Network-based prioritization and validation of regulators of vascular smooth muscle cell proliferation in disease.

Aberrant vascular smooth muscle cell (VSMC) homeostasis and proliferation characterize vascular diseases causing heart attack and stroke. Here we elucidate molecular determinants governing VSMC proliferation by reconstructing gene regulatory networks from single-cell transcriptomics and epigenetic profiling. We detect widespread activation of enhancers at disease-relevant loci in proliferation-predisposed VSMCs. We compared gene regulatory network rewiring between injury-responsive and nonresponsive VSMCs, which suggested shared transcription factors but differing target loci between VSMC states. Through in silico perturbation analysis, we identified and prioritized previously unrecognized regulators of proliferation, including RUNX1 and TIMP1. Moreover, we showed that the pioneer transcription factor RUNX1 increased VSMC responsiveness and that TIMP1 feeds back to promote VSMC proliferation through CD74-mediated STAT3 signaling. Both RUNX1 and the TIMP1-CD74 axis were expressed in human VSMCs, showing low levels in normal arteries and increased expression in disease, suggesting clinical relevance and potential as vascular disease targets.

TIMP-1 is a novel ligand of Amyloid Precursor Protein and triggers a proinflammatory phenotype in human monocytes.

The emerging cytokine tissue inhibitor of metalloproteinases-1 (TIMP-1) correlates with the progression of inflammatory diseases, including cancer. However, the effects of TIMP-1 on immune cell activation and underlying molecular mechanisms are largely unknown. Unbiased ligand-receptor-capture-screening revealed TIMP-1-interaction with Amyloid Precursor Protein (APP) family members, namely APP and Amyloid Precursor-like Protein-2 (APLP2), which was confirmed by pull-down assays and confocal microscopy. We found that TIMP-1 triggered glucose uptake and proinflammatory cytokine expression in human monocytes. In cancer patients, TIMP-1 expression positively correlated with proinflammatory cytokine expression and processes associated with monocyte activation. In pancreatic cancer, TIMP-1 plasma levels correlated with the monocyte activation marker sCD163, and the combined use of both clinically accessible plasma proteins served as a powerful prognostic indicator. Mechanistically, TIMP-1 triggered monocyte activation by its C-terminal domain and via APP as demonstrated by in vitro interference, in silico docking, and the employment of recombinant TIMP-1 variants. Identification of TIMP-1 as a trigger of monocyte activation opens new therapeutic perspectives for inflammatory diseases.

The effect of complementary music intervention on the patients' quality of life after septoplasty and rhinoplasty.

PURPOSE: Quality of life (QoL) assessment has emerged as an important evaluation tool for therapeutic treatments. The positive impact of complementary music interventions on QoL has been demonstrated in the literature, particularly in chronic and malignant diseases. However, its benefits during the perioperative period in head and neck patients have not been investigated thus far. METHODS: Head and neck patients undergoing septoplasty and rhinoplasty were prospectively randomized and consecutively included in the trial. Passive music intervention (60 min per day) was applied to the intervention group. QoL was assessed using the Nasal Obstruction Symptom Evaluation (NOSE) questionnaire and the Functional Rhinoplasty Outcome Inventory 17 (FROI-17) questionnaire at three visits during the postoperative phase. Pain was measured using a visual analogue scale. RESULTS: Forty-four patients were enrolled in the study. The NOSE score between the control group and the intervention group in the septoplasty arm differed significantly at visit #2 (p < 0.001) and visit #3 (p < 0.015). For the rhinoplasty study arm, significant differences in the FROI-17 score were also found at visit #2 and visit #3 (p = 0.04). CONCLUSION: Complementary music interventions can considerably improve patients' QoL during the postoperative period. Furthermore, passive music interventions may be easily implemented in clinical practice as an additional cost-effective treatment with ubiquitous availability.

Long-Term Results of a Multimodal Treatment Regimen in Patients With Auricular Keloids.

Objectives: The treatment of auricular keloids is challenging, as they tend to recur; further, the treatment may impact quality of life and implies cosmetic and functional impairment for each patient. There is no standardized therapeutic concept established, and the literature is lacking long-term results of available treatment modalities. Methods: Patients suffering from auricular keloids were included in the study. All patients had undergone surgical resection, intralesional injection of triamcinolone acetonide (TAC), and the application of an individual pressure splint. Quality of life (QoL) was assessed using the keloid intervention benefit inventory 21 (KIBI-21). Further analysis was carried out for patients without (group 1) and with (group 2) recurrence of the keloid. Results: In total, 50 keloids with a mean follow-up period of 59 months (range 6-137 months) could be analyzed. In nine cases (18%), a keloid recurrence was found during the observation period. The assessment of QoL differed significantly between study groups at P = 0.04, as well as for the subcategories General Health (GH) and Physical Health (PH). No differences were found for the categories Social Impact (SI) and Self-Esteem (SE). Conclusions: The multimodal subsequent treatment regimen consisting of surgical resection, intralesional TAC injection, and the application of an individual magnetic pressure splint shows good results concerning long-term recurrence rates. The treatment method shows positive effects on the QoL, especially in the measured categories GH and PH.

