Adele Haimovic

microRNA-214 contributes to melanoma tumour progression through suppression of TFAP2C

Malignant melanoma is fatal in its metastatic stage. It is therefore essential to unravel the molecular mechanisms that govern disease progression to metastasis. MicroRNAs (miRs) are endogenous non‐coding RNAs involved in tumourigenesis. Using a melanoma progression model, we identified a novel pathway controlled by miR‐214 that coordinates metastatic capability. Pathway components include TFAP2C, homologue of a well‐established melanoma tumour suppressor, the adhesion receptor ITGA3 and multiple surface molecules. Modulation of miR‐214 influences in vitro tumour cell movement and survival to anoikis as well as extravasation from blood vessels and lung metastasis formation in vivo. Considering that miR‐214 is known to be highly expressed in human melanomas, our data suggest a critical role for this miRNA in disease progression and the establishment of distant metastases.

Deletion of PTENP1 Pseudogene in Human Melanoma

Dear Editor, The aberrant activation of the PI3K/AKT pathway in melanoma is known to be caused by genomic deletion, promoter methylation or loss-of-function mutations of phosphatase and tensin homolog on chromosome 10 (PTEN) (see ref. (Madhunapantula and Robertson, 2009) for a review). More recently, PTEN protein abundance was shown to be decreased at the post-transcriptional level by a complex microRNA network (He, 2010). The processed pseudogene of PTEN, PTENP1, is a modulator of the interaction between PTEN mRNA and PTEN-targeting microRNAs (Poliseno et al., 2010). PTENP1 shows extensive sequence similarity with PTEN. The high level of conservation in the most upstream region of the 3’UTR allows PTENP1 mRNA to be bound by many of the microRNA families that bind PTEN mRNA (Figure 1 and Supplemental Text S1). Therefore, PTENP1 can protect PTEN from microRNA-mediated downregulation. Consistent with its activity as a decoy for PTEN-targeting microRNAs, PTENP1 is a bona fide tumor suppressor gene that causes growth inhibition in prostate cancer cells and undergoes deletion in various human malignancies (Poliseno et al, 2010). Figure 1 PTEN- and PTENP1-targeting microRNAs As recently pointed out (Chen, 2010), the post-transcriptional regulation of PTEN levels in melanoma has not been studied yet and the status of PTENP1 on 9p13 is unknown. Here, we report the analysis of PTENP1 locus in human melanoma cell lines and tissues (Table S1). We also analyze the relationship between: 1) PTENP1 deletion and CDKN2A deletion, as they are located approximately 20cM apart on chromosome 9p (Bennett, 2008); 2) PTENP1 deletion and PTEN deletion, as 9p and 10q losses often coexist in melanoma (Indsto et al., 1998); 3) PTENP1 deletion and BRAF/NRAS mutation, as the combination of mutated BRAF (but not NRAS) and loss of PTEN expression is a common event in human melanoma (Tsao et al., 2004). Our genomic analysis revealed partial deletion of PTENP1 locus in 14.3% of the cell lines tested (Figure 2a). In addition, PTENP1 was found to be deleted in 20.9% of melanoma tissues (8 partial and 1 complete deletion) (Figure 2b). These results illustrate that PTENP1 is under selective pressure to undergo copy number losses (Poliseno et al., 2010). The average PTEN expression level in the 9 samples with either partial or complete deletion of PTENP1 (P5, P9, P11, P12, P20, M1, M9, M14 and M16) was lower than in the remaining 34 samples without any PTENP1 deletion (5.22 ± 2.30 N=9 vs. 20.59 ± 3.98 N=34, p = 0.06), confirming the protective effect of PTENP1 on PTEN levels (Poliseno et al., 2010). Figure. 2 Analysis of CDKN2A, CDKN2B, MTAP, PTENP1, PTEN, BRAF and NRAS status in human melanoma cell lines (a) and tissues (b) The details of the analyses of CDKN2A deletion, PTEN deletion, and BRAF and NRAS mutation in the human melanoma cell lines and tissues are provided in Supplemental Text S2. We observed that the partial or complete deletion of PTENP1 is always concomitant with the partial or complete deletion of CDKN2A in human melanoma cell lines, but not in human melanoma tissues. The “focal” deletion of PTENP1 (5/9 cases) has already been reported in other cancer types, such as breast and colon cancer (Poliseno et al., 2010), but is particularly interesting in melanoma, because it happens in spite of the fact that CDKN2A deletions are the most common alterations (Bennett, 2008). As far as the relationship between the partial or complete deletion of PTENP1 and that of PTEN is concerned, we observed that PTEN is deleted in 7/9 melanoma tissues that show PTENP1 deletion (Figure 2b). This result is supported by previous loss-of-heterozygosity analyses (Herbst et al., 1999) and is in agreement with the observation that 9p and 10q losses often coexist in melanoma (Indsto et al., 1998). The co-deletion of PTEN and PTENP1 can be reconciled with the function of PTENP1 as decoy for PTEN-targeting microRNAs. PTEN deletion, either partial or complete, was found in 23 melanoma tissues. Excluding the