Outputs
Target Audience: The goal of this project is to help veterinarians better diagnose and manage chronic renal disease in dogs. Therefore, the target audience is the veterinary community. During this past year, the target audience was primarily veterinary internal medicine specialists, as that was the subset of veterinarians attending the national meetings at which I presented my talks. The results presented have helped further interest in the subject and emphasize the need for additional research in this area. The other major target audience this past year includes veterinary students and residents, since I incorporate some of my findings into my veterinary student and resident teaching. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? This project has provided numerous training and professional development opportunities. In this most recent reporting period, 2 graduate students have gained valuable experience in benchtop research, design of an experiment, and analysis of results. One undergraduate student completed her honor’s thesis, stemming from this project. In addition, one veterinary student gained research experience through this project as a participant in the Veterinary Medical Student Research Training Program, and one of the graduate students gained experience mentoring this veterinary student over the summer. All of these students gained experience with scientific writing and presentation at conferences, including the 2013 Graduate Student and Postdoc Research Symposium at the Texas A&M College of Veterinary Medicine and Biomedical Sciences and the Veterinary Medical Student Research Training Program Conference (which included both local and national presentations). How have the results been disseminated to communities of interest? From Jan 1-Sept 30, 2013, results from these studies were presented at 2 national professional meetings (ACVIM Forum, Seattle, WA; ACVIM Nephrology Course, Las Vegas, NV)). This research also provided the opportunity for a veterinary student and PhD student to present at the conferences listed earlier. In addition, this research is highlighted on the AVMA website as an example of research conducted by AVMA members. This website is available to the public and will help with public understanding of our research activities. What do you plan to do during the next reporting period to accomplish the goals? The major unfinished goal is to obtain more follow-up information from each naturally occurring case in order to better determine the prognostic value of both novel and conventional tests of kidney function and damage. We will then finalize our analysis and complete manuscripts detailing our findings for submission to a veterinary journal.
Impacts
What was accomplished under these goals?
Chronic kidney disease (CKD) is a major cause of illness in dogs. The prevalence of CKD has been reported to be as high as 7% in dogs. However, since many dogs with CKD go unrecognized, particularly in the early stages, the actual percentage of dogs affected may be higher. Kidney disease can lead to a shortened lifespan and a lower quality of life due to a number of disease complications. There are several tests available to aid in the diagnosis of CKD. These include urine tests and blood tests. The urine tests can detect decreased urine concentration and/or increased urine protein, both of which can be indicators of CKD. The blood tests detect substances in the blood that build up due to decreased kidney function, such as creatinine and urea. However, approximately 65-75% of kidney function must be lost to see increases in serum creatinine and decreases in urine concentration. Because of the lack of sensitivity of these tests, CKD is often diagnosed at a late stage, when treatment is less effective. Detection of total urine protein in CKD can be more sensitive and can provide diagnostic and prognostic information, but it can also be non-specific. In our study, we evaluated several specific urine proteins in dogs with naturally occurring kidney disease and found that these proteins provide a non-invasive tool for earlier and more specific detection of kidney damage in dogs. Importantly, one of these proteins was associated with death due to kidney failure and provided prognostic information beyond conventional tests of kidney function and damage. Ultimately, we will use these results to develop a panel of tests that will be more sensitive and specific for the detection of CKD than currently available tests. Earlier and more accurate identification of disease would allow for earlier treatment that can help slow progression of disease and therefore increase longevity and the quality of life in dogs with CKD. Therefore, this project has the potential to influence every day veterinary clinical practice and to improve the lives of dogs and their owners. Objective 1. To assay selected renal tubular proteins and markers of tubular and glomerular function in serial urine samples from male dogs with X-linked hereditary nephropathy (XLHN) as well as from normal siblings (controls), and correlate these measurements with standard measures of tubular and glomerular function as well as with renal biopsy findings to assess their individual and collective value for detecting onset and gauging progression of chronic kidney disease. In addition to urinary biomarkers, we have been evaluating a novel serum biomarker of glomerular filtration rate (GFR): SDMA. Based on results from the males affected with XLHN, SDMA correlates well with GFR. In addition, SDMA is able to identify a decrease in GFR sooner than serum creatinine (sCr) if a standard, single cutoff value is used for each analyte. However, when trending is performed for sCr, it identifies a decrease in GFR at a similar time point as SDMA. These results indicate that trending of sCr is necessary for sensitive detection of decreasing GFR and that SDMA might be a useful adjunct to sCr in identification of renal disease, particularly given the tendency of clinicians to identify azotemia based on a reference interval. This has significant relevance for veterinary practitioners in their assessment of sCr, and the addition of SDMA to a standard biochemistry panel will provide another resource to aid in their diagnosis and monitoring of kidney disease in dogs. 2. To use proteomic techniques to examine the urine protein profiles of these dogs in search of novel biomarkers of early tubular damage and renal disease progression. This objective was completed prior to the current reported period. 