Development of quality standards for multi-center, longitudinal magnetic resonance imaging studies in clinical neuroscience

Magnetic resonance imaging (MRI) data is generated by a complex procedure. Many possible sources of error exist which can lead to a worse signal. For example, hidden defective components of a MRI-scanner, changes in the static magnetic field caused by a person simply moving in the MRI scanner room as well as changes in the measurement sequences can negatively affect the signal-to-noise ratio (SNR). A comprehensive, reproducible, quality assurance (QA) procedure is necessary, to ensure reproducible results both from the MRI equipment and the human operator of the equipment. To examine the quality of the MRI data, there are two possibilities. On the one hand, water or gel-filled objects, so-called "phantoms", are regularly measured. Based on this signal, which in the best case should always be stable, the general performance of the MRI scanner can be tested. On the other hand, the actually interesting data, mostly human data, are checked directly for certain signal parameters (e.g., SNR, motion parameters). This thesis consists of two parts. In the first part a study-specific QA-protocol was developed for a large multicenter MRI-study, FOR2107. The aim of FOR2107 is to investigate the causes and course of affective disorders, unipolar depression and bipolar disorders, taking clinical and neurobiological effects into account. The main aspect of FOR2107 is the MRI-measurement of more than 2000 subjects in a longitudinal design (currently repeated measurements after 2 years, further measurements planned after 5 years). To bring MRI-data and disease history together, MRI-data must provide stable results over the course of the study. Ensuring this stability is dealt with in this part of the work. An extensive QA, based on phantom measurements, human data analysis, protocol compliance testing, etc., was set up. In addition to the development of parameters for the characterization of MRI-data, the used QA-protocols were improved during the study. The differences between sites and the impact of these differences on human data analysis were analyzed. The comprehensive quality assurance for the FOR2107 study showed significant differences in MRI-signal (for human and phantom data) between the centers. Occurring problems could easily be recognized in time and be corrected, and must be included for current and future analyses of human data. For the second part of this thesis, a QA-protocol (and the freely available associated software "LAB-QA2GO") has been developed and tested, and can be used for individual studies or to control the quality of an MRI-scanner. This routine was developed because at many sites and in many studies, no explicit QA is performed nevertheless suitable, freely available QA-software for MRI-measurements is available. With LAB-QA2GO, it is possible to set up a QA-protocol for an MRI-scanner or a study without much effort and IT knowledge. Both parts of the thesis deal with the implementation of QA-procedures. High quality data and study results can be achieved only by the usage of appropriate QA-procedures, as presented in this work. Therefore, QA-measures should be implemented at all levels of a project and should be implemented permanently in project and evaluation routines

Effects of Short Term Adiponectin Receptor Agonism on Cardiac Function and Energetics in Diabetic db/db Mice.

Objective Impaired cardiac efficiency is a hallmark of diabetic cardiomyopathy in models of type 2 diabetes. Adiponectin receptor 1 (AdipoR1) deficiency impairs cardiac efficiency in non-diabetic mice, suggesting that hypoadiponectinemia in type 2 diabetes may contribute to impaired cardiac efficiency due to compromised AdipoR1 signaling. Thus, we investigated whether targeting cardiac adiponectin receptors may improve cardiac function and energetics, and attenuate diabetic cardiomyopathy in type 2 diabetic mice. Methods A non-selective adiponectin receptor agonist, AdipoRon, and vehicle were injected intraperitoneally into Eight-week-old db/db or C57BLKS/J mice for 10 days. Cardiac morphology and function were evaluated by echocardiography and working heart perfusions. Results Based on echocardiography, AdipoRon treatment did not alter ejection fraction, left ventricular diameters or left ventricular wall thickness in db/db mice compared to vehicle-treated mice. In isolated working hearts, an impairment in cardiac output and efficiency in db/db mice was not improved by AdipoRon. Mitochondrial respiratory capacity, respiration in the presence of oligomycin, and 4-hydroxynonenal levels were similar among all groups. However, AdipoRon induced a marked shift in the substrate oxidation pattern in db/db mice towards increased reliance on glucose utilization. In parallel, the diabetes-associated increase in serum triglyceride levels in vehicle-treated db/db mice was blunted by AdipoRon treatment, while an increase in myocardial triglycerides in vehicle-treated db/db mice was not altered by AdipoRon treatment. Conclusion AdipoRon treatment shifts myocardial substrate preference towards increased glucose utilization, likely by decreasing fatty acid delivery to the heart, but was not sufficient to improve cardiac output and efficiency in db/db mice

