Comprehensive Proteomic Quantification of Bladder Stone Progression in a Cystinuric Mouse Model Using Data-Independent Acquisitions

Cystinuria is one of various disorders that cause biomineralization in the urinary system, including bladder stone formation in humans. It is most prevalent in children and adolescents and more aggressive in males. There is no cure, and only limited disease management techniques help to solubilize the stones. Recurrence, even after treatment, occurs frequently. Other than a buildup of cystine, little is known about factors involved in the formation, expansion, and recurrence of these stones. This study sought to define the growth of bladder stones, guided by micro-computed tomography imaging, and to profile dynamic stone proteome changes in a cystinuria mouse model. After bladder stones developed in vivo, they were harvested and separated into four developmental stages (sand, small, medium and large stone), based on their size. Data-dependent and data-independent acquisitions allowed deep profiling of stone proteomics. The proteomic signatures and pathways illustrated major changes as the stones grew. Stones initiate from a small nidus, grow outward, and show major enrichment in ribosomal proteins and factors related to coagulation and platelet degranulation, suggesting a major dysregulation in specific pathways that can be targeted for new therapeutic options.

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Lipoteichoic Acid Accelerates Bone Healing by Enhancing Osteoblast Differentiation and Inhibiting Osteoclast Activation in a Mouse Model of Femoral Defects

International Journal of Molecular Sciences ◽

10.3390/ijms21155550 ◽

2020 ◽

Vol 21 (15) ◽

pp. 5550

Author(s):

Chih-Chien Hu ◽

Chih-Hsiang Chang ◽

Yi-min Hsiao ◽

Yuhan Chang ◽

Ying-Yu Wu ◽

...

Keyword(s):

Mouse Model ◽

Bone Formation ◽

Bone Regeneration ◽

Bone Healing ◽

Lipoteichoic Acid ◽

Calcium Deposition ◽

Micro Computed Tomography ◽

Osteoclast Activation

Lipoteichoic acid (LTA) is a cell wall component of Gram-positive bacteria. Limited data suggest that LTA is beneficial for bone regeneration in vitro. Thus, we used a mouse model of femoral defects to explore the effects of LTA on bone healing in vivo. Micro-computed tomography analysis and double-fluorochrome labeling were utilized to examine whether LTA can accelerate dynamic bone formation in vivo. The effects of LTA on osteoblastogenesis and osteoclastogenesis were also studied in vitro. LTA treatment induced prompt bone bridge formation, rapid endochondral ossification, and accelerated healing of fractures in mice with femoral bone defects. In vitro, LTA directly enhanced indicators of osteogenic factor-induced MC3T3-E1 cell differentiation, including alkaline phosphatase activity, calcium deposition and osteopontin expression. LTA also inhibited osteoclast activation induced by receptor activator of nuclear factor-kappa B ligand. We identified six molecules that may be associated with LTA-accelerated bone healing: monocyte chemoattractant protein 1, chemokine (C-X-C motif) ligand 1, cystatin C, growth/differentiation factor 15, endostatin and neutrophil gelatinase-associated lipocalin. Finally, double-fluorochrome, dynamic-labeling data indicated that LTA significantly enhanced bone-formation rates in vivo. In conclusion, our findings suggest that LTA has promising bone-regeneration properties.

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SWI and phase imaging reveal intracranial calcifications in the P301L mouse model of human tauopathy

Magnetic Resonance Materials in Physics Biology and Medicine ◽

10.1007/s10334-020-00855-3 ◽

2020 ◽

Vol 33 (6) ◽

pp. 769-781 ◽

Cited By ~ 1

Author(s):

Ruiqing Ni ◽

Yvette Zarb ◽

Gisela A. Kuhn ◽

Ralph Müller ◽

Yankey Yundung ◽

...

Keyword(s):

Mouse Model ◽

Diamagnetic Susceptibility ◽

Ex Vivo ◽

Phase Imaging ◽

Phosphorylated Tau ◽

Gradient Echo ◽

Micro Computed Tomography ◽

Intracranial Calcifications ◽

Phase Images

Abstract Objective Brain calcifications are associated with several neurodegenerative diseases. Here, we describe the occurrence of intracranial calcifications as a new phenotype in transgenic P301L mice overexpressing four repeat tau, a model of human tauopathy. Materials and methods Thirty-six P301L mice (Thy1.2) and ten age-matched non-transgenic littermates of different ages were assessed. Gradient echo data were acquired in vivo and ex vivo at 7 T and 9.4 T for susceptibility-weighted imaging (SWI) and phase imaging. In addition, ex vivo micro-computed tomography (μCT) was performed. Histochemistry and immunohistochemistry were used to investigate the nature of the imaging lesions. Results SW images revealed regional hypointensities in the hippocampus, cortex, caudate nucleus, and thalamus of P301L mice, which in corresponding phase images indicated diamagnetic lesions. Concomitantly, µCT detected hyperdense lesions, though fewer lesions were observed compared to MRI. Diamagnetic susceptibility lesions in the hippocampus increased with age. The immunochemical staining of brain sections revealed osteocalcin-positive deposits. Furthermore, intra-neuronal and vessel-associated osteocalcin-containing nodules co-localized with phosphorylated-tau (AT8 and AT100) in the hippocampus, while vascular osteocalcin-containing nodules were detected in the thalamus in the absence of phosphorylated-tau deposition. Discussion SWI and phase imaging sensitively detected intracranial calcifications in the P301L mouse model of human tauopathy.

