Induced pluripotent stem cell-derived neuronal cells from a sporadic Alzheimer's disease donor as a model for investigating AD-associated gene regulatory networks.

BACKGROUND: Alzheimer's disease (AD) is a complex, irreversible neurodegenerative disorder. At present there are neither reliable markers to diagnose AD at an early stage nor therapy. To investigate underlying disease mechanisms, induced pluripotent stem cells (iPSCs) allow the generation of patient-derived neuronal cells in a dish. RESULTS: In this study, employing iPS technology, we derived and characterized iPSCs from dermal fibroblasts of an 82-year-old female patient affected by sporadic AD. The AD-iPSCs were differentiated into neuronal cells, in order to generate disease-specific protein association networks modeling the molecular pathology on the transcriptome level of AD, to analyse the reflection of the disease phenotype in gene expression in AD-iPS neuronal cells, in particular in the ubiquitin-proteasome system (UPS), and to address expression of typical AD proteins. We detected the expression of p-tau and GSK3B, a physiological kinase of tau, in neuronal cells derived from AD-iPSCs. Treatment of neuronal cells differentiated from AD-iPSCs with an inhibitor of γ-secretase resulted in the down-regulation of p-tau. Transcriptome analysis of AD-iPS derived neuronal cells revealed significant changes in the expression of genes associated with AD and with the constitutive as well as the inducible subunits of the proteasome complex. The neuronal cells expressed numerous genes associated with sub-regions within the brain thus suggesting the usefulness of our in-vitro model. Moreover, an AD-related protein interaction network composed of APP and GSK3B among others could be generated using neuronal cells differentiated from two AD-iPS cell lines. CONCLUSIONS: Our study demonstrates how an iPSC-based model system could represent (i) a tool to study the underlying molecular basis of sporadic AD, (ii) a platform for drug screening and toxicology studies which might unveil novel therapeutic avenues for this debilitating neuronal disorder.

Acne tarda and male-pattern baldness unmasking primary ovarian insufficiency: a case and review.

A 30-year-old woman presented with recurrent acne lesions and progressing male-pattern baldness. Furthermore, she reported amenorrhea, weight loss, mucosal xerosis and dyspareunia since discontinuation of hormonal contraception 6 months earlier in order to conceive. Acne tarda and androgenetic alopecia of female pattern were diagnosed. Hormonal and immunologic serological and ultrasound examinations revealed an autoimmune hypergonadotropic primary ovarian insufficiency (POI) with no ovarian cysts but ovarian fibrosis with marked reduced follicle pool. Immediate ovarian stimulation and in vitro fertilization led to pregnancy and the patient gave birth to a healthy child. Though presenting with clinical findings similar to menopause, 50% of patients with POI exhibit varying and unpredictable ovarian function, and only 5-10% are able to accomplish pregnancy. Genetic disorders affect the X chromosome. In 14-30% of cases POI has been associated with autoimmunity. POI may occur after discontinuation of hormonal contraception, like in our case.

[Skin aging exposome]. / Hautalterungsexposom.

The skin is a barrier organ and thus exposed to environmental factors from birth, which essentially determine skin aging. In order to describe and understand this complex process exactly, we applied the concept of the "exposome" to the environmentally induced skin aging process. In this review, we summarize current knowledge on the skin aging exposome. In this context, we characterize the most important exposomal factors, address their relative importance for skin aging and also the relevance of their mutual interactions. Finally, we discuss the clinical consequences resulting from this concept for an effective prevention of skin aging.

Clinical and laboratory skin biomarkers of organ-specific diseases.

Intense research during the last few years has elucidated a number of mechanisms of the complex human aging process. Among the body organs, skin has a distinguished role in aging perception both because it constitutes the organ on which the first signs of aging are mostly visible and because skin is recognized as an indicator of human health. Analogous to all other organs, skin undergoes multiple morphological and functional changes with age affected by extrinsic and intrinsic factors. Evaluation of skin morphology as well as of advanced glycation end products, dermal collagen content and alterations of the Wnt signaling pathway may predict - among others - the aging status of the cardiovascular system, the brain and the bones. Increasing knowledge on the association of certain clinical and laboratory characteristics of extrinsic and intrinsic skin aging with organ-specific diseases opens new possibilities for the establishment of clinical and laboratory skin biomarkers for systemic human aging and the course of several internal, organ-specific diseases.

When the skin is in the center of interest: An aging issue.

The skin represents the first bearer of marks of time as well as an easily accessible model for the assessment and determination of the involved molecular mechanisms. The deterioration of important skin functions due to intrinsic and extrinsic aging leads to clinical manifestations, which mirror several internal age-associated diseases, such as neurodegenerative, cardiovascular, skeletal, and endocrine/metabolic skin diseases. Current molecular data indicate that skin aging, especially intrinsic aging, mirrors age-related deficiencies in the entire human body. These data and the development of new biologic technologies highlight the importance of the skin in aging research and should enable future interdisciplinary projects on internal diseases, which could barely have been performed until recently due mainly to the lack of respective tissue.

