Daniel Nettersheim

Heinrich-Heine-Universität Düsseldorf | HHU · Department of Urology, Urological Research Laboratory, Translational Uro-Oncology, University Hospital Düsseldorf

Urologic malignancies represent major challenges for clinicians with annually rising incidences. Additionally, cisplatin treatment-induced long-term toxicities and the development of therapy resistance emphasize the need for novel therapeutics. In this study, we analyzed the effects of novel histone deacetylase (HDAC) and bromodomain and extratermi...

Alternative therapeutic options targeting urologic malignancies, such as germ cell tumours, as well as urothelial, renal and prostate carcinomas, are still urgently needed. The membrane protein CD24 represents a promising immunotherapeutical approach. The present study aimed to decipher the molecular function of CD24 in vitro and evaluate the cytot...

Background: Germ cell tumors (GCT) might undergo transformation into a somatic-type malignancy (STM), resulting in a cell fate switch to tumors usually found in somatic tissues, such as rhabdomyosarcomas or adenocarcinomas. STM is associated with a poor prognosis, but the molecular and epigenetic mechanisms triggering STM are still enigmatic, the...

In germ cell tumors (GCT), a growing teratoma during chemotherapy with decreasing tumor markers was defined as ‘growing teratoma syndrome` (GTS) by Logothetis et al., in 1982. So far, its pathogenesis and specific treatment options remain elusive. We aimed at updating the GTS definition based on molecular and epigenetic features as well as identify...

Germ cell tumors (GCT) are the most common solid tumors in young men of age 15 - 40. In previous studies, we profiled the interaction of GCT cells with cells of the tumor microenvironment (TM), which showed that especially the 3D interaction of fibroblasts (FB) or macrophages with GCT cells influenced the growth behavior and cisplatin response as w...

Testicular germ cell tumours (GCT) are divided into three different subtypes (types I–III) regarding to their developmental origin, histological differences and molecular features. Type I GCT develop from disturbed primordial germ cells and most commonly occur in children and young adolescents, which is why they are referred to as prepubertal GCT....

Background: Being implicated during tumor migration, invasion, clonogenicity, and proliferation, the nicotinamide adenine dinucleotide (NAD) / -phosphate (NADP)-dependent dehydrogenase / reductase member 2 (DHRS2) has been considered to be induced upon inhibition of histone deacetylases (HDACi). In this brief report, we evaluated the current knowle...

Meriolin derivatives represent a new class of kinase inhibitors with a pronounced cytotoxic potential. Here, we investigated a newly synthesized meriolin derivative (termed meriolin 16) that displayed a strong apoptotic potential in Jurkat leukemia and Ramos lymphoma cells. Meriolin 16 induced apoptosis in rapid kinetics (within 2–3 h) and more pot...

Approximately 30% of seminoma (SEM) patients present with moderately elevated human chorionic gonadotropin (hCG) levels at first diagnosis. In case of high hCG serum levels, the presence of a non-SEM component, i.e. choriocarcinoma (CC), may be assumed. To characterize cases described as pure seminoma with high serum hCG levels, tissue samples and...

Being stimulated by the chemokine CXCL12, the CXCR4 / CXCR7 cascade is involved in tumor proliferation, migration, and metastasis. The interaction between CXCL12, secreted by cells from the microenvironment, and its receptors is complex and has been ascribed to promote chemotherapy resistance. However, the role of this signaling axis and its target...

Meriolin derivatives represent a new class of kinase inhibitors with a pronounced cytotoxic potential. Here, we investigated a newly synthesized meriolin derivative (termed meriolin 16) that displayed a strong apoptotic potential in Jurkat leukemia and Ramos lymphoma cells. Meriolin 16 induced apoptosis in rapid kinetics (within 2 - 3 h) and more p...

Aims Testicular germ cell tumours are the most common solid malignancies in young men of age 14–44 years. It is generally accepted that both seminomas and non‐seminomas arise from a common precursor, the germ cell neoplasia in‐situ , which itself is the result of a defective (primordial) germ cell development. The stem cell‐like population of the n...

Background Type II testicular germ cell tumours (TGCT) are the most prevalent tumours in young men. Patients suffering from cisplatin-resistant TGCTs are facing very poor prognosis demanding novel therapeutic options. Neddylation is a known posttranslational modification mediating many important biological processes, including tumorigenesis. Overac...

Background: Being the standard-of-care for four decades, cisplatin-based chemotherapy is highly efficient in treating germ cell tumors (GCT). However, often refractory patients present with a remaining (resistant) yolk-sac tumor (YST(-R)) component, resulting in poor prognosis due to lack of novel treatment options besides chemotherapy and surgery...