3. To further evaluate promising proteins identified by the first 2 objectives using clinical samples submitted to the Texas Veterinary Renal Pathology Service (TVRPS) for diagnostic evaluation. In this past reporting period, we have completed the analysis of 5 proteins in the urine and serum of ~200 dogs with spontaneous kidney disease whose samples were collected for diagnostic purposes by clinicians from throughout the United States. In addition, SDS-PAGE analysis of the urine was performed. These samples are paired with a concurrent renal biopsy sample that was extensively evaluated/scored by a veterinary nephropathologist, the evaluation of which has also been completed. In addition, follow-up clinical information was obtained from ~60 of the 200 dogs evaluated. IgG correlated the most strongly with severity of glomerular damage, and demonstrated a stronger correlation than urine protein:creatinine ratio (UPC), which is currently the most widely used clinical assay for detecting glomerular damage. These results suggest that measurement of IgG in the urine of dogs may be helpful in detecting and monitoring glomerular damage. When glomerular damage was evaluated with electron microscopy, both conventional and novel tests of glomerular damage had increased correlation as compared to evaluation with light microscopy. This suggests that electron microscopy more accurately reflects glomerular function and highlights the benefit of evaluating renal biopsies using both light and electron microscopy. One novel protein (retinal binding protein (RBP)) and one conventional molecule (serum creatinine (sCr)) showed the best correlation with severity of tubulointerstitial damage (TID). sCr correlated most strongly with TID; however, its correlation was similar to that of RBP. This suggests that RBP might be at least as useful as sCr in the detection and monitoring of TID. Given that RBP has different kinetics than sCr and is not impacted by muscle mass of the patient, it could serve as a useful adjunct in evaluating dogs with suspected kidney disease. Although NAG is reported to be a marker of TID, it does not appear to correlate well with biopsy-proven TID in the kidney. Thus, the clinical utility of this test is uncertain, and it may not provide clinically useful information to veterinary practitioners in the monitoring of proteinuric chronic kidney disease. By analyzing these samples in the context of the rich corresponding clinical and prognostic data, we have gained a better sense of the usefulness of several proteins that have been implicated as promising biomarkers of renal disease as well as how protein patterns are altered with renal disease. Our results bring into question the acceptance of one of the proteins (NAG) as an indicator of tubular damage. They also suggest that we can gain a better sense of glomerular damage by measuring IgG as opposed to urine protein creatinine ratio. The information we have gathered and are continuing to discover is essential in order to understand how proteinuria is manifested in dogs with kidney disease. It is clear that the information gained from this these tests provides new and useful information that cannot be obtained with conventional testing, including a better understanding of kidney disease and urine proteomics. With this knowledge, it is our hope that we will be able to better manage patients with kidney disease using non-invasive and relatively inexpensive testing strategies. While the immediate impact will be in the veterinary field, the pathophysiology of disease is similar to human renal disease. Therefore, there is the potential for impact in human medicine as well. Last, on a personal level, this project continues to enhance not only the growth of my first graduate students, but my own professional growth as I am learning to manage a laboratory and guide undergraduate, veterinary, and graduate students in research.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
Nabity, M.B. 2013 Is measuring renal function useful with CKD? Page 1 to 3 in Proceedings, American College of Veterinary Internal Medicine Advanced Education: Nephrology: Something Old, Something New, Las Vegas, NV.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
Nabity, M.B. 2013 Proteinuria Biomarker or treatment target? Page 16 to 18 in Proceedings, American College of Veterinary Internal Medicine Advanced Education: Nephrology: Something Old, Something New, Las Vegas, NV.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
Schneider, S.M., Cianciolo, R, Nabity, M.B., Brown, C.A., Clubb, F., Lees, G.E. Prevalence of immune-complex glomerulonephritides in dogs biopsied for suspected glomerular disease: 501 cases (2007-2012), J Vet Intern Med, 27(s1):S67-S75, 2013
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
M.B. Nabity, G.E. Lees, M.M. Boggess, M. Yerramilli, E. Obare, M. Yerramilli, J. Aguiar, R. Relford. Correlation of symmetric dimethylarginine with glomerular filtration rate in dogs with chronic progressive renal disease, J Vet Intern Med, v27 (3), p733, 2013.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2013
Citation:
M.B. Nabity, G.E. Lees, M.M. Boggess, M. Yerramilli, E. Obare, M. Yerramilli, J. Aguiar, R. Relford. Week-to-week variability of iohexol clearance, serum creatinine, and symmetric dimethylarginine in dogs with stable chronic renal disease, J Vet Intern Med, v27 (3), p734, 2013.
