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Bone morphogenetic protein 4 (BMP-4) is a member of the highly conserved transforming growth factor 尾 (TGF-尾) superfamily. Mature BMP-4 is a disulfide-linked homodimeric protein consisting of two 116-amino-acid residue subunits, and is generated by the proteolytic removal of the signal peptide and propeptides (Xiao et al.). BMP-4 binds to type I and type II receptors on cells. This binding results in the phosphorylation of receptor 1, which in turn results in the phosphorylation of Smad proteins, which then go on to act as transcription factors (Zhang et al.). BMPs have been shown to be key regulators of embryogenesis and are known to play a role in the growth and differentiation of various cell types, including embryonic stem (ES) cells, induced pluripotent stem (iPS) cells, mesenchymal cells, epithelial cells, hematopoietic cells, and neuronal cells (Chadwick et al.; Graham et al.; Jones et al.; Lengerke et al.; Zhang et al.).
(A) The biological activity of Human Recombinant BMP-4 was tested by its ability to induce alkaline phosphatase production in ATDC-5 cells. Alkaline phosphatase production was measured using a fluorometric assay method. The EC50 is defined as the effective concentration of the growth factor at which alkaline phosphatase activity is at 50% of maximum. The EC50 in the above example is 3.99 ng/mL.
(B) Human Recombinant BMP-4 was resolved with SDS-PAGE under reducing (+) conditions and non-reducing (-) conditions and visualized by Coomassie Blue staining. The predicted molecular mass is 13.1 kDa (monomer) or 26.2 kDa (dimer), but these migrate to an apparent molecular mass of 20 kDa or 34 kDa, respectively, due to variable glycosylation.
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Rab7 inhibitor enhances stem cell differentiation into keratinocyte-like cells with anti-inflammatory properties
R. Alghazali et al.
Frontiers in Immunology 2025 May
Abstract
Management of difficult-to-heal skin wounds presents a significant clinical challenge, particularly when associated with inflammatory skin disorders. The differentiation of stem cells into keratinocyte-like cells has been explored as a potential regenerative therapy. Ras-related protein (Rab) 7, a key regulator of membrane trafficking, influences the activity of several growth factors. In this study, the competitive Rab7 inhibitor, CID-1067700, was investigated for the differentiation of adipose-derived stem cell into keratinocyte-like cells. This treatment upregulated several epidermal markers, including P63, cytokeratin 5 and 14 and filaggrin, while downregulated the stem cell marker, vimentin. Microarray data showed upregulation of the anti-inflammatory gene HMOX-1, coupled with the downregulation of various inflammation-related pathways, such as TNF, chemokine, AGE-RAGE and IL-17 signalling cascades, as well as cytokine-cytokine receptor interaction pathways. Gene set enrichment analysis predicted Ehmt2, an epigenetic regulator, to be the top activated upstream regulator, enhancing the transcriptional activity. Protein array analysis showed reduced secretion of several inflammatory cytokines, including IL-1伪, IL-8, IL-17A, and IL-32, while enhancing the secretion of the epidermal growth factor. Our findings provide preliminary evidence that CID-1067700, as an additive to the differentiation media, not only enhances stem cell differentiation into keratinocyte-like cells, but also improves their anti-inflammatory properties. These combined regenerative and anti-inflammatory properties may offer significant therapeutic potential for patients with chronic skin wounds, particularly those with underlying inflammatory conditions.