Aesthetic aspects of skin aging, prevention, and local treatment.

Skin aging is a complex biologic process influenced by a combination of intrinsic and extrinsic factors. Aging skin shows wrinkles, uneven tone, loss of elasticity, and thinning. Skin health is considered one of the principal factors representing overall well-being and the perception of health in humans; therefore, anti-aging strategies to combat aging signs and dysfunction have been developed over the last decades. Understanding the mechanism behind skin aging is required for elucidation of the mechanism of action and, hence, the potential benefits of the claimed anti-aging products. In this review, preventive measurements, cosmetologic strategies, and photoprotection (systemic antioxidants, ultraviolet and filters), as well as the mechanisms of action and the effectiveness of topical pharmaceutical agents, such as antioxidants (vitamins, polyphenols, and flavonoids) and cell regulators (retinols, peptides, hormones, and botanicals), are presented.

Relaxin 2 is functional at the ocular surface and promotes corneal wound healing.

PURPOSE: We aimed to determine if the insulin-like peptide hormone relaxin 2 (RLN2) is expressed at the ocular surface and in tears and if RLN2 influences wound healing at the ocular surface, which is associated with extracellular matrix (ECM) remodeling. METHODS: We analyzed transcript levels of human RLN2 and its cognate relaxin-like receptors RXFP1 and RXFP2 in tissues of the ocular surface, lacrimal apparatus, and human corneal (HCE), conjunctival (HCjE) and sebaceous (SC) cell lines. We analyzed effects of human RLN2 on cell proliferation and migration and quantified mRNA expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in HCE, HCjE, and SC. Using an alkali-induced corneal wounding model, we analyzed the wound healing rate in C57BL/6 mice eyes after topically applied RLN2. RESULTS: The presence of RLN2, RXFP1, and RXFP2 transcripts was detected in lacrimal gland, eyelid, conjunctiva, cornea, primary corneal fibroblasts, nasolacrimal ducts, and all three cell lines. ELISA revealed RLN2 protein in all ocular surface tissues analyzed and in human tears. Stimulation of HCE, HCjE, and SC with RLN2 significantly increased cell proliferation and migration. Relative mRNA expression levels of MMP2, MMP9, TIMP1, and TIMP2 were significantly influenced by RLN2 in all three cell lines at different time points studied. The local application of RLN2 onto denuded corneal surface resulted in significantly elevated corneal wound healing. CONCLUSIONS: Our data support a novel role for the RLN2 ligand-receptor system at the ocular surface and in the lacrimal apparatus as a potential future therapeutic during wound healing at the ocular surface.

Mitochondrial-associated cell death mechanisms are reset to an embryonic-like state in aged donor-derived iPS cells harboring chromosomal aberrations.

Somatic cells reprogrammed into induced pluripotent stem cells (iPSCs) acquire features of human embryonic stem cells (hESCs) and thus represent a promising source for cellular therapy of debilitating diseases, such as age-related disorders. However, reprogrammed cell lines have been found to harbor various genomic alterations. In addition, we recently discovered that the mitochondrial DNA of human fibroblasts also undergoes random mutational events upon reprogramming. Aged somatic cells might possess high susceptibility to nuclear and mitochondrial genome instability. Hence, concerns over the oncogenic potential of reprogrammed cells due to the lack of genomic integrity may hinder the applicability of iPSC-based therapies for age-associated conditions. Here, we investigated whether aged reprogrammed cells harboring chromosomal abnormalities show resistance to apoptotic cell death or mitochondrial-associated oxidative stress, both hallmarks of cancer transformation. Four iPSC lines were generated from dermal fibroblasts derived from an 84-year-old woman, representing the oldest human donor so far reprogrammed to pluripotency. Despite the presence of karyotype aberrations, all aged-iPSCs were able to differentiate into neurons, re-establish telomerase activity, and reconfigure mitochondrial ultra-structure and functionality to a hESC-like state. Importantly, aged-iPSCs exhibited high sensitivity to drug-induced apoptosis and low levels of oxidative stress and DNA damage, in a similar fashion as iPSCs derived from young donors and hESCs. Thus, the occurrence of chromosomal abnormalities within aged reprogrammed cells might not be sufficient to over-ride the cellular surveillance machinery and induce malignant transformation through the alteration of mitochondrial-associated cell death. Taken together, we unveiled that cellular reprogramming is capable of reversing aging-related features in somatic cells from a very old subject, despite the presence of genomic alterations. Nevertheless, we believe it will be essential to develop reprogramming protocols capable of safeguarding the integrity of the genome of aged somatic cells, before employing iPSC-based therapy for age-associated disorders.