An in vitro study on the effect of bevacizumab on endothelial cell proliferation and VEGF concentration level in patients with hereditary hemorrhagic telangiectasia.

Previous studies have demonstrated that vascular endothelial growth factor (VEGF) is upregulated in patients with hereditary hemorrhagic telangiectasia (HHT). The use of Bevacizumab as an anti-angiogenic treatment agent seems promising. The purpose of the present in vitro study was to determine the efficacy and potential toxicity levels of bevacizumab on cell proliferation and VEGF concentrations in endothelial cells of HHT patients. In this in vitro study, endothelial cells from patients with HHT and HUVECs (control) were incubated with different concentration levels of bevacizumab (2, 4, 6, 8 or 10 mg/ml). After 24, 48 or 72 h, the cell proliferation was assessed by Alamar Blue® Assay and the VEGF levels in the cell culture supernatants were measured by VEGF-ELISA. All endothelial cells incubated with bevacizumab showed an initial decrease in cell proliferation. Cell proliferation recovered within 72 h in cell cultures incubated with concentration levels of up to 4 mg/ml bevacizumab, whereas those incubated with higher concentration levels showed a continuous decline in cell proliferation. VEGF expression decreased after 24 h in cell cultures incubated with bevacizumab concentration levels of 2 and 4 mg/ml but increased again after 48 h. Cell cultures incubated with bevacizumab concentration levels of 10 mg/ml showed a constant decline in VEGF expression without any tendency for recovery. Translating these results into daily clinical practice, the present study suggests that the intranasal submucosal injection of bevacizumab in HHT patients should not exceed a concentration level of 4 mg/ml. Overall, higher bevacizumab concentration levels not only reduce VEGF expression but pose a higher risk of toxic effects on endothelial cells as they jeopardize cell proliferation.

TIMP1 expression underlies sex disparity in liver metastasis and survival in pancreatic cancer.

Sex disparity in cancer is so far inadequately considered, and components of its basis are rather unknown. We reveal that male versus female pancreatic cancer (PC) patients and mice show shortened survival, more frequent liver metastasis, and elevated hepatic metastasis-promoting gene expression. Tissue inhibitor of metalloproteinases 1 (TIMP1) was the secreted factor with the strongest male-biased expression in patient-derived pancreatic tumors. Male-specific up-regulation of systemic TIMP1 was demonstrated in PC mouse models and patients. Using TIMP1-competent and TIMP1-deficient PC mouse models, we established a causal role of TIMP1 in determining shortened survival and increased liver metastasis in males. Observing TIMP1 expression as a risk parameter in males led to identification of a subpopulation exhibiting increased TIMP1 levels (T1HI males) in both primary tumors and blood. T1HI males showed increased risk for liver metastasis development not only in PC but also in colorectal cancer and melanoma. This study reveals a lifestyle-independent sex disparity in liver metastasis and may open new avenues toward precision medicine.

Identification of invariant chain CD74 as a functional receptor of tissue inhibitor of metalloproteinases-1 (TIMP-1).

Multifunctionality of tissue inhibitor of metalloproteinases-1 (TIMP-1) comprising antiproteolytic as well as cytokinic activity has been attributed to its N-terminal and C-terminal domains, respectively. The molecular basis of the emerging proinflammatory cytokinic activity of TIMP-1 is still not completely understood. The cytokine receptor invariant chain (CD74) is involved in many inflammation-associated diseases and is highly expressed by immune cells. CD74 triggers zeta chain-associated protein kinase-70 (ZAP-70) signaling-associated activation upon interaction with its only known ligand, the macrophage migration inhibitory factor. Here, we demonstrate TIMP-1-CD74 interaction by coimmunoprecipitation and confocal microscopy in cells engineered to overexpress CD74. In silico docking in HADDOCK predicted regions of the N-terminal domain of TIMP-1 (N-TIMP-1) to interact with CD74. This was experimentally confirmed by confocal microscopy demonstrating that recombinant N-TIMP-1 lacking the entire C-terminal domain was sufficient to bind CD74. Interaction of TIMP-1 with endogenously expressed CD74 was demonstrated in the Namalwa B lymphoma cell line by dot blot binding assays as well as confocal microscopy. Functionally, we demonstrated that TIMP-1-CD74 interaction triggered intracellular ZAP-70 activation. N-TIMP-1 was sufficient to induce ZAP-70 activation and interference with the cytokine-binding site of CD74 using a synthetic peptide-abrogated TIMP-1-mediated ZAP-70 activation. Altogether, we here identified CD74 as a receptor and mediator of cytokinic TIMP-1 activity and revealed TIMP-1 as moonlighting protein harboring both cytokinic and antiproteolytic activity within its N-terminal domain. Recognition of this functional TIMP-1-CD74 interaction may shed new light on clinical attempts to therapeutically target ligand-induced CD74 activity in cancer and other inflammatory diseases.

