Pten regulates collagen fibrillogenesis by fibroblasts through SPARC

Jones, Caitlin E.; Sharick, Joe T.; Colbert, Sheila E.; Shukla, Vasudha; Zent, Joshua; Ostrowski, Michael C.; Ghadiali, Samir N.; Sizemore, Steven T.; Leight, Jennifer L.

doi:10.1371/journal.pone.0245653

Cited by 13 publications

(13 citation statements)

References 64 publications

(95 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Thus, the distinction between secretion of soluble collagen and fibril assembly is important in studies of disease, in which simple measurement of collagen expression or secretion does not equate to fibrogenesis. This distinction between collagen secretion and assembly was also suggested in a recent study on Pten- knockout mammary fibroblasts, where SPARC affects collagen fibrillogenesis and not secretion, acting through fibronectin 49 . Of note, whilst SPARC and fibronectin were detected in our previously-reported time-series mass spectrometry analyses (in the SLDOC soluble fraction and high-salt insoluble fractions respectively), they were not circadian clock rhythmic 13 .…”

Section: Discussionsupporting

confidence: 57%

Endocytic recycling is central to circadian collagen fibrillogenesis and disrupted in fibrosis

Chang

Pickard

Garva

et al. 2021

Preprint

View full text Add to dashboard Cite

Collagen fibrils are the principal supporting elements in vertebrate tissues. They account for 25% of total protein mass, exhibit a broad range of size and organisation depending on tissue and stage of development, and can be under circadian clock control. Here we show that the remarkable dynamic pleomorphism of collagen fibrils is underpinned by a mechanism that distinguishes between collagen secretion and initiation of fibril assembly, at the plasma membrane. Collagen fibrillogenesis occurring at the plasma membrane requires vacuolar protein sorting (VPS) 33b (which is under circadian clock control), collagen-binding integrin-α11 subunit, and is reduced when endocytosis is inhibited. Fibroblasts lacking VPS33b secrete soluble collagen without assembling fibrils, whereas constitutive over-expression of VPS33b increases fibril number with loss of fibril rhythmicity. In conclusion, our study has identified the mechanism that switches secretion of collagen (without forming new fibrils) to new collagen fibril assembly, at the plasma membrane.

show abstract

Section: Discussionsupporting

confidence: 57%

Endocytic recycling is central to circadian collagen fibrillogenesis and disrupted in fibrosis

Chang

Pickard

Garva

et al. 2021

Preprint

View full text Add to dashboard Cite

show abstract

“…MiR-29c-5p might also target PTEN. Low stromal PTEN expression is associated with multiple cancers, but also with high mammographic density through collagen deposition [ 44 ]. Murine PTEN knockout models demonstrate more collagen secretion, fiber formation and increased expression of the ECM regulator SPARC [ 44 ].…”

Section: Discussionmentioning

confidence: 99%

“…Low stromal PTEN expression is associated with multiple cancers, but also with high mammographic density through collagen deposition [ 44 ]. Murine PTEN knockout models demonstrate more collagen secretion, fiber formation and increased expression of the ECM regulator SPARC [ 44 ]. Furthermore, a mouse model with fibroblast-specific PTEN deletion resulted in an alteration of collagen organization, a promotion of collagen orientation in surrounding tumors, and an enhancement of highly aligned matrices when comparing with a wildtype model [ 45 ].…”

Section: Discussionmentioning

confidence: 99%

Elevated miR-29c-5p Expression in Nipple Aspirate Fluid Is Associated with Extremely High Mammographic Breast Density

