Assoziation von Genvarianten mit Bezug zur Hautpigmentierung und Vitamin D Status

Hintergrund: In der kaukasischen Bevölkerung besteht häufig ein Vitamin D Mangel. Dieser ist bei vielen Erkrankungen, wie zum Beispiel diversen Krebserkrankungen, Autoimmunkrankheiten, Infektionen und kardiovaskulären Krankheiten mit erhöhten Inzidenzen sowie einem ungünstigeren klinischen Verlauf assoziiert. Vorherige Arbeiten haben bereits einige der Faktoren, welche den Vitamin D Status beeinflussen, identifiziert, jedoch liegen bisher sehr wenige Untersuchungen zur Relevanz von genetischen Variationen für die 25(OH)D Serumkonzentration vor. Diese Arbeit analysiert die Hypothese, dass Varianten von Genen (SNPs), welche mit der Hautpigmentierung im Zusammenhang stehen, den 25(OH)D Serumspiegel einer Person beeinflussen. Patienten und Methoden: In einer Kohorte der LURIC Studie (n=2974) wurde die mögliche Assoziation von 25(OH)D Serumkonzentration (Median der Kohorte: 15,5 ng/ml) und SNPs (n=244) in der Nähe von bzw. in Genen, welche die Hautpigmentierung beeinflussen (n=15) untersucht. Dies erfolgte mithilfe des Statistikprogrammes SPSS (SPSS 20, IBM Co., Armonk, US). Es wurden Gene analysiert, die an der Biogenese des Melanins beteiligt sind (ATP7A, DTNBP1, BLOC1S5, PLDN, PMEL), die mit dem Transport und Transfer im Melanozyten assoziiert sind (RAB27A, MYO5A, MLPH), welche als Regulatoren im Melanozyten fungieren (MC1R, MITF, PAX3, SOX10) oder die über andere Mechanismen mit der Hautpigmentierung im Zusammenhang stehen (DKK1, RACK1, CNR1). Ergebnisse: Ein Kolmogorov-Smirnov-Test zeigte, dass der 25(OH)D Spiegel in der untersuchten Kohorte nicht normalverteilt ist. Aufgrund dessen wurden die weiteren statistischen Untersuchungen mit den Medianen der 25(OH)D Konzentration durchgeführt. Von den 244 getesteten erreichten 11 SNPs aus den Genen PAX3 (n=1), MLPH (n=4), BLOC1S5 (n=1), CNR1 (n=1), PMEL (n=1), ATP7A (n=3) das Signifikanzniveau (p < 0,05) und waren mit einer höheren oder niedrigeren 25(OH)D Serumkonzentration assoziiert. 3 SNPs waren nach der Korrektur für multiples Testen (nach Benjamini-Hochberg) weiterhin signifikant. Diese 3 SNPs erreichten auch das geforderte Signifikanzniveau einer Abweichung der PTH Konzentration und blieben auch nach der Korrektur für multiples Testen weiterhin signifikant. 2 SNPs aus dem Gen ATP7A zeigten zusätzlich eine Assoziation mit der Deutsche Zusammenfassung 5 1,25(OH)2D Serumkonzentration. Weiterhin wurde analysiert welcher Anteil der Varianz der 25(OH)D Konzentration von den „signifikanten“ SNPs erklärt wird. Der Determinationskoeffizient (R²) für die 11 signifikanten SNPs betrug 1,6 %. Außerdem wurden die Auswirkungen der SNPs auf die Lebenserwartung innerhalb dieser Kohorte (Patienten mit KHK) untersucht. In der Cox-Regression zeigte sich eine Assoziation des SNP rs2292881 (MLPH) mit dem Überleben. Die Kaplan-Meier Analyse ergab kein signifikantes Ergebnis. Schlussfolgerungen: Varianten von Genen, welche die Hautpigmentierung beeinflussen, haben einen Einfluss auf die 25(OH)D Konzentration des Menschen. Insgesamt zeigten sich in 6 der 15 getesteten Genen (PAX3, MLPH, BLOC1S5, CNR1, PMEL, ATP7A) Varianten, welche einen signifikanten Einfluss auf den Vitamin D Status haben.Association of variants of genes involved in skin pigmentation and vitamin D status. Background: In Caucasians, vitamin D deficiency is common and associated with higher risk for and unfavourable outcome of many diseases, including various types of cancer, infectious, cardio-vascular, and autoimmune diseases. Individual factors that predispose for a person`s vitamin D status, including skin type, have been identified, but limited data exist on genetic determinants of serum 25(OH)D concentration. Patients and Methods: In a cohort of the LURIC study, we tested the hypothesis that variants (SNPs, n=244) of several genes involved in skin pigmentation (n=15) ATP7A, DTNBP1, BLOC1S5, PLDN, PMEL (involved in melanosomal biogenesis); RAB27A, MYO5A, MLPH (encoding transfer proteins relevant for melanosomal transport within melanocytes); MC1R, MITF, PAX3, SOX10, DKK1, RACK1, CNR1 (involved in melanocyte signaling pathways) are predictive of serum 25(OH)D levels. We included 2974 patients (29.83% females, 70.17% males) with a mean 25(OH)D concentration of 17.3 ng/ml (median 15.5 ng/ml). Results: 11 SNPs located in 6 genes were associated (p < 0.05) with lower or higher serum 25(OH)D levels (medians from highest to lowest): rs6454677 (CNR1), 22.5 ng/ml, p= .046; rs2069408 (PMEL), 17.05 ng/ml, p= .015; rs2292881 (MLPH), 16.8 ng/ml, p= .041; rs7569427 (MLPH), 15.3 ng/ml, p= .026; rs9328451 (BLOC1S5), 14.6 ng/ml, p= .028; rs10932949 (PAX3), 13.9 ng/ml, p= .004; rs7565264 (MLPH), 13.1 ng/ml, p= .000992; rs17139617 (ATP7A), 12.85 ng/ml, p= .000096; rs2227291 (ATP7A), 12.8 ng/ml, p= .000047; rs10521358 (ATP7A), 12.8 ng/ml, p= .000025; rs12469812 (MLPH), 12.5 ng/ml, p= .030. 3 out of these 11 SNPs reached the aimed significance level after correction for multiple comparisons (FDR). In the linear regression model adjusted for sex, body mass index (BMI), year of birth and month of blood sample 3 SNPs showed a significant association with 25(OH)D: rs7565264 (MLPH), rs10932949 (PAX3), rs9328451 (BLOC1S5). The combined impact on the variation of 25(OH)D serum levels (coefficient of determination (R²)) for the 11 SNPs was 1.6% and for the 3 SNPs after FDR 0.3%. In Cox Regression we identified rs2292881 (MLPH) for having a significant association (advantage) with overall survival. Kaplan-Meier analysis did not show a significant impact of individual SNPs on overall survival. Conclusion: These results have a fundamental importance to understand the role of sunlight, skin pigmentation and vitamin D for the human evolution