Quality Assurance for functional Magnetic Resonance Imaging

This poster was presented at the OHBM 2016 in Geneva (posternumber: #1696). It describes the introduction of a phantomholder for gel phantom quality assurance to control the stability of the magnetic resonance (MR) signal. Using the phantom holder reduces the variance of the QA metrics and helps the user to detect outlier (or possible MR malfunction) much easier. <br

Development of quality standards for multi-center, longitudinal magnetic resonance imaging studies in clinical neuroscience

Magnetic resonance imaging (MRI) data is generated by a complex procedure. Many possible sources of error exist which can lead to a worse signal. For example, hidden defective components of a MRI-scanner, changes in the static magnetic field caused by a person simply moving in the MRI scanner room as well as changes in the measurement sequences can negatively affect the signal-to-noise ratio (SNR). A comprehensive, reproducible, quality assurance (QA) procedure is necessary, to ensure reproducible results both from the MRI equipment and the human operator of the equipment. To examine the quality of the MRI data, there are two possibilities. On the one hand, water or gel-filled objects, so-called "phantoms", are regularly measured. Based on this signal, which in the best case should always be stable, the general performance of the MRI scanner can be tested. On the other hand, the actually interesting data, mostly human data, are checked directly for certain signal parameters (e.g., SNR, motion parameters). This thesis consists of two parts. In the first part a study-specific QA-protocol was developed for a large multicenter MRI-study, FOR2107. The aim of FOR2107 is to investigate the causes and course of affective disorders, unipolar depression and bipolar disorders, taking clinical and neurobiological effects into account. The main aspect of FOR2107 is the MRI-measurement of more than 2000 subjects in a longitudinal design (currently repeated measurements after 2 years, further measurements planned after 5 years). To bring MRI-data and disease history together, MRI-data must provide stable results over the course of the study. Ensuring this stability is dealt with in this part of the work. An extensive QA, based on phantom measurements, human data analysis, protocol compliance testing, etc., was set up. In addition to the development of parameters for the characterization of MRI-data, the used QA-protocols were improved during the study. The differences between sites and the impact of these differences on human data analysis were analyzed. The comprehensive quality assurance for the FOR2107 study showed significant differences in MRI-signal (for human and phantom data) between the centers. Occurring problems could easily be recognized in time and be corrected, and must be included for current and future analyses of human data. For the second part of this thesis, a QA-protocol (and the freely available associated software "LAB-QA2GO") has been developed and tested, and can be used for individual studies or to control the quality of an MRI-scanner. This routine was developed because at many sites and in many studies, no explicit QA is performed nevertheless suitable, freely available QA-software for MRI-measurements is available. With LAB-QA2GO, it is possible to set up a QA-protocol for an MRI-scanner or a study without much effort and IT knowledge. Both parts of the thesis deal with the implementation of QA-procedures. High quality data and study results can be achieved only by the usage of appropriate QA-procedures, as presented in this work. Therefore, QA-measures should be implemented at all levels of a project and should be implemented permanently in project and evaluation routines

Long-Term Gene Therapy with Thrombospondin 2 Inhibits TGF-b Activation, Inflammation and Angiogenesis in Chronic Allograft Nephropathy