cases in which PTEN deletion is complete and therefore PTENP1 deletion cannot affect PTEN levels, on average we observed lower PTEN protein expression in the samples showing both PTEN partial deletion and PTENP1 deletion compared to those showing PTEN partial deletion only, as exemplified by P5 and M5 tissues in Figure 2b. In the context of partial PTEN deletion, PTENP1 deletion may cause a further decrease in PTEN levels, possibly due to less efficient sheltering of PTEN-targeting microRNAs. Of note, PTENP1 deletion may also affect PTEN levels when PTEN is downregulated by mechanisms other than genomic deletion, such as inactivating mutations. The biological effects of PTENP1 deletion are likely to be particularly strong on PTEN because of the intrinsic nature of this haploinsufficient tumor suppressor whose variations of even 20% can have profound consequences on cancer onset and progression (Berger and Pandolfi, 2011). Nonetheless, the concomitant deletion of PTENP1 and PTEN points towards PTEN-independent functions of PTENP1 as well. Two possible explanations can be invoked. First, PTENP1 is likely to function as decoy for additional microRNA families which are not PTEN-targeting. These microRNAs should preferentially bind to the R2 region of the 3’UTR that shows low homology with the corresponding one on PTEN (Figure 1a) and might affect targets that belong to other signaling cascades, so that the concomitant deletion of PTENP1 and PTEN causes the activation of independent pathways that possibly cooperate in tumorigenesis. Second, the PTEN-targeting microRNAs for which PTENP1 acts as decoy have additional targets. Therefore, the deletion of PTENP1 might be advantageous for the tumor because of the decrease of other oncosuppressor genes besides PTEN (Poliseno et al., 2010). Examples of these genes are BIM, p21 and Sprouty2. The pro-apoptotic factor BIM is a miR-17 family-target (Fontana et al., 2008). Mutant BRAF and NRAS induce the MAPK-dependent phosphorylation and consequent proteosomal degradation of BIM in order to provide melanoma cells with resistance to anoikis (Akiyama et al., 2009). The CDK inhibitor p21, another target of miR-17 family (Fontana et al., 2008), is repressed transcriptionally by TBX2 and TBX3 transcription factors which are frequently amplified in melanoma and are downstream effectors of the MAPK pathway (Bennett, 2008). Sprouty2 is a wt BRAF inhibitor that is targeted by miR-21. The downregulation of Sprouty2 observed in melanoma has been hypothesized as an alternative mechanism responsible for the aberrant activation of MAPK pathway besides BRAF mutation (Tsavachidou et al., 2004). Of note, the three genes described above share a common feature as antagonists of the MAPK pathway. Since the PI3K/AKT and the MAPK pathways have been shown to cooperate in melanomagenesis both in vitro and in mouse models (Dankort et al., 2009; Meier et al., 2007), it is tempting to hypothesize that, in the context of PTEN deletion, the deletion of PTENP1 might be an alternative mechanism evolved by the tumor to activate the MAPK pathway. In this respect, it is worth noticing that most of the cases showing both PTENP1 and PTEN deletion (71.4%) harbor wt BRAF. In conclusion, our data indicate that the recently identified tumor suppressor gene PTENP1 undergoes genomic deletion in human melanoma. Our data also suggest that PTENP1 deletion might be advantageous for the tumor not only because of its PTEN-related function, but also for PTEN-unrelated ones.

The novel gamma secretase inhibitor RO4929097 reduces the tumor initiating potential of melanoma.

Several reports have demonstrated a role for aberrant NOTCH signaling in melanoma genesis and progression, prompting us to explore if targeting this pathway is a valid therapeutic approach against melanoma. We targeted NOTCH signaling using RO4929097, a novel inhibitor of gamma secretase, which is a key component of the enzymatic complex that cleaves and activates NOTCH. The effects of RO4929097 on the oncogenic and stem cell properties of a panel of melanoma cell lines were tested both in vitro and in vivo, using xenograft models. In human primary melanoma cell lines, RO4929097 decreased the levels of NOTCH transcriptional target HES1. This was accompanied by reduced proliferation and impaired ability to form colonies in soft agar and to organize in tridimensional spheres. Moreover, RO4929097 affected the growth of human primary melanoma xenograft in NOD/SCID/IL2gammaR-/- mice and inhibited subsequent tumor formation in a serial xenotransplantation model, suggesting that inhibition of NOTCH signaling suppresses the tumor initiating potential of melanoma cells. In addition, RO4929097 decreased tumor volume and blocked the invasive growth pattern of metastatic melanoma cell lines in vivo. Finally, increased gene expression of NOTCH signaling components correlated with shorter post recurrence survival in metastatic melanoma cases. Our data support NOTCH inhibition as a promising therapeutic strategy against melanoma.

