Proteome array identification of bioactive soluble proteins/peptides in Matrigel: relevance to stem cell responses

Talbot, Neil C.; Caperna, Thomas J.

doi:10.1007/s10616-014-9727-y

Cited by 61 publications

(57 citation statements)

References 43 publications

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“…Certain laminin isoforms, fibronectin and collagen type-IV, are critical for the development of the neural crest [45]. NB cells migrate extensively in Matrigel, which contains these ECM molecules [27], which is in line with our findings.…”

Section: Matrigel Is the Preferred Substratum For Nb Invasionsupporting

confidence: 91%

Neuroblastoma Invasion Strategies Are Regulated by the Extracellular Matrix

Gavin¹,

Geerts²,

Cavanagh

et al. 2021

Cancers

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Neuroblastoma is a paediatric malignancy of the developing sympathetic nervous system. About half of the patients have metastatic disease at the time of diagnosis and a survival rate of less than 50%. Our understanding of the cellular processes promoting neuroblastoma metastases will be facilitated by the development of appropriate experimental models. In this study, we aimed to explore the invasion of neuroblastoma cells and organoids from patient-derived xenografts (PDXs) grown embedded in 3D extracellular matrix (ECM) hydrogels by time-lapse microscopy and quantitative image analysis. We found that the ECM composition influenced the growth, viability and local invasion of organoids. The ECM compositions induced distinct cell behaviours, with Matrigel being the preferred substratum for local organoid invasion. Organoid invasion was cell line- and PDX-dependent. We identified six distinct phenotypes in PDX-derived organoids. In contrast, NB cell lines were more phenotypically restricted in their invasion strategies, as organoids isolated from cell line-derived xenografts displayed a broader range of phenotypes compared to clonal cell line clusters. The addition of FBS and bFGF induced more aggressive cell behaviour and a broader range of phenotypes. In contrast, the repression of the prognostic neuroblastoma marker, MYCN, resulted in less aggressive cell behaviour. The combination of PDX organoids, real-time imaging and the novel 3D culture assays developed herein will enable rapid progress in elucidating the molecular mechanisms that control neuroblastoma invasion.

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Section: Matrigel Is the Preferred Substratum For Nb Invasionsupporting

confidence: 91%

Neuroblastoma Invasion Strategies Are Regulated by the Extracellular Matrix

Gavin¹,

Geerts²,

Cavanagh

et al. 2021

Cancers

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“…It has been shown that Matrigel contained various biologically active proteins, such as IGF1, TGFβ, EGF, bFGF, and PDGF, that could increase the frequency of tumor xenograft (20). Hence, to identify the stimuli, we selected 11 cytokines that were enriched in Matrigel (SI Appendix, Table S2) (20,21). These factors were divided into three groups according to their different bioactivities, such as promoting cell proliferation, stimulating angiogenesis, and regulating cell stemness, and administrated through tail i.v.…”

Section: Resultsmentioning

confidence: 99%

Expansion of cancer stem cell pool initiates lung cancer recurrence before angiogenesis

Yang

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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Angiogenesis is essential in the early stage of solid tumor recurrence, but how a suspensive tumor is reactivated before angiogenesis is mostly unknown. Herein, we stumble across an interesting phenomenon that s.c. xenografting human lung cancer tissues can awaken the s.c. suspensive tumor in nude mice. We further found that a high level of insulin-like growth factor 1 (IGF1) was mainly responsible for triggering the transition from suspensive tumor to progressive tumor in this model. The s.c. suspensive tumor is characterized with growth arrest, avascularity, and a steady-state level of proliferating and apoptotic cells. Intriguingly, CD133 lung cancer stem cells (LCSCs) are highly enriched in suspensive tumor compared with progressive tumor. Mechanistically, high IGF1 initiates LCSCs self-renewal from asymmetry to symmetry via the activation of a PI3K/Akt/β-catenin axis. Next, the expansion of LCSC pool promotes angiogenesis by increasing the production of CXCL1 and PlGF in CD133 LCSCs, which results in lung cancer recurrence. Clinically, a high level of serum IGF1 in lung cancer patients after orthotopic lung cancer resection as an unfavorable factor is strongly correlated with the high rate of recurrence and indicates an adverse progression-free survival. Vice versa, blocking IGF1 or CXCL1/PlGF with neutralizing antibodies can prevent the reactivation of a suspensive tumor induced by IGF1 stimulation in the mouse model. Collectively, the expansion of LCSC pool before angiogenesis induced by IGF1 is a key checkpoint during the initiation of cancer relapse, and targeting serum IGF1 may be a promising treatment for preventing recurrence in lung cancer patients.

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“…Commercial Matrigel is widely used in human stem cell cultures by mimicking the extracellular matrix (ECM) [55], and it is known to contain components other than structural ECM proteins, such as growth factors and MMPs 2 and 9 [56], or even intracellular or membrane proteins [55]. Talbot et al [57] demonstrated by semi-quantitative analyses, with capture antibody arrays, the presence of various secreted or soluble proteins and peptides classified as growth factors, chemokines and other proteins with biological activity in four different lots from commercial Matrigel, including the Vascular Endothelial Growth Factor (VEGF), chemokine MCP-1 and peptide C5a. However, these results did not answer some questions about the biological activity of proteins or peptides present in Matrigel, which could impact experiments in vitro involving cell culture.…”

Section: Discussionmentioning

confidence: 99%

Inflammatory Reaction Induced by Two Metalloproteinases Isolated from Bothrops atrox Venom and by Fragments Generated from the Hydrolysis of Basement Membrane Components

Almeida

Freitas-de-Sousa

Colombini

et al. 2020

Toxins

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Snake venom metalloproteinases (SVMPs) play an important role in local tissue damage of snakebite patients, mostly by hydrolysis of basement membrane (BM) components. We evaluated the proinflammatory activity of SVMPs Atroxlysin-Ia (ATXL) and Batroxrhagin (BATXH) from Bothrops atrox venom and their hydrolysis products of Matrigel. BALB/c mice were injected with SVMPs (2 μg), for assessment of paw edema and peritoneal leukocyte accumulation. Both SVMPs induced edema, representing an increase of ~70% of the paw size. Leukocyte infiltrates reached levels of 6 × 106 with ATXL and 5 × 106 with BATXH. TNF-α was identified in the supernatant of BATXH—or venom-stimulated MPAC cells. Incubation of Matrigel with the SVMPs generated fragments, including peptides from Laminin, identified by LC–MS/MS. The Matrigel hydrolysis peptides caused edema that increased 30% the paw size and promoted leukocyte accumulation (4–5 × 106) to the peritoneal cavity, significantly higher than Matrigel control peptides 1 and 4 h after injection. Our findings suggest that ATXL and BATXH are involved in the inflammatory reaction observed in B. atrox envenomings by direct action on inflammatory cells or by releasing proinflammatory peptides from BM proteins that may amplify the direct action of SVMPs through activation of endogenous signaling pathways.

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Proteome array identification of bioactive soluble proteins/peptides in Matrigel: relevance to stem cell responses

Cited by 61 publications

References 43 publications

Neuroblastoma Invasion Strategies Are Regulated by the Extracellular Matrix

Neuroblastoma Invasion Strategies Are Regulated by the Extracellular Matrix

Expansion of cancer stem cell pool initiates lung cancer recurrence before angiogenesis

Inflammatory Reaction Induced by Two Metalloproteinases Isolated from Bothrops atrox Venom and by Fragments Generated from the Hydrolysis of Basement Membrane Components

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