We recently identified Thrombospondin-2 (TSP-2) as a regulator of matrix remodelling and inflammation in experimental kidney disease by using TSP-2 null mice and successfully proved TSP-2 overexpression as a therapeutic concept in a short term glomerulonephritis model in the rat. In this current study, we investigated if long-term TSP-2 overexpression is also capable to ameliorate the progression of chronic kidney disease in the setting of the chronic allograft nephropathy F344-Lewis model in the rat. Two weeks after renal transplantation, two rat thigh muscles were transfected once only with either a TSP-2 overexpressing plasmid (n = 8) or a luciferase-expressing plasmid as control (n = 8). Rats were monitored for renal function, histological changes and gene expression in the graft for up to 30 weeks after transplantation. Unexpectedly, only in the TSP-2 treated group 2 rats died before the end of the experiment and renal function tended to be worsened in the TSP-2 group compared to the luciferase-treated controls. In addition, glomerular sclerosis and tubular interstitial injury as well as cortical fibronectin deposition was significantly increased in the TSP-2 treated kidneys despite reduced TGF-b activation and marked anti-inflammatory (macrophages, T-cells and B-cells) effects in this group. Long-term TSP-2 therapy impaired repair of renal endothelium, as demonstrated by significant higher glomerular and peritubular endothelial rarefaction and reduced endothelial cell proliferation in the transplanted kidneys from TSP-2 treated rats compared to controls. This TSP-2 effect was associated with decreased levels of renal VEGF but not VEGF1 receptor. In conclusion, despit

Long-Term Gene Therapy with Thrombospondin 2 Inhibits TGF-β Activation, Inflammation and Angiogenesis in Chronic Allograft Nephropathy

We recently identified Thrombospondin-2 (TSP-2) as a regulator of matrix remodelling and inflammation in experimental kidney disease by using TSP-2 null mice and successfully proved TSP-2 overexpression as a therapeutic concept in a short term glomerulonephritis model in the rat. In this current study, we investigated if long-term TSP-2 overexpression is also capable to ameliorate the progression of chronic kidney disease in the setting of the chronic allograft nephropathy F344-Lewis model in the rat. Two weeks after renal transplantation, two rat thigh muscles were transfected once only with either a TSP-2 overexpressing plasmid (n = 8) or a luciferase-expressing plasmid as control (n = 8). Rats were monitored for renal function, histological changes and gene expression in the graft for up to 30 weeks after transplantation. Unexpectedly, only in the TSP-2 treated group 2 rats died before the end of the experiment and renal function tended to be worsened in the TSP-2 group compared to the luciferase-treated controls. In addition, glomerular sclerosis and tubular interstitial injury as well as cortical fibronectin deposition was significantly increased in the TSP-2 treated kidneys despite reduced TGF-β activation and marked anti-inflammatory (macrophages, T-cells and B-cells) effects in this group. Long-term TSP-2 therapy impaired repair of renal endothelium, as demonstrated by significant higher glomerular and peritubular endothelial rarefaction and reduced endothelial cell proliferation in the transplanted kidneys from TSP-2 treated rats compared to controls. This TSP-2 effect was associated with decreased levels of renal VEGF but not VEGF1 receptor. In conclusion, despite its anti-inflammatory and TGF-β activation blocking effects, TSP-2 gene therapy did not ameliorate but rather worsened experimental chronic allograft nephropathy most likely via its anti-angiogenic properties on the renal microvasculature

Long-term gene therapy with thrombospondin 2 inhibits TGF-β activation, inflammation and angiogenesis in chronic allograft nephropathy.

