Integrating shear wave elastography and estimated glomerular filtration rate to enhance diagnostic strategy for renal fibrosis assessment in chronic kidney disease.

Background: Assessing renal fibrosis non-invasively in patients with chronic kidney disease (CKD) remains a considerable clinical challenge. This study aimed to investigate the diagnostic efficacy of different approaches that combine shear wave elastography (SWE) and estimated glomerular filtration rate (eGFR) in distinguishing between mild fibrosis and moderate-to-severe fibrosis in CKD patients. Methods: In this prospective study, 162 patients underwent renal SWE examinations and renal biopsies. Using SWE, the right renal cortex stiffness was measured, and the corresponding SWE value was recorded. Four diagnostic patterns were used to combine eGFR and SWE value: in isolation, in series, in parallel, and in integration. The receiver operating characteristic (ROC) curve was established, and the area under the ROC curve (AUC) was calculated to quantify diagnostic performance. Sensitivity, specificity, and accuracy were computed. Results: The eGFR demonstrated sensitivity of 68.2% and specificity of 83.8%, whereas the SWE value displayed sensitivity of 84.1% and specificity of 62.2%, yielding a similar AUC (78.2% and 77.8%, respectively). Combining in series improved specificity to 97.3%, superior to other diagnostic patterns (all P values <0.01), but compromised sensitivity to 58.0%. When combined in parallel, the sensitivity increased to 94.3%, exceeding any other strategies (all P values <0.05), but the specificity dropped to 48.7%. The integrated strategy, incorporating eGFR with SWE value via the logistic regression algorithm, exhibited an AUC of 85.8%, outperforming all existing approaches (all P values <0.01), with balanced sensitivity, specificity, and accuracy of 86.4%, 74.3%, and 80.9%, respectively. Conclusions: Using an integrated strategy to combine eGFR and SWE value could improve diagnostic performance in distinguishing between mild renal fibrosis and moderate-to-severe renal fibrosis in patients with CKD, thereby helping clinicians perform a more accurate clinical diagnosis.

Ultrasound-based radiomics analysis in the assessment of renal fibrosis in patients with chronic kidney disease.

PURPOSE: Assessment of renal fibrosis non-invasively in chronic kidney disease (CKD) patients is still a clinical challenge. In this study, we aimed to establish a radiomics model integrating radiomics features derived from ultrasound (US) images with clinical characteristics for the assessment of renal fibrosis severity in CKD patients. METHODS: A total of 160 patients with CKD who underwent kidney biopsy and renal US examination were prospectively enrolled. Patients were classified into the mild or moderate-severe fibrosis group based on pathology results. Radiomics features were extracted from the US images, and a radiomics signature was constructed using the maximum relevance minimum redundancy (mRMR) and least absolute shrinkage and selection operator (LASSO) regression algorithms. Multivariable logistic regression was employed to construct the radiomics model, which incorporated the radiomics signature and the selected clinical variables. The established model was evaluated for discrimination, calibration, and clinical utility in the derivation cohort and internal cross-validation (CV) analysis, respectively. RESULTS: The radiomics signature, consisting of nine identified fibrosis-related features, achieved moderate discriminatory ability with an area under the receiver operating characteristic curve (AUC) of 0.72 (95% confidence interval (CI) 0.64-0.79). By combining the radiomics signature with significant clinical risk factors, the radiomics model showed satisfactory discrimination performance, yielding an AUC of 0.85 (95% CI 0.79-0.91) in the derivation cohort and a mean AUC of 0.84 (95% CI 0.77-0.92) in the internal CV analysis. It also demonstrated fine accuracy via the calibration curve. Furthermore, the decision curve analysis indicated that the model was clinically useful. CONCLUSION: The proposed radiomics model showed favorable performance in determining the individualized risk of moderate-severe renal fibrosis in patients with CKD, which may facilitate more effective clinical decision-making.

Noninvasive Assessment of Renal Fibrosis of Chronic Kidney Disease in Rats by [68Ga]Ga-FAPI-04 Small Animal PET/CT and Biomarkers.