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In-vivo lung fibrosis staging in a bleomycin-mouse model: a new micro-CT guided densitometric approach

Scientific Reports ◽

10.1038/s41598-020-71293-3 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Laura Mecozzi ◽

Martina Mambrini ◽

Francesca Ruscitti ◽

Erica Ferrini ◽

Roberta Ciccimarra ◽

...

Keyword(s):

Pulmonary Fibrosis ◽

Mouse Model ◽

Lung Fibrosis ◽

Hounsfield Unit ◽

Lung Parenchyma ◽

Micro Ct ◽

Micro Computed Tomography ◽

Ct Guided ◽

Fibrosis Staging

Abstract Although increasing used in the preclinical testing of new anti-fibrotic drugs, a thorough validation of micro-computed tomography (CT) in pulmonary fibrosis models has not been performed. Moreover, no attempts have been made so far to define density thresholds to discriminate between aeration levels in lung parenchyma. In the present study, a histogram-based analysis was performed in a mouse model of bleomycin (BLM)-induced pulmonary fibrosis by micro-CT, evaluating longitudinal density changes from 7 to 21 days after BLM challenge, a period representing the progression of fibrosis. Two discriminative densitometric indices (i.e. 40th and 70th percentiles) were extracted from Hounsfield Unit density distributions and selected for lung fibrosis staging. The strong correlation with histological findings (rSpearman = 0.76, p < 0.01) confirmed that variations in 70th percentile could reflect a pathological lung condition and estimate the effect of antifibrotic treatments. This index was therefore used to define lung aeration levels in mice distinguishing in hyper-inflated, normo-, hypo- and non-aerated pulmonary compartments. A retrospective analysis performed on a large cohort of mice confirmed the correlation between the proposed preclinical density thresholds and the histological outcomes (rSpearman = 0.6, p < 0.01), strengthening their suitability for tracking disease progression and evaluating antifibrotic drug candidates.

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Tau deposition is associated with imaging patterns of tissue calcification in the P301L mouse model of human tauopathy

10.1101/851915 ◽

2019 ◽

Cited By ~ 1

Author(s):

Ruiqing Ni ◽

Yvette Zarb ◽

Gisela A. Kuhn ◽

Ralph Müller ◽

Yankey Yundung ◽

...

Keyword(s):

Mouse Model ◽

Ex Vivo ◽

Intracranial Calcification ◽

Phosphorylated Tau ◽

Micro Computed Tomography ◽

Brain Calcification ◽

Phase Images ◽

Magnetic Resonance Imaging Mri ◽

Immunohistochemical Evidence

AbstractBrain calcification is associated with several neurodegenerative proteinopathies. Here, we report a new phenotype of intracranial calcification in transgenic P301L mice overexpressing 4 repeat tau. P301L mice (Thy1.2) of 3, 5, 9 and 18-25 months-of-age and age-matched non-transgenic littermates were assessed using in vivo/ex vivo magnetic resonance imaging (MRI) with a gradient recalled echo sequence and micro computed tomography (μCT). Susceptibility weighted images computed from the gradient recalled echo data revealed regional hypointensities in the hippocampus, cortex, caudate nucleus and thalamus of P301L mice, which in corresponding phase images indicated diamagnetic lesions. Concomitantly, µCT detected hyperdense lesions. Occurrence of diamagnetic susceptibility lesions in the hippocampus, increased with age. Immunochemical staining of brain sections revealed bone protein-positive deposits. Furthermore, intra-neuronal and vessel-associated protein-containing nodules co-localized with phosphorylated-tau (AT8 and AT100) in the hippocampus. Protein-containing nodules were detected also in the thalamus in the absence of phosphorylated-tau deposition. In contrast, osteocalcin-containing nodules were vessel-associated, indicating ossified vessels, in the thalamus in absence of phosphorylated-tau. In summary, MRI and µCT demonstrated imaging pattern of intracranial calcification, concomitant with immunohistochemical evidence of formation of protein deposits containing bone proteins along with phosphorylated-tau in the P301L mouse model of human tauopathy. The P301L mouse model may thus serve as a future model to study the pathogenesis of brain calcifications in tauopathies.

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