Background: Type II germ cell tumors (GCT) are the most common solid cancers in males of age 15 to 35 years. Treatment of these tumors includes cisplatin-based therapy achieving high cure rates, but also leading to late toxici-ties. As mainly young men are suffering from GCTs, late toxicities play a major role regarding life expectancy, and the de...

Background Germ cell tumors (GCTs) are the most common type of cancer in Germany in young men between 15 and 44 years of age. The routinely performed diagnostic procedures are essential for the patient’s treatment, but can be difficult due to heterogenous histologies. Additionally, the molecular mechanisms of the development of the special forms gr...

The tumor microenvironment (TM), consisting of the extracellular matrix (ECM), fibroblasts, endothelial cells and immune cells, might affect tumor invasiveness and the outcome of standard chemotherapy. This study investigated the cross‐talk between germ cell tumors (GCT) and surrounding TM cells (macrophages, T‐lymphocytes, endothelial cells, fibro...

Germ cell tumors of the testis are rare cancers with-for unknown reasons-rising incidence in industrialized countries affecting young men in their reproductive ages peaking at 20-30 and 30-40 years depending on histology. These special circumstances demand different diagnostic and monitoring tools, different counseling and different multidisciplina...

Purpose Clinical stage I (CSI) testicular germ cell tumors (TGCT) represents disease confined to the testis without metastasis and CSIS is defined as persistently elevated tumor markers (TM) after orchiectomy, indicating subclinical metastatic disease. This study aims at assessing clinical characteristics and oncological outcome in CSIS. Methods D...

Purpose Advanced testicular germ cell tumours (GCT) generally have a good prognosis owing to their unique sensitivity towards cisplatin-based chemotherapies. However, cisplatin-resistant GCT have a poor outcome. Further studies are mandatory to better understand resistance mechanisms and develop therapeutic strategies for refractory GCTs. Methods...

Background Germ cell tumors (GCTs) are developmental cancers, tightly linked to embryogenesis and germ cell development. The recent and expanding field of RNA modifications is being increasingly implicated in such molecular events, as well as in tumor progression and resistance to therapy, but still rarely explored in GCTs. In this work, and as a f...

Aims Primary mediastinal germ cell tumours (PMGCTs) are rare mediastinal neoplasms and their diagnosis can be challenging due to small biopsy samples. The aim of this study was to elaborate a diagnostic algorithm using immunohistochemical stainings with focus on novel markers and molecular analysis of isochromosome 12p [i(12p)]. Methods and result...

Testicular germ cell tumors (GCTs) are stratified into seminomas and non‐seminomas. Seminomas share many histological and molecular features with primordial germ cells, whereas the non‐seminoma stem cell population – embryonal carcinoma (EC) – is pluripotent and thus able to differentiate into cells of all three germ layers (teratomas). Furthermore...

Germ cell neoplasia in situ (GCNIS) is the noninvasive precursor of testicular germ cell tumors type II, the most common cancer in young men, which originates from embryonic germ cells blocked in their maturation. GCNIS is associated with impaired Sertoli cells (SCs) that express fetal keratin 18 (KRT18) and the pluripotency factor SRY-Box transcri...

Cisplatin-based chemotherapy has been used for more than four decades as a standard therapeutic option in several tumor entities. However, being a multifaceted and heterogeneous phenomenon, inherent or acquired resistance to cisplatin remains a major obstacle during the treatment of several solid malignancies and inevitably results in disease progr...

Yolk-sac tumours (YSTs), a germ cell tumour subtype, occur in newborns and infants as well as in young adults of age 14-44 years. In clinics, adult patients with YSTs face a poor prognosis, as these tumours are often therapy-resistant and count for many germ cell tumour related deaths. So far, the molecular and (epi)genetic mechanisms that control...

Cisplatin resistance still remains a major obstacle in the standard chemotherapeutic approach in late-stage and metastatic testicular germ cell cancer (GCC) patients. This multifactorial and complex phenomenon arises (concomitantly) on several levels due to impaired transport, decreased adduct formation, increased DNA-repair, decreased apoptosis, o...

Type II testicular germ cell tumors (GCTs) can be classified as seminoma or embryonal carcinoma. Both subtypes present distinct cellular morphologies and characteristics. Seminomas closely resemble primordial germ cells (PGCs) with respect to their transcriptome and epigenetic signature (DNA hypomethylation). They express the pluripotency markers L...

The hanging drop cell culture technique allows to study three-dimensional growth and differentiation of cell aggregates, that is, embryonic stem cells. Compared to standard two-dimensional monolayer cell cultivation, hanging drops allow for a better visualization and understanding of the developmental processes in vitro. Hanging drop cultivation ca...

Background Despite excellent cure rates due to platinum-based combination chemotherapy in advanced stages, 10–15% of germ cell tumour (GCT) patients display a cisplatin-refractory disease and have a dismal prognosis.Objectives An overview on the molecular background, current treatment approaches, and potential future options for cisplatin refractor...

