Interfering with stem cell-specific gatekeeper functions controls tumour initiation and malignant progression of skin tumours.

Epithelial cancer constitutes a major clinical challenge and molecular mechanisms underlying the process of tumour initiation are not well understood. Here we demonstrate that hair follicle bulge stem cells (SCs) give rise to well-differentiated sebaceous tumours and show that SCs are not only crucial in tumour initiation, but are also involved in tumour plasticity and heterogeneity. Our findings reveal that SC-specific expression of mutant Lef1, which mimics mutations found in human sebaceous tumours, drives sebaceous tumour formation. Mechanistically, we demonstrate that mutant Lef1 abolishes p53 activity in SCs. Intriguingly, mutant Lef1 induces DNA damage and interferes with SC-specific gatekeeper functions normally protecting against accumulations of DNA lesions and cell loss. Thus, normal control of SC proliferation is disrupted by mutant Lef1, thereby allowing uncontrolled propagation of tumour-initiating SCs. Collectively, these findings identify underlying molecular and cellular mechanisms of tumour-initiating events in tissue SCs providing a potential target for future therapeutic strategies.

Mitochondrial function in murine skin epithelium is crucial for hair follicle morphogenesis and epithelial-mesenchymal interactions.

Here, we studied how epithelial energy metabolism impacts overall skin development by selectively deleting intraepithelial mtDNA in mice by ablating a key maintenance factor (Tfam(EKO)), which induces loss of function of the electron transport chain (ETC). Quantitative (immuno)histomorphometry demonstrated that Tfam(EKO) mice showed significantly reduced hair follicle (HF) density and morphogenesis, fewer intrafollicular keratin15+ epithelial progenitor cells, increased apoptosis, and reduced proliferation. Tfam(EKO) mice also displayed premature entry into (aborted) HF cycling by apoptosis-driven HF regression (catagen). Ultrastructurally, Tfam(EKO) mice exhibited severe HF dystrophy, pigmentary abnormalities, and telogen-like condensed dermal papillae. Epithelial HF progenitor cell differentiation (Plet1, Lrig1 Lef1, and ß-catenin), sebaceous gland development (adipophilin, Scd1, and oil red), and key mediators/markers of epithelial-mesenchymal interactions during skin morphogenesis (NCAM, versican, and alkaline phosphatase) were all severely altered in Tfam(EKO) mice. Moreover, the number of mast cells, major histocompatibility complex class II+, or CD11b+ immunocytes in the skin mesenchyme was increased, and essentially no subcutis developed. Therefore, in contrast to their epidermal counterparts, pilosebaceous unit stem cells depend on a functional ETC. Most importantly, our findings point toward a frontier in skin biology: the coupling of HF keratinocyte mitochondrial function with the epithelial-mesenchymal interactions that drive overall development of the skin and its appendages.

Indian hedgehog controls proliferation and differentiation in skin tumorigenesis and protects against malignant progression.

Mutations in the hedgehog pathway drive the formation of tumors in many different organs, including the development of basal cell carcinoma in the skin. However, little is known about the role of epidermal Indian hedgehog (Ihh) in skin physiology. Using mouse genetics, we identified overlapping and distinct functions of Ihh in different models of epidermal tumorigenesis. Epidermal deletion of Ihh resulted in increased formation of benign squamous papilloma. Strikingly, Ihh-deficient mice showed an increase in malignant squamous cell carcinoma and developed lung and lymph node metastases. In a sebaceous gland tumor model, Ihh deficiency inhibited tumor cell differentiation. More mechanistically, IHH stimulated cell proliferation by activating the transcription factor GLI2 in human keratinocytes and human tumors. Thus, our results uncover important functions for Ihh signaling in controlling proliferation, differentiation, malignant progression, and metastasis of epithelial cancer, establishing Ihh as a gatekeeper for controlling the grade of tumor malignancy.

Is it necessary to penalize impulsive noise +5 dB due to higher risk of hearing damage?

It is studied whether the +5 dB penalty for impulsiveness established by ISO 1999:1990 accounts for a higher risk of noise-induced hearing loss. A total of 16 normal-hearing human subjects were exposed for 10 min to two types of binaural industrial-recordings: (1) a continuous broad-band noise normalized to L(EX,8 h)=80 dBA and (2) the combination of the previous stimulus with an impulsive noise normalized to L(EX,8 h)=75+5(db penalty)=80 dBA (peak level 117 dBC and repetition rate of 0.5 impacts per second). Distortion product otoacoustic emissions (DPOAEs) were measured in a broad frequency range before and in the following 90 min after the exposure. The group results show that the continuous exposure had a bigger impact on DPOAE levels, with a maximum DPOAE shift of approximately 5 dB in the frequency range of 2-3.15 kHz during the first 10 min of the recovery. No evident DPOAE shift is seen for the impulsive + continuous stimulus. The results indicate that the penalty overestimated the effects on DPOAE levels and support the concept that the risk of hearing loss from low-level impulses may be predicted on an equal-energy basis.