Histology-Specific MicroRNA Alterations in Melanoma

We examined the microRNA signature that distinguishes the most common melanoma histological subtypes, superficial spreading melanoma (SSM) and nodular melanoma (NM). We also investigated the mechanisms underlying the differential expression of histology-specific microRNAs. MicroRNA array performed on a training cohort of 82 primary melanoma tumors (26 SSM, 56 NM), and nine congenital nevi (CN) revealed 134 microRNAs differentially expressed between SSM and NM (P<0.05). Out of 134 microRNAs, 126 remained significant after controlling for thickness and 31 were expressed at a lower level in SSM compared with both NM and CN. For seven microRNAs (let-7g, miR-15a, miR-16, miR-138, miR-181a, miR-191, and miR-933), the downregulation was associated with selective genomic loss in SSM cell lines and primary tumors, but not in NM cell lines and primary tumors. The lower expression level of six out of seven microRNAs in SSM compared with NM was confirmed by real-time PCR on a subset of cases in the training cohort and validated in an independent cohort of 97 melanoma cases (38 SSM, 59 NM). Our data support a molecular classification in which SSM and NM are two molecularly distinct phenotypes. Therapeutic strategies that take into account subtype-specific alterations might improve the outcome of melanoma patients.

Patient perspectives on medical photography in dermatology.

BACKGROUND Clinical photography enhances medical care, research, and teaching. Empirical data are needed to guide best practices regarding dermatologic photography. OBJECTIVE To investigate patient opinion about clinical photography and identify demographic factors that influence these opinions. METHODS AND MATERIALS Four hundred patients representing a broad range of ages, self-identified ethnic/racial groups, and socioeconomic levels were recruited from 4 dermatology settings in New York City. Patients were administered a survey about perceptions of photography, willingness to allow photographs to be used in a variety of settings, preferences for photographer and photographic equipment, and methods of consent. RESULTS Eighty-eight percent of patients agreed that photography enhanced their quality of care. Most patients would allow their photographs to be used for medical, teaching, and research purposes with significantly more acceptance when patients were not identifiable. Patients preferred photographs taken by a physician rather than a nurse or student, photographers of the same gender, clinic-owned cameras to personal cameras or cell phones, and written consent to verbal consent. There were significant racial/ethnicity and age-related variations in responses, with white and older patients being more permissive than other groups. CONCLUSION We use the results of this study to recommend best practices for photography in dermatology.

Microneedling: A Comprehensive Review.

BACKGROUND Microneedling is a minimally invasive procedure that uses fine needles to puncture the epidermis. The microwounds created stimulate the release of growth factors and induce collagen production. The epidermis remains relatively intact, therefore helping to limit adverse events. The indications for microneedling therapy have grown significantly, and it is becoming a more widely used treatment in dermatology. OBJECTIVE A comprehensive review of microneedling in human subjects and its applications in dermatology. METHODS AND MATERIALS A search was performed using PubMed/MEDLINE and Science Direct databases. Search terms included “microneedling,” “needling,” and “percutaneous collagen induction.” All available studies involving human subjects were included in the discussion, with priority given to prospective, randomized trials. RESULTS Studies demonstrate microneedling efficacy and safety for the treatment of scars, acne, melasma, photodamage, skin rejuvenation, hyperhidrosis and alopecia and for facilitation of transdermal drug delivery. While permanent adverse events are uncommon, transient erythema and postinflammatory hyperpigmentation are more commonly reported. CONCLUSION Microneedling appears to be an overall effective and safe therapeutic option for numerous dermatologic conditions. Larger and more randomized controlled trials are needed to provide greater data on the use of microneedling for different dermatologic conditions in different skin types.