Renal fibrosis is the most common pathological feature and common pathway of progression in chronic kidney disease (CKD). We evaluated [68Ga]Ga-FAPI-04 small animal positron emission tomography/computed tomography (PET/CT) and biomarkers as noninvasive assessments of renal fibrosis (RF) in CKD rats to generate new ideas for clinical diagnosis. A rat model of renal fibrosis was administered adenine by gavage (n = 28), and the control group was given 0.9% NaCl by gavage (n = 20). At different time points (weeks 1, 2, 4, and 6), five rats were randomly selected from the two groups for [68Ga]Ga-FAPI-04 small animal PET/CT imaging. At the same time, the expression of Fibroblast activation protein (FAP) in renal tissue and the expression levels of type III procollagen N-terminal peptide (PIIINP), transforming growth factor (TGF-ß1), Klotho, and sex-determining region Y-box protein 9 (SOX9) in blood and urine were determined. FAP was highly expressed in the renal tissue of rats in the CKD group and expression increased with the progression of renal fibrosis. [68Ga]Ga-FAPI-04 small animal PET/CT examination showed that the uptake of radioactive tracers in the CKD group was higher than that in the control group, and SUVmax (r = 0.9405) and target-to-background ratio (TBR) (r = 0.9392) were positively correlated with renal fibrosis. The serum levels of PIIINP, TGF-ß1, and SOX9 in CKD rats were significantly higher than those in the control group and were positively correlated with RF (r = 0.8234, r = 0.7733, and r = 0.7135, respectively) and SUVmax (r = 0.8412, r = 0.7763, and r = 0.6814, respectively). Compared with the control group, the level of serum Klotho decreased and was negatively correlated with RF (r = -0.6925) and SUVmax (r = -0.6322). Compared with the control group, the levels of PIIINP and TGF-ß1 in urine were positively correlated with RF (r = 0.8127 and r = 0.8077, respectively) and SUVmax (r = 0.8400 and r = 0.8177, respectively). Urine Klotho decreased compared with the control group and was negatively correlated with RF (r = -0.5919) and SUVmax (r = -0.5995). The change in urine SOX9 was not statistically significant. In conclusion, compared with renal biopsy, [68Ga]Ga-FAPI-04 small animal PET/CT shows renal fibrosis quickly and noninvasively. PIIINP, TGF-ß1, and Klotho in serum and urine may be used as biomarkers of RF, and serum SOX9 is expected to become a new diagnostic biomarker of RF.

Assessment of Renal Fibrosis in Patients With Chronic Kidney Disease Using Shear Wave Elastography and Clinical Features: A Random Forest Approach.

OBJECTIVE: Renal fibrosis is the common pathological hallmark of chronic kidney disease (CKD) progression. In this study, a random forest (RF) classifier based on 2-D shear wave elastography (SWE) and clinical features for the differential severity of renal fibrosis in patients with CKD is proposed. METHODS: A total of 162 patients diagnosed with CKD who underwent 2-D SWE and renal biopsy were prospectively enrolled from April 2019 to December 2021 and then randomized into training (n = 114) and validation (n = 48) cohorts at a ratio of 7:3. The least absolute shrinkage and selection operator (LASSO) regression and recursive feature elimination for support vector machines (SVM-RFE) algorithm were employed to select renal fibrosis-related features from clinical information and elastosonographic findings. An RF model was subsequently constructed using the aforementioned informative parameters in the training cohort and evaluated in terms of discrimination, calibration and clinical utility in both cohorts. RESULTS: The LASSO and SVM-RFE analyses revealed that age, sex, blood urea nitrogen, renal resistive index, hypertension and the 2D-SWE value were independent risk variables associated with renal fibrosis severity. The established RF model incorporating these six variables exhibited fine discrimination in both the derivation (area under the curve [AUC]: 0.84, 95% confidence interval [CI]: 0.76-0.91) and validation (AUC: 0.88, 95% CI: 0.77-0.98) cohorts. Moreover, the calibration curve revealed satisfactory predictive accuracy, and the decision curve analysis revealed a significant clinical net benefit. CONCLUSION: The developed RF model, via a combination of the 2-D SWE value and clinical information, indicated satisfactory diagnostic performance and clinical practicality toward differentiating moderate-severe from mild renal fibrosis, which may provide critical insight into risk stratification for patients with CKD.

Development and Deployment of a Novel Diagnostic Tool Based on Conventional Ultrasound for Fibrosis Assessment in Chronic Kidney Disease.