Intratympanic application of triamcinolone in sudden hearing loss—radiologic anatomy in cone beam CT and its’ correlation to clinical outcome

Purpose!#!To evaluate temporal bone cone-beam CT in patients with idiopathic sudden sensorineural hearing loss (ISSNHL) being treated with primary and secondary intratympanic (IT) triamcinolone and to possibly correlate these results to the clinical outcome.!##!Methods!#!Retrospective analysis of patients treated with IT triamcinolone for ISSNHL at our department in 2018. Pre- and post-therapeutic audiologic examinations included four-tone average (FTA) at 0.5, 1, 2 and 3 kHz. Using a clinical questionnaire, pre-therapeutic CBCT scans were re-evaluated looking at items, which might interfere with adequate drug diffusion into the inner ear (e.g. bony overhangs or secondary membranes at the round or oval window).!##!Results!#!Thirty-one patients were included. Twenty-four (77%; group A) had experienced ineffective systemic steroid therapy before and seven (23%; group B) received primary IT injections. Four group A-patients (21%) and two group B-patients (33%) showed a post-therapeutic FTA improvement of more than 15 dB HL. Bony overhangs at the round window niche (RWN) were present in seven cases (26%), a secondary membrane at the RWN in four (15%) and soft tissue in eight (30%) cases, respectively.!##!Conclusion!#!Most patients present radiological findings in CBCT imaging, which might interfere with drug diffusion through the RW membrane. Interestingly, soft or bony tissue obstructing the RWN or the OWN was found in 50% of patients, who showed improvement of hearing. We conclude that radiologic 'tiny' findings are either clinically irrelevant or improvement in hearing is independent from intratympanic drug delivery