Vissers

Piek

Patuleia

et al. 2022

Cancers

View full text Add to dashboard Cite

High mammographic density (MD) is associated with an increased risk of breast cancer, however the underlying mechanisms are largely unknown. This research aimed to identify microRNAs (miRNAs) that play a role in the development of extremely dense breast tissue. In the discovery phase, 754 human mature miRNAs were profiled in 21 extremely high MD- and 20 very low MD-derived nipple aspirate fluid (NAF) samples from healthy women. In the validation phase, candidate miRNAs were assessed in a cohort of 89 extremely high MD and 81 very low MD NAF samples from healthy women. Independent predictors of either extremely high MD or miRNA expression were identified by logistic regression and linear regression analysis, respectively. mRNA targets and pathways were identified through miRTarBase, TargetScan, and PANTHER pathway analysis. Statistical analysis identified four differentially expressed miRNAs during the discovery phase. During the validation, linear regression (p = 0.029; fold change = 2.10) and logistic regression (p = 0.048; odds ratio = 1.38) showed that hsa-miR-29c-5p was upregulated in extremely high MD-derived NAF. Identified candidate mRNA targets of hsa-miR-29c-5p are CFLAR, DNMT3A, and PTEN. Further validation and exploration of targets and downstream pathways of has-miR-29c-5p will provide better insight into the processes involved in the development of high MD and in the associated increased risk of breast cancer.

show abstract

“…To eliminate these costly and time-consuming immunostaining steps and to enable visualization of the matrix for the screening platform, cells were cultured with exogenous fluorescein-labeled fibronectin. Previous studies have demonstrated the incorporation of fluorescent fibronectin into cell-derived matrices (74,75), and the addition of exogenous fluorescent fibronectin has previously been used to study fibronectin fibrillogenesis (62,76). Furthermore, fibronectin is oriented in an equivalent manner as collagen and is the most prevalent protein expressed in CDM (1).…”

Section: Miniaturization and Streamlining Of The Matrix Organization ...mentioning

confidence: 99%

“…In vivo ablation of Pten (Fsp-Cre;Pten loxP/loxP ) in fibroblasts of mouse mammary tissue led to increased collagen deposition, collagen alignment, and tumorigenesis in the presence of the Neu oncogene (58,63). In vitro, CDM produced by Pten null (Pten -/-) murine mammary fibroblasts results in a highly organized, aligned matrix compared to the largely disorganized matrix produced by Pten intact (wild type) fibroblasts (63,75). As such, these cells are well suited for screening compounds that can 'normalize' a highly aligned matrix.…”

Section: Kinase Inhibitor Screen Identifies Novel Regulators Of Matri...mentioning

confidence: 99%

A Miniaturized Screening Platform to Identify Novel Regulators of Extracellular Matrix Alignment

Jones

Sharick

Sizemore

et al. 2022

Cancer Research Communications

Self Cite

View full text Add to dashboard Cite

Extracellular matrix alignment contributes to metastasis in a number of cancers and is a known prognostic stromal factor; however, the mechanisms controlling matrix organization remain unclear. Cancer-associated fibroblasts (CAF) play a critical role in this process, particularly via matrix production and modulation of key signaling pathways controlling cell adhesion and contractility. Stroma normalization, as opposed to elimination, is a highly sought strategy, and screening for drugs that effectively alter extracellular matrix (ECM) alignment is a practical way to identify novel CAF-normalizing targets that modulate ECM organization. To meet this need, we developed a novel high-throughput screening platform in which fibroblast-derived matrices were produced in 384-well plates, imaged with automated confocal microscopy, and analyzed using a customized MATLAB script. This platform is a technical advance because it miniaturizes the assay, eliminates costly and time-consuming experimental steps, and streamlines data acquisition and analysis to enable high-throughput screening applications. As a proof of concept, this platform was used to screen a kinase inhibitor library to identify modulators of matrix alignment. A number of novel potential regulators were identified, including several receptor tyrosine kinases (c-MET, tropomyosin receptor kinase 1 (NTRK1), HER2/ERBB2) and the serine/threonine kinases protein kinase A, C, and G (PKA, PKC, and PKG) . The expression of these regulators was analyzed in publicly available patient datasets to examine the association between stromal gene expression and patient outcomes.

show abstract

Pten regulates collagen fibrillogenesis by fibroblasts through SPARC

Cited by 13 publications

References 64 publications

Endocytic recycling is central to circadian collagen fibrillogenesis and disrupted in fibrosis

Endocytic recycling is central to circadian collagen fibrillogenesis and disrupted in fibrosis

Elevated miR-29c-5p Expression in Nipple Aspirate Fluid Is Associated with Extremely High Mammographic Breast Density

A Miniaturized Screening Platform to Identify Novel Regulators of Extracellular Matrix Alignment

Contact Info

Product

Resources

About