RATIONALE AND OBJECTIVES: Accurate identification of risk information about fibrosis severity is crucial for clinical decision-making and clinical management of patients with chronic kidney disease (CKD). This study aimed to develop an ultrasound (US)-derived computer-aided diagnosis tool for identifying CKD patients at high risk of developing moderate-severe renal fibrosis, in order to optimize treatment regimens and follow-up strategies. MATERIALS AND METHODS: A total of 162 CKD patients undergoing renal biopsies and US examinations were prospectively enrolled and randomly divided into training (n = 114) and validation (n = 48) cohorts. A multivariate logistic regression approach was employed to develop the diagnostic tool named S-CKD for differentiating moderate-severe renal fibrosis from mild one in the training cohort by integrating the significant variables, which were screened out from demographic characteristics and conventional US features via the least absolute shrinkage and selection operator regression algorithm. The S-CKD was then deployed as both an online web-based and an offline document-based, easy-to-use auxiliary device. In both the training and validation cohorts, the S-CKD's diagnostic performance was evaluated through discrimination and calibration. The clinical benefit of using S-CKD was revealed by decision curve analysis (DCA) and clinical impact curves. RESULTS: The proposed S-CKD achieved an area under the receiver operating characteristic curve of 0.84 (95% confidence interval (CI): 0.77-0.91) and 0.81 (95% CI: 0.68-0.94) in the training and validation cohorts, respectively, indicating satisfactory diagnosis performance. Results of the calibration curves showed that S-CKD has excellent predictive accuracy (Hosmer-Lemeshow test: training cohort, p = 0.497; validation cohort, p = 0.205). The DCA and clinical impact curves exhibited a high clinical application value of the S-CKD at a wide range of risk probabilities. CONCLUSION: The S-CKD tool developed in this study is capable of discriminating between mild and moderate-severe renal fibrosis in patients with CKD and achieving promising clinical benefits, which may aid clinicians in personalizing medical decision-making and follow-up arrangement.

A methodological study of 2D shear wave elastography for noninvasive quantitative assessment of renal fibrosis in patients with chronic kidney disease.

PURPOSE: To determine the optimal measurement method of 2D shear wave elastography (2D-SWE) for noninvasive quantitative assessment of renal fibrosis in chronic kidney disease (CKD) patients. METHODS: A total of 190 CKD patients were enrolled for 2D-SWE of right kidney. The success rates, coefficients of variation (CV), and pathological correlation of different measurement sites, body positions, and depths were compared. RESULTS: (1) Measurement sites: Success rate in the middle part (100%) was higher than that in the lower pole (97.3%, P > 0.05). CV in the middle part (10.2%) was lower than that in the lower pole (16.4%, P < 0.05). Pathological correlation of the middle part (r = - 0.452, P < 0.05) was higher than that of the lower pole (r = 0.097, P > 0.05). (2) Body positions: Success rate in left lateral decubitus position (100%) was higher than that in supine (99.4%, P > 0.05) and prone position (99.4%, P > 0.05). CV was lowest (11.9%) and pathological correlation was highest (r = -0.256, P < 0.05) in prone position. (3) Measurement depths: Success rate at depth < 4 cm (100%) was higher than that at depth ≥ 4 cm (98.8%, P > 0.05). CV at depth < 4 cm (11.1%) was lower than that at depth ≥ 4 cm (14.4%, P < 0.05). Pathological correlation at depth < 4 cm (r = - 0.303, P < 0.05) was higher than that at depth ≥ 4 cm (r = - 0.156, P > 0.05). CONCLUSION: The optimal measurement method of 2D-SWE for renal fibrosis assessment was prone position, renal middle part, and measurement depth < 4 cm.

A nomogram based on shear wave elastography for assessment of renal fibrosis in patients with chronic kidney disease.

BACKGROUND: Non-invasive evaluation of renal fibrosis is still challenging. This study aimed to establish a nomogram based on shear wave elastography (SWE) and clinical features for the assessment of the severity of renal fibrosis in patients with chronic kidney disease (CKD). METHODS: One hundred and sixty-two patients with CKD who underwent kidney biopsy and SWE examination were prospectively enrolled between April 2019 and December 2021. Patients were classified into mildly or moderately-severely impaired group based on pathology results. All patients were randomly divided into a training (n = 113) or validation cohort (n = 49). Least absolute shrinkage and selection operator (LASSO) algorithm was used for data dimensionality reduction and feature selection. Then, a diagnostic nomogram incorporating the selected features was constructed using multivariable logistic regression analysis. Nomogram performance was evaluated for discrimination, calibration, and clinical utility in training and validation cohorts. RESULTS: The established SWE nomogram, which integrated SWE value, hypertension, and estimated glomerular filtration rate, showed fine calibration and discrimination in both training (area under the receiver operator characteristic curve (AUC) = 0.94; 95% confidence interval (CI) 0.89-0.98) and validation cohorts (AUC = 0.84; 95% CI 0.71-0.96). Significant improvement in net reclassification and integrated discrimination indicated that the SWE value is a valuable biomarker to assess moderate-severe renal impairment. Furthermore, decision curve analysis revealed that the SWE nomogram has clinical value. CONCLUSION: The proposed SWE nomogram showed favorable performance in determining individualized risk of moderate-severe renal pathological impairment in patients with CKD, which will help to facilitate clinical decision-making.