|
Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: Activities: Partial analysis of samples submitted from throughout the United States from dogs with a variety of renal diseases has been completed. This includes urine SDS-PAGE and band analysis, determination of creatinine and total protein, and determination of specific protein concentrations (NAG, IgG, and IgM) on over 200 urine samples. This work is largely being performed by my first PhD graduate student. In addition, we have started to work on evaluation of gene expression products in the urine sediment samples. This includes ensuring the specificity of primer sets for 5 reference genes as well as 3 target genes using conventional PCR and sequencing. We have tested some of the primer/probe sets using qPCR to determine which one(s) will be most appropriate for normalization of our target genes. This portion of the project has enabled a veterinary student and 2 undergraduate students to gain experience with benchtop research. Another portion of the project has involved culturing of urinary cells in dogs with progressive renal disease due to X-linked hereditary nephropathy. This portion of the project has given a clinical pathology resident the opportunity to participate in laboratory research and led to her submission of an internal grant to fund additional studies using these cells. We anticipate this will also serve as the first of experiments for her PhD in my laboratory. Services: Clinical service is provided to clinicians via the urine SDS-PAGE reports, which are sent to the submitting clinicians. Each report includes the results for the urine sample and my interpretation of these results. I am available to consult on the cases when clinicians have questions. In several cases, the urine SDS-PAGE results were helpful in the diagnosis, and in some cases, it has helped the clinician decide whether or not to proceed with a kidney biopsy. We are continuing to evaluate the prognostic value of the results. Products: This research has fostered several collaborations, both national and international. The samples collected for the urine protein electrophoresis will serve as a resource for future investigations for a graduate student. Dissemination: Results from these studies were presented at a national meeting (ACVIM Forum, New Orleans, LA). This research also provided the opportunity for 2 undergraduate students and 1 veterinary student to participate in research, along with 1 PhD student and one potential PhD student. This project has also opened several collaborations with other academic institutions and industry to test and develop better tests for biomarkers of renal disease in serum and urine. In particular, we are collaborating with Drs. Luca Aresu and Silvia Benali from the University of Padova, Italy. Dr. Benali is working on her PhD, and she helped with the analysis of samples during a 3-month visit this past year. PARTICIPANTS: Dr. Mary Nabity (PI) designs and oversees the research experiments associated with this project. This past year, the urine SDS-PAGE analysis has been performed by Eesha Farooqi (an undergraduate student worker). Dr. Jessica Hokamp, a PhD student, has been performing the protein analysis on the clinical samples. Dr. George Lees contributes to the intellectual design of the projects and the data interpretation. Dr. Jan Suchodolski is a collaborator, providing expertise on the experimental designs and analyses. Dr. Rachel Cianciolo is a board-certified anatomic pathologist with expertise in nephropathology. She has been and is continuing to perform extensive light and electron microscopic evaluations of the submitted renal biopsies. Over the past year, this project has provided research training to 1 PhD student, 1 clinical pathology resident/potential PhD student, 1 veterinary student, and 2 undergraduate students. TARGET AUDIENCES: Goals of this project are to help veterinarians better diagnose and manage chronic renal disease in dogs. Therefore, the target audience is the veterinary community, in particular veterinary internists and clinical pathologists. The talk presented this past year was targeted to internal medicine specialists, and the results presented have helped further interest in the subject and emphasize the need for additional research in this area. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Change in knowledge: The samples collected from clinicians throughout the country are a valuable resource as they are paired with a concurrent renal biopsy sample that is being extensively evaluated. By analyzing these samples in the context of the rich corresponding clinical and prognostic data, we have already gained a better sense of the usefulness of several proteins that have been implicated as promising biomarkers of renal disease as well as how protein patterns are altered with renal disease. Our results bring into question the acceptance of one of the proteins (NAG) as an indicator of tubular damage. They also suggest that we can gain a better sense of glomerular damage by measuring IgG as opposed to urine protein creatinine ratio. With regard to protein pattern analysis, we have found that protein bands of certain molecular masses appear to correlate with prognosis in the clinical patients better than renal biopsy analysis. These findings prompted identification of these bands and may lead to additional diagnostic tests for monitoring renal disease. In addition, our analysis of these banding patterns has allowed us to become better equipped at interpreting the results of this technique. On a personal level, this project continues to enhance my own professional growth as I am learning to manage a laboratory and guide undergraduate, veterinary, and graduate students in research. Change in actions: The information we have gathered and are continuing to discover is essential in order to understand how proteinuria is manifested in dogs with kidney disease. To my knowledge, we are the only testing facility in the U.S. that offers urine SDS-PAGE for evaluation of renal disease. My interpretation of the urine SDS-PAGE has continued to be modified as we analyze the results, both in comparison with the renal biopsy findings and with prognostic data. It is clear that the information gained from this test provides new and useful information that cannot be obtained with conventional testing. We are in the process of writing a manuscript regarding this analysis. Change in conditions: The information we are gathering has helped us gain a better understanding of renal disease and urine proteomics. With this knowledge, it is our hope that we will be able to better manage patients with renal disease using non-invasive and relatively inexpensive testing strategies. While the immediate impact will be in the veterinary field, the pathophysiology of disease is similar to human renal disease. Therefore, there is the potential for impact in human medicine as well.
Publications
- Nabity, MB, Lees, GE, Cianciolo, R, Boggess, M, Suchodolski, J, Steiner, J. (2012). Urinary biomarkers of renal disease in dogs with X-linked hereditary nephropathy, J Vet Intern Med, 26(2):282-293.
- Nabity, MB. 2012. New Biomarkers of Kidney Disease: An Update. Proceedings, American College of Veterinary Internal Medicine Annual Forum, New Orleans LA.
|
Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: Activities: Partial analysis of samples submitted from throughout the United States from dogs with a variety of renal diseases has been completed. This includes urine SDS-PAGE and band analysis and determination of specific protein concentrations (NAG, IgG, and IgM) on over 150 urine samples. This portion of the project has provided an opportunity to mentor a PhD student that recently joined our laboratory. This student recently received a highly competitive fellowship, in part to perform research on this project. Services: Clinical service is provided to clinicians via the urine SDS-PAGE reports, which are sent to the submitting clinicians. Each report includes the results for the urine sample and my interpretation of these results. I am available to consult on the cases when clinicians have questions. In several cases, the urine SDS-PAGE results were helpful in the diagnosis, and we are continuing to evaluate the prognostic value of the results. Products: The samples collected for the urine protein electrophoresis will serve as a resource for future investigations for a graduate student. Dissemination: Results from these studies were presented at 1 national meetings (ACVP/ASVCP Concurrent Annual Meeting, Nashville, TN) and 1 international meeting (International Society of Nephrology Forefronts Symposium, Aarhus, Denmark). The specific protein concentration data was presented by a clinical pathology resident (ASVCP meeting). This research also provided the opportunity for 2 undergraduate students to participate in research. Therefore, the results obtained from this project at this point in time have been disseminated in the form of abstracts at national and international meetings. This project has also opened several collaborations with other academic institutions and industry to test and develop better tests for biomarkers of renal disease in serum and urine. In particular, Dr. Nabity spent one week in the laboratory of Dr. Alan Parrish at the University of Missouri collaborating on renal markers of mutual interest. In addition, Dr. Lees