Recovery of distortion-product otoacoustic emissions after a 2-kHz monaural sound-exposure in humans: effects on fine structures.

A better understanding of the vulnerability of the fine structures of distortion-product otoacoustic emissions (DPOAEs) after acoustic overexposure may improve the knowledge about DPOAE generation, cochlear damage, and lead to more efficient diagnostic tools. It is studied whether the DPOAE fine structures of 16 normal-hearing human subjects are systematically affected after a moderate monaural sound-exposure of 10 min to a 2-kHz tone normalized to an exposure level L(EX,8h) of 80 dBA. DPOAEs were measured before and in the following 70 min after the exposure. The experimental protocol allowed measurements with high time and frequency resolution in a 1/3-octave band centered at 3 kHz. On average, DPOAE levels were reduced approximately 5 dB in the entire measured frequency-range. Statistically significant differences in pre- and post-exposure DPOAE levels were observed up to 70 min after the end of the sound exposure. The results show that the effects on fine structures are highly individual and no systematic change was observed.

Overexposure effects of a 1-kHz tone on the distortion product otoacoustic emission in humans.

The effects of overexposure on the properties of distortion product otoacoustic emissions (DPOAEs) are investigated. In total, 39 normal-hearing humans were monaurally exposed to a 1-kHz tone lasting for 3 min at an equivalent threshold sound-pressure level of 105.5 dB. The effects of overexposure were studied in two experiments (1) on the broadband DPOAE and (2) on the DPOAE fine structure, measured using a higher frequency resolution in a narrower frequency range. The obtained DPOAE shifts were compared to temporary threshold shift (TTS) obtained after a similar exposure. Similarities between DPOAE shifts and TTS were found in the affected frequency range and the time course of recovery. The amount of TTS was higher in the early recovery time (1-4-min postexposure), but similar to the DPOAE shift (even in absolute terms) at later recovery times (5-20-min postexposure). The DPOAE fine structure was not systematically changed after the exposure.

Distortion product otoacoustic emission of symphony orchestra musicians before and after rehearsal.

The 2f1-f2 distortion product otoacoustic emission (DPOAE) and hearing levels are obtained for 12 normal-hearing symphony orchestra musicians both before and after their rehearsal. The DPOAE fine structures are determined and analyzed according to the character and prevalence of ripples. Hearing levels, DPOAE levels, and DPOAE fine structures before and after rehearsal are similar, indicating that no or marginal temporary change of the state of hearing were caused by the exposure. The data were further compared to similar data for occupationally nonexposed subjects, one group which was age and gender matched, and other two groups of younger individuals (one group with better hearing levels than the other). The data for the age and gender matched group compared well with the musicians data (and the data for the group of better-hearing younger individuals). In general, the analyses of hearing thresholds and DPOAE data thus lead to the same conclusions concerning the state of hearing.

Distortion product otoacoustic emission fine structure analysis of 50 normal-hearing humans.

When distortion product otoacoustic emissions (DPOAEs) are measured with a high-frequency resolution, the DPOAE shows quasi-periodic variations across frequency, called DPOAE fine structure. In this study the DPOAE fine structure is determined for 50 normal-hearing humans using fixed primary levels of L1/L2 = 65/45 dB. An algorithm is developed, which characterizes the fine structure ripples in terms of three parameters: ripple spacing, ripple height, and ripple prevalence. The characteristic patterns of fine structure can be found in the DPOAE of all subjects, though the DPOAE fine structure characteristics are individual and vary from subject to subject. On average the ripple spacing decreases with increasing frequency from 1/8 oct at 1 kHz to 3/32 oct at 5 kHz. The ripple prevalence is two to three ripples per 1/3 oct, and ripple heights of up to 32 dB could be detected. The 50 normal-hearing subjects were divided into two groups, the subjects of group A having slightly better hearing levels than subjects of group B. The subjects of group A have significantly higher DPOAE levels. The overall prevalence of fine structure ripples do not differ between the two groups, but are higher and narrower for subjects of group B than for group A.