Noninvasive assessment of renal function and fibrosis in CKD patients using histogram analysis based on diffusion kurtosis imaging.

PURPOSE: To investigate the potential of histogram analysis based on diffusion kurtosis imaging (DKI) in evaluating renal function and fibrosis associated with chronic kidney disease (CKD). MATERIALS AND METHODS: Thirty-six CKD patients were enrolled, and DKI was performed in all patients before the renal biopsy. The histogram parameters of diffusivity (D) and kurtosis (K) were obtained using FireVoxel. The histogram parameters between the stable [estimated glomerular filtration rate (eGFR) ≥ 60 ml/min/1.73 m2] and impaired (eGFR < 60 ml/min/1.73 m2) eGFR group were compared. Besides, patients were classified into mild, moderate, and severe fibrosis group using a semi-quantitative standard. The correlations of histogram parameters with eGFR and fibrosis scores were investigated and the diagnostic performances of histogram parameters in assessing renal dysfunction and fibrosis were analyzed. The added value of combination of most significant parameter with 24 h urinary protein (24 h-UPRO) in evaluating fibrosis was also explored. RESULTS: Seven D histogram parameters in cortex (mean, median, 10th, 25th, 75th, 90th percentiles and entropy), two D histogram parameters in medulla (75th, 90th percentiles), seven K histogram parameters in cortex (mean, min, median, 10th, 25th, 75th, 90th percentiles) and three K histogram parameters in medulla (mean, median, 25th percentile) were significantly different between the two groups. The Dmean of cortex was the most relevant parameter to eGFR (r = 0.648, P < 0.001) and had the largest area under the curve (AUC) for differentiating the stable from impaired eGFR group [AUC = 0.889; 95% confidence interval (CI) 0.728-0.970]. The K90th of cortex presented the strongest correlation with fibrosis scores (r = 0.575, P < 0.001) and achieved the largest AUC for distinguishing the mild from moderate to severe fibrosis group (AUC = 0.849, 95% CI 0.706-0.993). Combining the K90th in cortex with 24 h-UPRO gained statistically higher AUC value (AUC = 0.880, 95% CI 0.763-0.996). CONCLUSION: Histogram analysis based on DKI is practicable for the noninvasive assessment of renal function and fibrosis in CKD patients.

Assessment of Fasudil on Contrast-Associated Acute Kidney Injury Using Multiparametric Renal MRI.

Aims: To evaluate the utility of fasudil in a rat model of contrast-associated acute kidney injury (CA-AKI) and explore its underlying mechanism through multiparametric renal magnetic resonance imaging (mpMRI). Methods: Experimental rats (n = 72) were grouped as follows: controls (n = 24), CA-AKI (n = 24), or CA-AKI + Fasudil (n = 24). All animals underwent two mpMRI studies (arterial spin labeling, T1 and T2 mapping) at baseline and post iopromide/fasudil injection (Days 1, 3, 7, and 13 respectively). Relative change in renal blood flow (ΔRBF), T1 (ΔT1) and T2 (ΔT2) values were assessed at specified time points. Serum levels of cystatin C (CysC) and interleukin-1ß (IL-1ß), and urinary neutrophil gelatinase-associated lipocalin (NGAL) concentrations were tested as laboratory biomarkers, in addition to examining renal histology and expression levels of various proteins (Rho-kinase [ROCK], α-smooth muscle actin [α-SMA]), hypoxia-inducible factor-1α (HIF-1α), and transforming growth factor-ß1 (TGF-ß1) that regulate renal fibrosis and hypoxia. Results: Compared with the control group, serum levels of CysC and IL-1ß, and urinary NGAL concentrations were clearly increased from Day 1 to Day 13 in the CA-AKI group (all p < 0.05). There were significant reductions in ΔT2 values on Days 1 and 3, and ΔT1 reductions were significantly more pronounced at all time points (Days 1-13) in the CA-AKI + Fasudil group (vs. CA-AKI) (all p < 0.05). Fasudil treatment lowered expression levels of ROCK-1, and p-MYPT1/MYPT1 proteins induced by iopromide, decreasing TGF-ß1 expression and suppressing both extracellular matrix accumulation and α-SMA expression relative to untreated status (all p < 0.05). Fasudil also enhanced PHD2 transcription and inhibition of HIF-1α expression after CA-AKI. Conclusions: In the context of CA-AKI, fasudil appears to reduce renal hypoxia, fibrosis, and dysfunction by activating (Rho/ROCK) or inhibiting (TGF-ß1, HIF-1α) certain signaling pathways and reducing α-SMA expression. Multiparametric MRI may be a viable noninvasive tool for monitoring CA-AKI pathophysiology during fasudil therapy.