Safety of a picosecond laser with diffractive lens array (DLA) in the treatment of Fitzpatrick skin types IV to VI: A retrospective review

BACKGROUND Laser therapy in patients with skin of color is associated with an increased rate of complications. The 755-nm picosecond laser with the diffractive lens array (DLA) has been used for the treatment of scars, striae, and rejuvenation. By delivering high energy to focused areas, the DLA minimizes complications. OBJECTIVE This study explores the adverse events associated with treatment with the 755-nm picosecond laser with DLA in individuals with Fitzpatrick skin type IV to VI. METHOD A retrospective chart review of patients treated with the 755-nm picosecond laser with DLA with a standardized spot size of 6 mm, fluence of 0.71 J/cm(2), and pulse width of 750 to 850 picoseconds was performed. Standard clinical photographs were obtained before treatment and at follow-up. Treatment sites were assessed for dyspigmentation, erythema, edema, and herpetic lesions. RESULTS A total of 56 patients with Fitzpatrick skin type IV to VI, atrophic and hypertrophic scars, and pigmented lesions or striae were included. Ten patients (17.9%) were lost to follow-up. Transient adverse events, most commonly erythema and hyperpigmentation, were reported after therapy; these resolved in all cases. LIMITATIONS Retrospective design is a limitation. CONCLUSION The 755-nm picosecond laser with the DLA device may be a safe therapeutic alternative for unwanted scars, pigmented lesions, and striae in patients with skin of color.

Continuing medical educationSarcoidosis: A comprehensive review and update for the dermatologist: Part II. Extracutaneous disease

Sarcoidosis is a multisystemic, granulomatous disease with protean manifestations and variable prognosis. Because the skin can be the only organ in which the disease is recognized, dermatologists may be responsible for the care of sarcoidosis patients. Therefore, dermatologists should be cognizant of the diseases extracutaneous manifestations to assure appropriate evaluation and treatment. Part II of this review describes the diagnostic approach and management of the extracutaneous manifestations of sarcoidosis.

Reply to: Monogenic early-onset sarcoidosis is no longer a variant of “idiopathic” sarcoidosis

REFERENCES 1. Haimovic A, Sanchez M, Judson MA, Prystowsky S. Sarcoidosis: a comprehensive review and update for the dermatologist: part I. Cutaneous disease. J Am Acad Dermatol 2012;66:699.e1-18. 2. Kanazawa N, Okafuji I, Kambe N, Nishikomori R, Nakata-Hizume M, Nagai S, et al. Early-onset sarcoidosis and CARD15 mutations with constitutive nuclear factor-kappaB activation: common genetic etiology with Blau syndrome. Blood 2005;105:1195-7.

Self-reported Patient Motivations for Seeking Cosmetic Procedures

Importance Despite the growing popularity of cosmetic procedures, the sociocultural and quality-of-life factors that motivate patients to undergo such procedures are not well understood. Objective To estimate the relative importance of factors that motivate patients to seek minimally invasive cosmetic procedures. Design, Setting, and Participants This prospective, multicenter observational study was performed at 2 academic and 11 private dermatology practice sites that represented all US geographic regions. Adult patients presenting for cosmetic consultation or treatment from December 4, 2016, through August 9, 2017, were eligible for participation. Exposures Participants completed a survey instrument based on a recently developed subjective framework of motivations and a demographic questionnaire. Main Outcomes and Measures Primary outcomes were the self-reported most common motivations in each quality-of-life category. Secondary outcomes were other frequently reported motivations and those associated with specific procedures. Results Of 529 eligible patients, 511 agreed to participate, were enrolled, and completed the survey. Typical respondents were female (440 [86.1%]), 45 years or older (286 [56.0%]), white (386 [75.5%]), and college educated (469 [91.8%]) and had previously received at least 2 cosmetic procedures (270 [52.8%]). Apart from motivations pertaining to aesthetic appearance, including the desire for beautiful skin and a youthful, attractive appearance, motives related to physical health, such as preventing worsening of condition or symptoms (253 of 475 [53.3%]), and psychosocial well-being, such as the desire to feel happier and more confident or improve total quality of life (314 of 467 [67.2%]), treat oneself or celebrate (284 of 463 [61.3%]), and look good professionally (261 of 476 [54.8%]) were commonly reported. Motivations related to cost and convenience were rated as less important (68 of 483 [14.1%]). Most motivations were internally generated, designed to please the patients and not others, with patients making the decision to undergo cosmetic procedures themselves and spouses seldom being influential. Patients younger than 45 years were more likely to undertake procedures to prevent aging (54 of 212 [25.5%] vs 42 of 286 [14.7%] among patients ≥45 years; P < .001). Patients seeking certain procedures, such as body contouring (19 of 22 [86.4%]), acne scar treatment (36 of 42 [85.7%]), and tattoo removal (8 of 11 [72.7%]), were more likely to report psychological and emotional motivations. Conclusions and Relevance This initial prospective, multicenter study comprehensively assessed why patients seek minimally invasive cosmetic procedures. Common reasons included emotional, psychological, and practical motivations in addition to the desire to enhance physical appearance. Differences relative to patient age and procedures sought may need further exploration.