hosted Drs. Rachel Cianciolo from North Carolina State University, and Dr. Luca Aresu from the University of Padova, Italy to discuss future collaborations. PARTICIPANTS: PARTICIPANTS: Dr. Mary Nabity (PI) designs and oversees the research experiments associated with this project. This past year, the urine SDS-PAGE analysis has been performed principally by Eesha Farooqi with help from Ryan Brock (both undergraduate student workers). Dr. Jessica Hokamp joined our group as a PhD student, and she has been performing the protein analysis on the clinical samples. Dr. George Lees contributes to the intellectual design of the projects and the data interpretation. Dr. Jan Suchodolski is a collaborator, providing expertise on the experimental designs and analyses. Over the past year, this project has provided research training to 2 undergraduate students and 1 clinical pathology resident/PhD student, with the opportunity for the PhD student to present research at a national meeting for the first time. TARGET AUDIENCES: TARGET AUDIENCES: Goals of this project are to help veterinarians better diagnose and manage chronic renal disease in dogs. Therefore, the target audience is the veterinary community, in particular veterinary internists and clinical pathologists. The talks presented this past year have been targeted to clinical pathologists and nephrologists. The results presented have helped establish interest in the subject and emphasize the need for additional research in this area. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Change in knowledge: The samples collected from clinicians throughout the country are a valuable resource as they are paired with a concurrent renal biopsy sample that has been extensively evaluated. By analyzing these samples in the context of the rich corresponding clinical and prognostic data, we have already gained a better sense of the usefulness of several proteins that have been implicated as promising biomarkers of renal disease as well as how protein patterns are altered with renal disease. Our results bring into question the acceptance of one of the proteins (NAG) as an indicator of tubular damage. With regard to protein pattern analysis, we have found that protein bands of certain molecular masses appear to correlate with prognosis in the clinical patients better than renal biopsy analysis. These findings prompted identification of these bands and may lead to additional diagnostic tests for monitoring renal disease. In addition, our analysis of these banding patterns has allowed us to become better equipped at interpreting the results of this technique. On a personal level, this project continues to enhance my own professional growth. This year, I set up a laboratory to continue work on biomarkers of renal disease, and I have recently taken on a graduate student. The experience I have gained through this project has been crucial in preparing me for these challenges. Change in actions: The information we have gathered and are continuing to discover is essential in order to understand how proteinuria is manifested in dogs with kidney disease. Currently, we are the only testing facility that offers urine SDS-PAGE for evaluation of renal disease. My interpretation of the urine SDS-PAGE has continued to be modified as we analyze the results, both in comparison with the renal biopsy findings and with prognostic data. It is clear that the information gained from this test provides new and useful information that cannot be obtained with conventional testing. We are in the process of writing a manuscript regarding this analysis. Change in conditions: The information we are gathering has helped us gain a better understanding of renal disease and urine proteomics. With this knowledge, it is our hope that we will be able to better manage patients with renal disease using non-invasive and relatively inexpensive testing strategies. While the immediate impact will be in the veterinary field, the pathophysiology of disease is similar to human renal disease. Therefore, there is the potential for impact in human medicine as well.
Publications
- Nabity MB, Lees GE, Cianciolo R, Dangott L, Suchodolski J, Steiner J. 2011. Proteomic analysis of urine from male dogs during early stages of tubulointerstitial injury in a canine model of progressive glomerular disease, Vet Clin Pathol, 40(2):222-36.
- Nabity MB. 2011. Urine protein and microalbumin. In: Nephrology and Urology of Small Animals. Eds J. Bartges & D. Polzin, Chapter 8, 1st edition, pp 58-61; Wiley-Blackwell, Ames, IA.
- Hokamp JA, Nabity MB, Lees GE, Cianciolo R. 2011. Evaluation of urinary biomarkers in naturally occurring canine chronic kidney disease. Vet Clin Pathol, 40(4):601.