Artificial Intelligence Assessment of Renal Scarring (AIRS Study).

Background: The goal of the Artificial Intelligence in Renal Scarring (AIRS) study is to develop machine learning tools for noninvasive quantification of kidney fibrosis from imaging scans. Methods: We conducted a retrospective analysis of patients who had one or more abdominal computed tomography (CT) scans within 6 months of a kidney biopsy. The final cohort encompassed 152 CT scans from 92 patients, which included images of 300 native kidneys and 76 transplant kidneys. Two different convolutional neural networks (slice-level and voxel-level classifiers) were tested to differentiate severe versus mild/moderate kidney fibrosis (≥50% versus <50%). Interstitial fibrosis and tubular atrophy scores from kidney biopsy reports were used as ground-truth. Results: The two machine learning models demonstrated similar positive predictive value (0.886 versus 0.935) and accuracy (0.831 versus 0.879). Conclusions: In summary, machine learning algorithms are a promising noninvasive diagnostic tool to quantify kidney fibrosis from CT scans. The clinical utility of these prediction tools, in terms of avoiding renal biopsy and associated bleeding risks in patients with severe fibrosis, remains to be validated in prospective clinical trials.

Assessment of Fibrinogen-like 2 (FGL2) in Human Chronic Kidney Disease through Transcriptomics Data Analysis.

Fibrinogen-like 2 (FGL2) was recently found to be associated with fibrosis in a mouse model of kidney damage and was proposed as a potential therapeutic target in chronic kidney disease (CKD). We assessed the association of renal FGL2 mRNA expression with the disease outcome in two independent CKD cohorts (NEPTUNE and Innsbruck CKD cohort) using Kaplan Meier survival analysis. The regulation of FGL2 in kidney biopsies of CKD patients as compared to healthy controls was further assessed in 13 human CKD transcriptomics datasets. The FGL2 protein expression in human renal tissue sections was determined via immunohistochemistry. The regulators of FGL2 mRNA expression in renal tissue were identified in the co-expression and upstream regulator analysis of FGL2-positive renal cells via the use of single-cell RNA sequencing data from the kidney precision medicine project (KPMP). Higher renal FGL2 mRNA expression was positively associated with kidney fibrosis and negatively associated with eGFR. Renal FGL2 mRNA expression was upregulated in CKD as compared with healthy controls and associated with CKD progression in the Innsbruck CKD cohort (p-value = 0.0036) and NEPTUNE cohort (p-value = 0.0048). The highest abundance of FGL2 protein in renal tissue was detected in the thick ascending limb of the loop of Henle and macula densa, proximal tubular cells, as well as in glomerular endothelial cells. The upstream regulator analysis identified TNF, IL1B, IFNG, NFKB1, and SP1 as factors potentially inducing FGL2-co-expressed genes, whereas factors counterbalancing FGL2-co-expressed genes included GLI1, HNF1B, or PPARGC1A. In conclusion, renal FGL2 mRNA expression is elevated in human CKD, and higher FGL2 levels are associated with fibrosis and worse outcomes.

Metabolomic assessment of mechanisms underlying anti-renal fibrosis properties of petroleum ether extract from Amygdalus mongolica.

