Stem cell markers (cytokeratin 15, CD34 and nestin) in primary scarring and nonscarring alopecia.

BACKGROUND: Although the pathogenesis of most primary scarring alopecias is poorly understood, recent studies implicate the bulge region as a possible target. OBJECTIVES: To corroborate these results, we ascertained involvement of follicular bulge stem cells using a panel of antibodies that putatively targeted the same. METHODS: Antibodies used included anticytokeratin (CK) 15, CD34 and nestin on vertical and horizontal tissue sections of 50 cases of scarring and 34 cases of nonscarring alopecia. RESULTS: Comparing expression of these markers in scarring vs. nonscarring alopecia, CK15 was noted in the follicular bulge region in 23 of 43 (53%) vs. 27 of 27 (100%) cases and in the peripheral layer of the outer root sheath (ORS) (upper two-thirds of the follicle) in 50 of 50 (100%) vs. 34 of 34 (100%) cases; CD34 was noted in the peripheral layer of the ORS (below pilar muscle attachment) in 24 of 35 (69%) vs. 18 of 18 (100%) cases; and nestin was noted in the infundibular region in 18 of 46 (39%) vs. seven of 32 (22%) cases and in the inner aspect of the ORS (below pilar muscle attachment) in eight of 31 (26%) vs. 23 of 23 (100%) cases. CONCLUSIONS: Our findings of differential follicular localization of stem cells underscore follicular progenitor cell heterogeneity and suggest the target in scarring alopecia is not merely follicular bulge stem cells but involves stem cells in the inner and outer aspect of the ORS. Enhanced expression of nestin in the infundibular region in scarring alopecia indicates availability of an accessible, in vivo niche of potential utility as an autologous source of stem cells for therapeutic application.

RAS and RAF mutations in banal melanocytic aggregates contiguous with primary cutaneous melanoma: clues to melanomagenesis.

BACKGROUND: Distinguishing banal melanocytic aggregates contiguous with malignant melanoma can be a histological challenge but is essential because of the potential for a spurious Breslow measurement. OBJECTIVES: Our aim was to ascertain whether the histological distinction between the two relates to differences in the prevalence of mutations in genes significant in melanomagenesis. METHODS: Mutations in BRAF codon 600, NRAS1 codons 12/13, NRAS2 codons 60/61 and KRAS codons 12/13 were ascertained in 18 cases of primary cutaneous malignant melanoma contiguous with banal melanocytic aggregates using laser capture microdissection. RESULTS: Overall, 12 of 18 cases (67%) exhibited a mutation in at least one gene. BRAF V600E appeared to be the most commonly mutated gene in both the melanocytic aggregate (seven of 18, 39%) and the melanoma (four of 18, 22%). Both populations demonstrated a similar BRAF genomic profile in 11 of 18 cases (61%) (two BRAF V600E, nine BRAF-WT), a similar KRAS genomic profile in 14 of 18 cases (78%) (one KRAS G12V, 13 KRAS-WT) and a similar NRAS2 genomic profile in 14 of 18 cases (all WT). Of interest, we noted a relatively high prevalence of KRAS mutations (five of 18, 28%). The frequency of KRAS mutations in the melanocytic aggregate (five of 18, 28%) was second to BRAF V600E, while in melanoma, the frequency was also second to BRAF V600E but equalled that of NRAS2 (1 of 18, 6%). No NRAS1 mutations were observed. BRAF and RAS mutations appeared to be mutually exclusive with only three of 18 cases (17%) demonstrating a mutation in both genes (melanocytic aggregate only). CONCLUSIONS: Our findings hint towards the interpretation of banal melanocytic aggregates serving as precursor lesions.