|
Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: Activities: The activities completed on the project utilizing samples from colony dogs include completion of the analysis for the discovery component of the project as well as completion of the individual protein determination and analysis. For the project utilizing samples submitted from throughout the United States from dogs with a variety of renal diseases, activities completed include additional analysis of samples (urine SDS-PAGE and band analysis) and gathering of follow-up clinical data to determine prognosis. Services: Clinical service is provided to clinicians via the urine SDS-PAGE reports, which are sent to the submitting clinicians. Each report includes the results for the urine sample and my interpretation of these results. I am available to consult on the cases when clinicians have questions. In several cases, the urine SDS-PAGE results were helpful in the diagnosis, and we have recently found what appears to be prognostic value in the results. Products: The colony dog work from this project comprised the majority of my PhD, which I completed in 2010. The samples collected for the urine protein electrophoresis will serve as a resource for future investigations. Dissemination: Results from these studies were presented at 2 national meetings (ACVIM Annual Forum, Anaheim, CA; ACVP/ASVCP Concurrent Annual Meeting, Baltimore, MD) as well as a regional continuing education seminar (Tarrant County Veterinary Medical Association, Fort Worth, TX). The urine gel electrophoresis results were presented by an undergraduate student worker (ACVIM meeting) or a clinical pathology resident (ASVCP meeting). The clinical pathology resident received a Young Investigator Award in the basic research category. This research also provided the opportunity for another undergraduate student (minority ethnic descent) to participate in research. The research involving the colony dogs was presented by me at each meeting. Therefore, the results obtained from this project at this point in time have been widely disseminated in the form of abstract and continuing education presentations/proceedings at regional and national meetings. This project has also opened several collaborations with other academic institutions and industry to test and develop better tests for biomarkers of renal disease in serum and urine. One of these industries recently arranged a meeting of Key Opinion Leaders in the field of veterinary nephrology, and our group was invited, further providing an opportunity for dissemination of our findings. PARTICIPANTS: Dr. Mary Nabity (PI) designed and carried out all of the experiments pertaining to the dog colony thus far, and this project has enabled her to complete her PhD. She also designs and oversees the urine SDS-PAGE research. The urine SDS-PAGE analysis has been performed by Ryan Brock (an undergraduate student worker) and Dr. Jen Brown (a clinical pathology resident), with contributions made by Matt De La Cruz (undergraduate student involved in an undergraduate research program (UBM)). Dr. George Lees contributes to the intellectual design of the projects and the data interpretation. Dr. Jan Suchodolski was a collaborator, providing expertise on the experimental designs and analyses. Over the past year, this project has provided research training to 2 undergraduate students, 1 clinical pathology resident, and 1 PhD student, with the opportunity to present the research at national meetings. TARGET AUDIENCES: Goals of this project are to help veterinarians better diagnose and manage chronic renal disease in dogs. Therefore, the target audience is the veterinary community, in particular veterinary internists and clinical pathologists. The talks presented this past year have been targeted to general practitioners, internists, and clinical pathologists, thereby influencing the major key audience. The results presented have helped establish interest in the subject and emphasize the need for additional research in this area. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Change in knowledge: The samples from the colony dogs provide a unique opportunity to evaluate proteins over the course of disease in a single patient. In addition, the samples collected from clinicians throughout the country are a valuable resource as they are paired with a concurrent renal biopsy sample that has been extensively evaluated. With both of these sample sets, we have been able to gain a better understanding of how specific proteins as well as protein patterns are altered with renal disease. Using the colony dog samples, one of the specific proteins that appears particularly useful for both early detection and monitoring the progression of renal disease is urinary retinol binding protein. Detection of this protein in the urine may be a useful adjunct to other monitors of renal function that are currently often difficult to interpret, particularly early in the disease process. Other proteins were found to reflect continued renal tubular injury, but not to reflect the severity of decreased renal function. Overall utility of these proteins in a larger subset of dogs with renal disease is warranted and is planned for the next phase of the study. With regard to protein pattern analysis, we have found that protein bands of certain molecular masses appear to correlate with prognosis in the clinical patients better than renal biopsy analysis. These findings have prompted further evaluation of these bands and may lead to additional diagnostic tests for monitoring renal disease. In addition, our analysis of these banding patterns has allowed us to become better equipped at interpreting the results of this technique. On a personal level, this project continues to enhance my own professional growth. I am currently in the process of setting up a laboratory to continue work on biomarkers of renal disease, and the knowledge I have gained through this project has been crucial in preparing me for this endeavor. Change in actions: The information we have gathered and are continuing to discover is essential in order to understand how proteinuria is manifested in dogs with kidney disease. Currently, we are the only testing facility that offers urine SDS-PAGE for evaluation of renal disease. My interpretation of the urine SDS-PAGE has continued to be modified as we analyze the results, both in comparison with the renal biopsy findings and with prognostic data. It is clear that the information gained from this test provides new and useful information that cannot be obtained with conventional testing. We are currently in the process of writing a manuscript regarding this analysis. Change in conditions: The information we are gathering has helped us gain a better understanding of renal disease and urine proteomics. With this knowledge, it is our hope that we will be able to better manage patients with renal disease using non-invasive and relatively inexpensive testing strategies. While the immediate impact will be in the veterinary field, the pathophysiology of disease is similar to human renal disease. Therefore, there is the potential for impact in human medicine as well.