CONTEXT: The petroleum ether extract (PET) of Amygdalus mongolica (Maxim.) Ricker (Rosaceae) has an ameliorative effect on renal fibrosis (RF). OBJECTIVE: To evaluate the antifibrotic effects of A. mongolica seeds PET on RF by serum metabolomics, biochemical and histopathological analyses. MATERIALS AND METHODS: Sixty male Sprague-Dawley rats were randomly divided into the sham-operated, RF model, benazepril hydrochloride-treated model (1.5 mg/kg) and PET-treated (1.75, 1.25, 0.75 g/kg) groups, and the respective drugs were administered intragastrically for 21 days. Biochemical indicators including BUN, Scr, HYP, SOD, and MDA were measured. Haematoxylin and eosin and Masson staining were used for histological examination. The serum metabolomic profiles were determined by UPLC-Q-TOF/MS and metabolism network analysis. Acute toxicity test was performed to validate biosafety. RESULTS: The PET LD50 was >23.9 g/kg in rats. PET significantly alleviated fibrosis by reducing the levels of Scr (from 34.02 to 32.02), HYP (from 403.67 to 303.17) and MDA (from 1.84 to 1.73), and increasing that of SOD (from 256.42 to 271.85). Metabolomic profiling identified 10 potential biomarkers, of which three key markers were significantly associated with RF-related pathways including phenylalanine, tyrosine and tryptophan biosynthesis, amino sugar and nucleotide sugar metabolism and tyrosine metabolism. In addition, three key biomarkers were restored to baseline levels following PET treatment, with the medium dose showing optimal effect. CONCLUSIONS: These findings revealed the mechanism of A. mongolica PET antifibrotic effects for RF rats on metabolic activity and provided the experimental basis for the clinical application.

Diffusion kurtosis imaging for the assessment of renal fibrosis of chronic kidney disease: A preliminary study.

PURPOSE: To investigate the potential of diffusion kurtosis imaging (DKI) for the assessment of renal fibrosis in chronic kidney disease (CKD), using histopathology as the reference standard. METHODS: Eighty-nine CKD patients and twenty healthy volunteers were recruited in this study. DKI was performed in all participants and all CKD patients received renal biopsy. The values of mean diffusivity (MD) and mean kurtosis (MK) in the renal cortex and medulla were compared between CKD patients and healthy volunteers. The Spearman correlation coefficient was calculated to assess the relationship between MD, MK values and the estimated glomerular filtration rate (eGFR), serum creatinine (SCr), 24 h urinary protein (24 h-UPRO), histopathological fibrosis score. RESULTS: The medullary MD values were significantly lower than cortex, while the cortical MK values were significantly lower than medulla for all participants. Renal parenchymal MD values were significantly lower in the CKD patients than healthy controls, whereas MK values were significantly higher in the CKD patients than healthy controls. In the CKD patients, the significantly negative correlation was observed between the renal parenchymal MD values and the 24 h-UPRO, SCr, histopathological fibrosis score, as well as between the renal parenchymal MK values and the eGFR, while the significantly positive correlation was found between the renal parenchymal MD values and the eGFR, as well as between the renal parenchymal MK values and the 24 h-UPRO, SCr, histopathological fibrosis score. CONCLUSION: DKI shows great potential in the noninvasive assessment of renal fibrosis in CKD.

Assessment of renal fibrosis in a rat model of unilateral ureteral obstruction with diffusion kurtosis imaging: Comparison with α-SMA expression and 18F-FDG PET.

PURPOSE: To investigate the utility of diffusion kurtosis imaging (DKI) MRI for evaluation of renal fibrosis in rats with unilateral ureteral obstruction (UUO). METHODS: Twenty-five rats had UUO, and ten rats were subjected to sham operation as control. DKI was performed on a 3.0 T MRI scanner on days 1, 3, 5, and 7 after ligation. All rats then underwent 18F-FDG dynamic PET to evaluate unilateral renal function, followed by histological analysis to examine α-smooth muscle actin (α-SMA) expression. DKI metrics were assessed among the time points and between two sides, and compared with maximum standardized uptake value (SUVmax), serum levels of creatinine and urea, and fibrosis marker α-SMA. RESULTS: Mean kurtosis (MK) on day 7, axial kurtosis (Ka) on days 3 and 7, mean diffusivity (MD) on days 1, 3, 5, and 7, and fractional anisotropy (FA) on days 3, 5, and 7 of cortex and medulla between the UUO and contralateral sides were significantly different (all p < 0.05). Over the course of UUO progression, there were significant changes in Ka, MD and FA of medulla (all p < 0.05). FA of medulla was positively correlated with SUVmax (r = 0.641, p < 0.001), and MD of cortex was negatively correlated with urea (r = -0.534, p = 0.001). MD of cortex was negatively correlated with α-SMA on UUO sides (r = -0.710, p < 0.001). CONCLUSIONS: DKI shows the potential for noninvasive assessment of renal fibrosis and unilateral renal function induced by UUO.