We recently identified Thrombospondin-2 (TSP-2) as a regulator of matrix remodelling and inflammation in experimental kidney disease by using TSP-2 null mice and successfully proved TSP-2 overexpression as a therapeutic concept in a short term glomerulonephritis model in the rat. In this current study, we investigated if long-term TSP-2 overexpression is also capable to ameliorate the progression of chronic kidney disease in the setting of the chronic allograft nephropathy F344-Lewis model in the rat. Two weeks after renal transplantation, two rat thigh muscles were transfected once only with either a TSP-2 overexpressing plasmid (n = 8) or a luciferase-expressing plasmid as control (n = 8). Rats were monitored for renal function, histological changes and gene expression in the graft for up to 30 weeks after transplantation. Unexpectedly, only in the TSP-2 treated group 2 rats died before the end of the experiment and renal function tended to be worsened in the TSP-2 group compared to the luciferase-treated controls. In addition, glomerular sclerosis and tubular interstitial injury as well as cortical fibronectin deposition was significantly increased in the TSP-2 treated kidneys despite reduced TGF-β activation and marked anti-inflammatory (macrophages, T-cells and B-cells) effects in this group. Long-term TSP-2 therapy impaired repair of renal endothelium, as demonstrated by significant higher glomerular and peritubular endothelial rarefaction and reduced endothelial cell proliferation in the transplanted kidneys from TSP-2 treated rats compared to controls. This TSP-2 effect was associated with decreased levels of renal VEGF but not VEGF1 receptor. In conclusion, despite its anti-inflammatory and TGF-β activation blocking effects, TSP-2 gene therapy did not ameliorate but rather worsened experimental chronic allograft nephropathy most likely via its anti-angiogenic properties on the renal microvasculature

The mTOR Inhibitor Everolimus Attenuates the Time Course of Chronic Anti-Thy1 Nephritis in the Rat

Background: The antiproliferative immunosuppressant everolimus adversely affects the acute reversible anti-Thy1 nephritis model. We hypothesized that everolimus treatment started after the acute proliferative phase could even be beneficial in the chronic anti-Thy1 nephritis model in the rat. Methods: Chronic anti-Thy1 nephritis was induced by injection of the monoclonal antibody 1-22-3 in 20 male Sprague-Dawley rats 7 days after uninephrectomy. Two weeks after disease induction, rats were randomly treated with either everolimus or vehicle for 14 weeks. Changes in progression of renal disease were investigated by immunohistochemistry and real-time PCR 16 weeks after disease induction. Results: During chronic anti-Thy1 nephritis, the formation of focal segmental glomerulosclerosis lesions, the degree of interstitial fibrosis as well as the increase in proteinuria over 14 weeks was ameliorated by everolimus treatment. Increased glomerular hypertrophy observed in the vehicle-treated rats was completely prevented in the everolimus- treated nephritic rats. Increased glomerular fibronectin mRNA and protein as well as the renal influx of monocytes/ macrophages was significantly reduced in the evero- limus group. Everolimus reduced the pro-angiogenic factor vascular endothelial growth factor (VEGF) and VEGF mRNA in glomeruli, while the transforming growth factor- _ signaling pathway was not affected. Conclusion: ‘Late’ start of everolimus treatment demonstrates beneficial effects on the time course of chronic anti-Thy1 nephritis

Influence of TSP-2 gene therapy on VEGF/VEGF receptor expression and MMP-2 activity.

<p>VEGF and VEGF receptor 1 expression was assessed by semiquantitative scoring of immunohistochemistry. VEGF was reduced in glomeruli (A) as well as in the tubulointerstitial compartment (B) from TSP-2 treated rats. Representative pictures from VEGF stained glomeruli showing pronounced VEGF expression in podocyted as well as mesangial cells in kidneys from control plasmid treated animals (C, brown staining). In contrast, VEGF was only rarely detected in glomeruli from TSP-2 treated rats (D, brown staining). Expression of VEGF receptor 1 was similar in glomeruli (E) as well as cortex (F) of both groups. Glomerular VEGF mRNA expression was comparable in both groups (G). Serum MMP-2 activity at endpoint of the study was detected by zymography (I) and evaluated by densitometry (H). Control (n = 8) vs. TSP-2 treated (A–F:n = 8; G–I:n = 6); *p<0,05.</p