TIMP1 Triggers Neutrophil Extracellular Trap Formation in Pancreatic Cancer.

Tumor-derived protein tissue inhibitor of metalloproteinases-1 (TIMP1) correlates with poor prognosis in many cancers, including highly lethal pancreatic ductal adenocarcinoma (PDAC). The noncanonical signaling activity of TIMP1 is emerging as one basis for its contribution to cancer progression. However, TIMP1-triggered progression-related biological processes are largely unknown. Formation of neutrophil extracellular traps (NET) in the tumor microenvironment is known to drive progression of PDAC, but factors or molecular mechanisms initiating NET formation in PDAC remain elusive. In this study, gene-set enrichment analysis of a human PDAC proteome dataset revealed that TIMP1 protein expression most prominently correlates with neutrophil activation in patient-derived tumor tissues. TIMP1 directly triggered formation of NETs in primary human neutrophils, which was dependent on the interaction of TIMP1 with its receptor CD63 and subsequent ERK signaling. In genetically engineered PDAC-bearing mice, TIMP1 significantly contributed to NET formation in tumors, and abrogation of TIMP1 or NETs prolonged survival. In patient-derived PDAC tumors, NETs predominantly colocalized with areas of elevated TIMP1 expression. Furthermore, TIMP1 plasma levels correlated with DNA-bound myeloperoxidase, a NET marker, in the blood of patients with PDAC. A combination of plasma levels of TIMP1 and NETs with the clinically established marker CA19-9 allowed improved identification of prognostically distinct PDAC patient subgroups. These observations may have a broader impact, because elevated systemic levels of TIMP1 are associated with the progression of a wide range of neutrophil-involved inflammatory diseases. SIGNIFICANCE: These findings highlight the prognostic relevance of TIMP1 and neutrophil extracellular traps in highly lethal pancreatic cancer, where a noncanonical TIMP1/CD63/ERK signaling axis induces NET formation. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/81/13/3568/F1.large.jpg.

The Keloid Intervention Benefit Inventory 21: A New Assessment Tool for the Quality of Life of Patients with Auricular Keloids.

The assessment of the quality of life (QoL) of patients with chronic diseases before and after medical interventions has gained increasing importance in recent decades. Particularly for patients with visible keloid scars in the head and neck region, standardized measurement tools are either absent or have been shown to be insufficient. The aim of the present study was to create a new standardized questionnaire that is specific to auricular keloid patients and reflects their clinical symptoms and QoL. The Keloid Intervention Benefit Inventory 21 (KIBI-21) questionnaire was developed in two stages. First, a group of experts identified a pool of 26 questions and modified and supplemented the items through a comparison with existing QoL assessments so that they related to keloid-specific clinical symptoms and the QoL of patients with auricular keloids before and after a medical intervention. This questionnaire was distributed to 27 outpatients who had undergone medical interventions for visible auricular keloids. Second, a sequential statistical analysis was conducted. This included a single-item assessment and reduction, analysis for internal consistency, construct validity, and divergence validity as well as a factor analysis. The analyses were performed for the entire questionnaire and for the items in the subcategories General Health, Physical Symptoms, Self-Esteem, and Social Impact. The final version of this newly validated and standardized KIBI questionnaire consisted of 21 items, of which each item was assigned to only one subscale. The questionnaire showed a Cronbach's α of 0.84 with a good internal consistency. In the item correlation validity, strong associations were found in all subscales, except for the Social Impact Subscale. The keloid-specific QoL questionnaire KIBI-21 proved to be a reliable and reproducible instrument to assess the QoL and clinical symptoms in patients suffering from auricular keloids before and after a medical treatment.

Influence of craniocervical and craniomandibular dysfunction to nonrestorative sleep and sleep disorders.