Publications
- No publications reported this period
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: Activities: The activities completed on this project include both proteomic techniques outlined in the project (2-dimensional gel electrophoresis and SELDI), comparing the urine proteome between two early timepoints of renal disease in a research colony of dogs in order to detect early biomarkers of renal disease. In addition, four individual proteins have been analyzed in the urine of these dogs with various methods. Another component of the project is diagnostic proteomic evaluation of urine samples submitted from a variety of dogs with renal disease in order to determine if they have glomerular disease, tubular disease, or both. This portion of the project has provided the opportunity for an undergraduate student worker and a clinical pathology resident to become involved in research and they both plan to present abstracts on the data accumulated later this year. Events: I presented an abstract on the 2-dimensional gel electrophoresis at ACVIM this past summer. I will present an abstract on the SELDI analysis and on the urine SDS-PAGE analysis at ACVIM this summer. Services: With the diagnostic urine SDS-PAGE, I have been consulting with clinicians regarding their results. In one case, the urine SDS-PAGE was instrumental in deciding upon the next diagnostic step to take in a valuable show dog. Products: This work is helping me to fulfill my PhD degree, which I plan to complete later this year. It has also opened several collaborations with industry to help develop better tests for biomarkers of renal disease. PARTICIPANTS: Dr. Mary Nabity (PI) has carried out all of the experiments thus far. She is primarily responsible for deciding the direction of the project. Dr. George Lees contributes to the intellectual design of the project and the data interpretation. Training: Part of this project is providing research experience for both an undergraduate student and a veterinary clinical pathology resident. TARGET AUDIENCES: The target audience is the veterinary community in order to help veterinarians better diagnose and manage chronic renal disease in dogs. I am incorporating some of the data we have collected in an invited talk this summer at a national veterinary conference. PROJECT MODIFICATIONS: None to report.
Impacts
Change in knowledge: The samples we have for evaluation provide a unique opportunity to evaluate proteins over the course of disease in a single patient. Therefore, with this project we are gaining a much better understanding of how these proteins behave and how they can be used for monitoring chronic kidney disease in dogs (or even if they should be used for this purpose in dogs). For example, urinary RBP is detected early in the disease process and continues to increase steadily with disease progression supporting decreased tubular reabsorption of this filtered protein. NAG originates from damaged tubules and its activity tends to fluctuate during disease progression indicating of varying degrees of active tubular injury. This project has also been instrumental in allowing us to do initial test validation, helping us to determine which assays are worth pursuing in the future using urine samples from client-owned dogs. The proteomic analysis portion of this project has been a useful exploration of the urine proteome using some powerful research tools, and our results will help direct our discovery efforts as we intend to further evaluate several of the proteins identified with these techniques. Last, this project continues to enhance my own professional growth. I have gained experience working with various companies and technical support, am learning about the different grant managing agencies at Texas A&M as I manage a research project and budget, and am learning how to effectively budget my time between clinics, teaching, and research. I have also started to gain the experience necessary to organize and run a laboratory, determining what I will need as I set up a laboratory, prioritize experiments, manage budgets, and supervise students. Change in actions: The information we are gathering is going to be instrumental in the way we think about proteinuria in dogs with kidney disease. We are well equipped to be the experts and the instigators in the use of these tests. Currently, we are the only testing facility that offers urine SDS-PAGE for evaluation of renal disease. Change in conditions: With better understanding of renal disease and urine proteomics, we will be able to better manage patients with renal disease. While the immediate impact will be in the veterinary field, the pathophysiology of disease is similar to human renal disease. Therefore, there is the potential for impact in human medicine as well.
Publications
- No publications reported this period
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