Monitoring of the auditory pathway maturation after early intervention during the first year of life in infants with sensorineural hearing loss

Purpose!#!The objective of this study was to investigate the auditory pathway maturation monitored by auditory brainstem responses (ABR) in infants with hearing loss during the first year of life. ABR were used to estimate hearing thresholds and the effect of early intervention strategies using hearing aids (HA).!##!Methods!#!Click-evoked ABRs were measured in 102 infants aged from 0 to 12 months to determine their individual auditory threshold. Early therapy intervention was recommended before 12 months of age and analyzed. To evaluate the effect of hearing amplification on auditory maturation, different subgroups of infants with moderate hearing loss were analyzed and the auditory pathway maturation was determined based on IPL I-V shortening.!##!Results!#!Overall, 110 ears (54.0% of 204 ears) with mild to profound HL showed threshold changes of 10 dB up to 60 dB in the follow-up ABR testing. HA were prescribed at the age of 3.8 ± 3.9 months. Cochlear implantation (CI) was performed in cases of repeated profound HL at the age of 9.9 months ± 4.5 months. A significant shortening of IPL I-V in all subgroups of infants (with and without risk factors) who received HA was shown and assumed auditory pathway maturation.!##!Conclusion!#!An early intervention using optimally fitted HA influenced auditory pathway maturation and may lead to improvements of hearing thresholds during the first year of life in infants. This study underscores the importance of not only providing HAs to infants, but also controlling for hearing threshold changes ensuring that HAs provide the optimal level of intervention or CI is indicated

First results of electrode reimplantation and its hypothetical dependence from artificial brain maturation

Background!#!After introducing the first Cochlear Implants also in children theses are grown with electrical intracochlear stimulation and subsequent auditory cortical development. Over the meantime the positioning of the electrode was changed orientated on the development of electrode design, ability to insert atraumatic and on the widening of the indications towards highfrequency deafness.!##!Methods!#!In this pilot study we analysed five prelingually deafened patients implanted as child in the late 90's and had a reimplantation 2016 or later. We compared CT and DVT (cone beam CT) scans of the temporal bone and measured the insertion angle, the cochlear coverage, the total length of the electrode in the cochlea and the distance of the first active electrode to the round window. Moreover, we compared their speech understanding before and after reimplantation.!##!Results!#!The results show a lowering in the insertion angle, the cochlear coverage, the total length of the electrode in the cochlea, in the distance of the first active electrode to the round window and in the speech understanding after reimplantation.!##!Conclusion!#!These results show a difference in the depth of insertion while the speech understanding is not significantly improving in this group-although the technology is advanced. The influence of auditory maturation with CI in these patients will be discussed

CAR-NK Cells Targeting HER1 (EGFR) Show Efficient Anti-Tumor Activity against Head and Neck Squamous Cell Carcinoma (HNSCC)

(1) Background: HNSCC is a highly heterogeneous and relapse-prone form of cancer. We aimed to expand the immunological tool kit against HNSCC by conducting a functional screen to generate chimeric antigen receptor (CAR)-NK-92 cells that target HER1/epidermal growth factor receptor (EGFR). (2) Methods: Selected CAR-NK-92 cell candidates were tested for enhanced reduction of target cells, CD107a expression and IFNγ secretion in different co-culture models. For representative HNSCC models, patient-derived primary HNSCC (pHNSCC) cell lines were generated by employing an EpCAM-sorting approach to eliminate the high percentage of non-malignant cells found. (3) Results: 2D and 3D spheroid co-culture experiments showed that anti-HER1 CAR-NK-92 cells effectively eliminated SCC cell lines and primary HNSCC (pHNSCC) cells. Co-culture of tumor models with anti-HER1 CAR-NK-92 cells led to enhanced degranulation and IFNγ secretion of NK-92 cells and apoptosis of target cells. Furthermore, remaining pHNSCC cells showed upregulated expression of putative cancer stem cell marker CD44v6. (4) Conclusions: These results highlight the promising potential of CAR-NK cell therapy in HNSCC and the likely necessity to target multiple tumor-associated antigens to reduce currently high relapse rates