Objective: Nonrestorative sleep and sleep disorders are commonly reported in patients suffering from craniomandibular (CMD) and craniocervical dysfunctions (CCD). This study aimed to investigate polysomnographic characteristics and the reduction of pain before and after treatment in these patients.Methods: Seventy-four patients with sleep disorders and evident CMD and CCD were included. Manual therapy and an Aqualizer® were used in the therapeutic group. Polysomnographic measurements were conducted pre- and post-therapy.Results: The number of sleep stage alterations and the sleep stage index differed significantly between pre- and post-therapeutic measurements. Between both groups, these parameters were significantly different, as well (p = .001 and p = .012). The subjective perception of sleep quality improved in 81.6% post-therapy (p < .001).Discussion: Manual therapy and the application of an Aqualizer® may improve sleep quality in patients suffering from CMD and CCD. Pain may not be the main cause for the sleep disorders in CMD and CCD.

[Options for reconstruction of nasal defects]. / Möglichkeiten der Defektrekonstruktion im Nasenbereich.

As far as aesthetic and functional aspects are concerned, local as well as regional flaps are mainly recommended for facial plastic surgery. A careful preoperative planning is essential. Adequate knowledge of various surgical options of reconstruction as well as their implementation into clinical practice are of utmost importance. The surgical procedure is selected on the basis of the patients' demands and the properties of the surgical site to which the selected technique must be adapted. The current review presents frequently used and recommendable methods for the reconstruction of nasal defects.

Functional disparities within the TIMP family in cancer: hints from molecular divergence.

The members of the tissue inhibitor of metalloproteinase (TIMP) family (TIMP-1, 2, 3, 4) are prominently appreciated as natural inhibitors of cancer-promoting metalloproteinases. However, clinical and recent functional studies indicate that some of them correlate with bad prognosis and contribute to the progression of cancer and metastasis, pointing towards mechanisms beyond inhibition of cancer-promoting proteases. Indeed, it is increasingly recognized that TIMPs are multi-functional proteins mediating a variety of cellular effects including direct cell signaling. Our aim was to provide comprehensive information towards a better appreciation and understanding of the biological heterogeneity and complexity of the TIMPs in cancer. Comparison of all four members revealed distinct cancer-associated expression patterns and distinct prognostic impact including a clear correlation of TIMP-1 with bad prognosis for almost all cancer types. For the first time, we present the interactomes of all TIMPs regarding overlapping and non-overlapping interaction partners. Interestingly, the overlap was maximal for metalloproteinases (e.g., matrix metalloproteinase 1, 2, 3, 9) and decreased for non-protease molecules, especially cell surface receptors (e.g., CD63, overlapping only for TIMP-1 and 4; IGF-1R unique for TIMP-2; VEGFR2 unique for TIMP-3). Finally, we attempted to identify and summarize experimental evidence for common and unique structural traits of the four TIMPs on the basis of amino acid sequence and protein folding, which account for functional disparities. Altogether, the four TIMPs have to be appreciated as molecules with commonalities, but, more importantly, functional disparities, which need to be investigated further in the future, since those determine their distinct roles in cancer and metastasis.

[Facial skin tumors and plastic reconstruction: part 2]. / Faziale Hauttumore: Resektion und plastische Defektrekonstruktion ­ Teil 2.

Reconstruction of defects resulting from the resection of skin tumors in the face depends on patient-specific factors such as prior radiation therapy or prior surgery, the patient´s individual expectations and the skills of the surgeon. Local flaps are favorable in several respects, however they require a detailed plan and experience when being executed. The following article provides an overview of the most frequently used local flaps which have a high reliability for reconstruction. Defects in specific anatomical areas of the nose, the ears, the cheek and the forehead are described and analysed. The authors suggest possible surgical reconstruction strategies which are highlighted by clinical cases.

[Facial skin tumors and plastic reconstruction: part 1]. / Faziale Hauttumore: Resektion und plastische Defektrekonstruktion ­ Teil 1.

Skin tumors are becoming more and more frequent due to an increased exposition to UV-radiation and the growing age of the population. At the same time expectations with regard to function and aesthetics are growing. An in-total resection of the tumor with clear margins is the first step in surgical treatment of such patients. Reconstruction of the resulting defects depends on patient-specific factors such as prior radiation therapy or prior surgery, the patient´s individual expectations and the surgeon´s skills. Local flaps are favorable in several respects, however they require a detailed plan and experience when being executed. The following article provides an overview of the most frequent facial skin tumors and the secure margins they require. Furthermore, it gives a general overview of the design of local flaps and grafts.