Intravoxel incoherent motion MRI-derived parameters and T2* relaxation time for noninvasive assessment of renal fibrosis: An experimental study in a rabbit model of unilateral ureter obstruction.

PURPOSE: To evaluate the association of apparent diffusion coefficient (ADC), intravoxel incoherent motion (IVIM) MRI-derived parameters, and T2* relaxation time with histopathological changes observed during renal fibrogenesis in a rabbit model of unilateral ureter obstruction (UUO). METHODS: Twenty New Zealand White rabbits underwent baseline MRI followed by surgery (sham or UUO) and then follow-up MRI at postoperative day (POD) 0, 3, 7, and 14. Hematoxylin and eosin and Masson's trichrome staining was performed to evaluate cell density and area of fibrosis. Spearman rank correlation and Pearson correlation tests and one-way analysis of variance were used for statistical analyses. RESULTS: There was a continuous increase in the area of fibrosis and cell density: rho = 0.900 (95% confidence interval [CI] = 0.760, 0.960; p < 0.0001) and 0.904 (95% CI = 0.769, 0.962; p < 0.0001), respectively. There was a tendency for all MRI variables to decrease at POD 3 and partly recover at POD 7. ADC, D, f, and T2* relaxation time showed significant correlation with area of fibrosis and cell density (r = -0.5177 and -0.6962, -0.5395 and -0.7851, -0.7168 and -0.7902, and -0.6808 and -0.7212, respectively; p = 0.0052-0.0481) while D* did not (p = 0.1997 and 0.7853, respectively). CONCLUSIONS: ADC, IVIM MRI-derived parameters, and T2* relaxation time were significantly associated with the area of fibrosis and cell density during renal fibrogenesis in a rabbit model of UUO. After validation in future studies, MRI may have potential for noninvasive assessment modality of renal fibrosis.

Assessment of renal fibrosis in murine diabetic nephropathy using quantitative magnetization transfer MRI.

PURPOSE: Renal fibrosis is a hallmark of progressive renal disease; however, current clinical tests are insufficient for assessing renal fibrosis. Here we evaluated the utility of quantitative magnetization transfer MRI in detecting renal fibrosis in a murine model of progressive diabetic nephropathy (DN). METHODS: The db/db eNOS-/- mice, a well-recognized model of progressive DN, and normal wild-type mice were imaged at 7T. The quantitative magnetization transfer data were collected in coronal plane using a 2D magnetization transfer prepared spoiled gradient echo sequence with a Gaussian-shaped presaturation pulse. Parameters were derived using a two-pool fitting model. A normal range of cortical pool size ratio (PSR) was defined as Mean±2SD of wild-type kidneys (N = 20). The cortical regions whose PSR values exceeded this threshold (threshold PSR) were assessed. The correlations between the PSR-based and histological (collagen IV or picrosirius red stain) fibrosis measurements were evaluated. RESULTS: Compared with wild-type mice, moderate increases in mean PSR values and scattered clusters of high PSR region were observed in cortex of DN mouse kidneys. Abnormally high PSR regions (% area) that were detected by the threshold PSR were significantly increased in renal cortexes of DN mice. These regions progressively increased on aging and highly correlated with histological fibrosis measures, while the mean PSR values correlated much less. CONCLUSION: Renal fibrosis in DN can be assessed by the quantitative magnetization transfer MRI and threshold analysis. This technique may be used as a novel imaging biomarker for DN and other renal diseases.

Intravoxel incoherent motion diffusion-weighted imaging for the assessment of renal fibrosis of chronic kidney disease: A preliminary study.

PURPOSE: To investigate the potential of Intravoxel incoherent motion diffusion-weighted imaging(IVIM-DWI) for the assessment of renal fibrosis in chronic kidney disease (CKD), using histopathology as a reference standard. METHODS: Eighty-five CKD patients and twenty healthy volunteers were recruited in this study. IVIM-DWI was performed in all of the participants, and all of the CKD patients underwent renal biopsy. The mean values of the true diffusion coefficient (D), pseudo diffusion coefficient (D*) and perfusion fraction (f) in the renal cortex and medulla were compared between the CKD patients and healthy volunteers. The Spearman correlation coefficient was calculated to assess the relationship between the D, D*,f values and the estimated glomerular filtration rate (eGFR), serum creatinine level (SCr), 24h urinary protein level (24h-UPRO), histopathological fibrosis scores. RESULTS: The D, D* and f values were significantly lower in medulla than in the cortex for all of the participants. All of the IVIM parameters were significantly lower in the CKD patients than in the healthy controls. In the CKD patients, a significant negative correlation was found between the renal parenchymal D, D*,f values and the 24h-UPRO, as well as between the renal parenchymal D, f values and the SCr. There was a significant positive correlation between all of the IVIM parameters and the eGFR. All of the IVIM parameters exhibited a significant negative correlation with the histopathological fibrosis score. CONCLUSION: IVIM-DWI shows great potential in the noninvasive assessment of renal fibrosis in CKD.

Longitudinal assessment of rabbit renal fibrosis induced by unilateral ureteral obstruction using two-dimensional susceptibility weighted imaging.

BACKGROUND: Previous studies indicated that two-dimensional-susceptibility weighted imaging (2D-SWI) could serve as a useful biomarker for differentiating the grade of liver fibrosis. PURPOSE: To evaluate the feasibility of 2D-SWI in the dynamic quantification of renal fibrosis in a rabbit model. STUDY TYPE: Longitudinal study. ANIMAL MODEL: Twenty-Four New Zealand White Rabbits including control group (n = 4); and renal fibrosis group (n = 20), by means of a unilateral ureteral obstruction (UUO) model. FIELD STRENGTH/SEQUENCE: The 3.0 T SWI using a 2D gradient-echo sequence. ASSESSMENT: The relative SWI signal ratio(r) of cortical and medulla (r = SIrenal /SImuscle ) was longitudinally assessed before ligation and on weeks 2, 4, 6, and 8 following ligation. Sirius Red staining was used to assess the degree of fibrosis in five high-power fields. STATISTICAL TESTS: The repeated measures of analysis of variance and linear regression analysis. RESULTS: Both the cortical and medullary r values were significantly higher in the UUO kidneys at week 2 compared with the kidneys before ligation. Over the course of UUO progression, significant changes occurred in the cortical and medullary r values in vivo and fibrosis scores in vitro (all P values < 0.05). The r values gradually decreased, while the fibrosis scores gradually increased over 8 weeks following ligation. The linear regression analysis showed a strong and significant correlation between cortical and medullary r values and the pathologic fibrosis scores (R2 = 0.91, 0.81, respectively). DATA CONCLUSION: The SWI sequence could provide a quantitative evaluation of renal fibrosis during UUO progression. LEVEL OF EVIDENCE: 2 Technical Efficacy: Stage 1 J. Magn. Reson. Imaging 2018;47:1572-1577.

Molecular validation of the precision-cut kidney slice (PCKS) model of renal fibrosis through assessment of TGF-ß1-induced Smad and p38/ERK signaling.

BACKGROUND: The precision-cut kidney slice (PCKS) model appears to be a useful ex vivo model of renal fibrosis. However, little in-depth molecular investigation on the PCKS model has been performed. Therefore, the aim of this study will be to investigate and validate the molecular validity of this model. METHODS: The PCKS model was constructed in male C57BL/6 mice. To induce renal fibrosis, PCKS were incubated in recombinant human TGF-ß1 for 48 h. Protein expression of phosphorylated Smad2 (p-Smad2, cytosolic and nuclear), Smad7, phosphorylated ERK1 (p-ERK1), phosphorylated ERK2 (p-ERK2), and phosphorylated p38 MAPK (p-p38 MAPK) was measured using Western blotting. To assess Smad2/3 heteromeric complex formation and phosphorylated Smad3 (p-Smad3) expression, immunoprecipitation was performed with an anti-Smad2 or an anti-Smad3 antibody, respectively, prior to Western blotting. RESULTS: p-Smad2 and p-Smad3 were significantly upregulated in the PCKS model relative to control (p<0.05). However, we found no significant difference in Smad7 expression between the PCKS model and control (p>0.05). The PCKS model demonstrated significantly greater Smad2/3 complex formation and nuclear translocation relative to control (p<0.05). The PCKS model showed significantly greater expression of p-ERK1, p-ERK2, and p-p38 MAPK relative to control (p<0.05). CONCLUSIONS: The PCKS model displays several well-established molecular markers of renal fibrosis. However, the PCKS model failed to display Smad7 downregulation and appears to display "over-activation" of p-Smad2 and p-Smad3 as well as "under-activation" of ERK1/2 and p38 MAPK signaling vis-à-vis the well-established in vivo unilateral ureteric obstruction model of renal fibrosis.