Background The diagnosis of growing teratoma syndrome (GTS) is complex and due to the small number of cases it is difficult to evaluate the treatment options. A better understanding of the development of GTS and sensitization to its paradoxical clinical presentation is crucial for a successful treatment strategy.Objectives The aim of this review is...

Filamins are large dimeric F-actin cross-linking proteins, crucial for the mechanosensitive properties of a number of cell types. Due to their interaction with a variety of different proteins, they exert important regulatory functions. However, in the human testis the role of filamins has been insufficiently explored. Immunohistochemical staining o...

Simple Summary There is a need for novel treatment options for patients with testicular germ cell tumors, especially for those that are resistant to standard chemotherapy, who show poor prognosis. In this work, we test two compounds that inhibit epigenetic enzymes called histone deacetylases—belinostat and panobinostat. We show that these enzymes a...

Aims Malignant germ cell tumours (GCTs) of the testis are rare neoplasms, but the most common solid malignancies in young men. World Health Organization guidelines divide GCTs into five types, for which numerous immunohistochemical markers allow exact histological subtyping in the majority of cases. In contrast, a germ cell origin is often hard to...

Background/aim: Myoferlin (MYOF) has emerged as an oncogenic protein in various human cancer types. This study was conducted to investigate comprehensively the expression and functional properties of MYOF in clear-cell renal-cell carcinoma (ccRCC) with respect to its value as diagnostic biomarker and therapeutic target. Materials and methods: mR...

The development of germ cell tumors (GCTs) is a unique pathogenesis occurring at an early developmental stage during specification, migration or colonization of primordial germ cells (PGCs) in the genital ridge. Since driver mutations could not be identified so far, the involvement of the epigenetic machinery during the pathogenesis seems to play a...

Outcome in high-risk patients with refractory or relapsed germ cell tumours (GCT) remains poor. Novel strategies enhancing therapeutic efficacy whilst limiting therapeutic burden are warranted, yet immunotherapy approaches geared towards activating endogenous antitumor responses have not been successful thus far. Redirection of cytotoxic effector c...

Background The vesicle fusion protein Dysferlin (DYSF) is mainly known as a membrane repair protein in muscle cells. Mutations of DYSF lead to muscular dystrophies and cardiomyopathies. In contrast to other members of the Ferlin protein family, few is known about its role in cancer. Our study was designed to investigate the expression and functiona...

Germ cell tumours (GCTs) are the most common solid malignancies in young men. Although high cure rates can be achieved, metastases, resistance to cisplatin-based therapy and late toxicities still represent a lethal threat, arguing for the need of new therapeutic options. In this study, we analysed the potential of cyclin-dependent kinase 4/6 (CDK4/...

Germ cell tumors (GCTs) are the most common solid malignancies found in young men. Although they generally have high cure rates, metastases, resistance to cisplatin-based therapy, and late toxicities still represent a lethal threat, arguing for the need of new therapeutic options. In a previous study, we identified downregulation of the chromatin-r...

Embryonal carcinomas (ECs) and seminomas are testicular germ cell tumors. ECs display expression of SOX2, while seminomas display expression of SOX17. In somatic differentiation, SOX17 drives endodermal cell fate. However, seminomas lack expression of endoderm markers, but show features of pluripotency. Here, we use chromatin immunoprecipitation se...

Abstract of a presentation given by Daniel Nettersheim at the International Extracranial Germ Cell Tumor Conference 2019 in Cambridge, UK. Published at:

Figure S3: A SOX2- and SOX2/FOXA2-deficiency does not influence the seminoma-like cell fate of TCam-2 cells in vitro. (A,B) qRT-PCR (A) and western blot (B) analysis of indicated marker genes in parental TCam-2, TCam-2-deltaSOX2 and TCam-2-deltaSOX2/FOXA2 cells. 2102EP and PC3 cells served as additional controls. Densitometrical analysis of western...

Figure S4: HE and IHC staining of SOX2, FOXA2, the pluripotency and seminoma markers OCT4, SOX17, TFAP2C and PRDM1 (green), the differentiation markers AFP and EOMES (red) and the proliferation marker Ki67 (blue) in TCam-2-SOX2/FOXA2 tumor tissues twelve weeks after xenografting.

Figure S5: Expression of FOXA2 in GCT cell lines and tissues as well as supplemental western blot data. (A,B) Microarray expression analysis of FOXA2 expression in GCT cell lines (A) and tissues (B). Black lines indicate the minimum level of expression intensity that can be considered as ‘expressed’. Differences in these thresholds is because of us...

Testicular germ cell tumors (GCTs) are very common in young men and can be stratified into seminomas and non-seminomas. While seminomas share a similar gene expression and epigenetic profile with primordial germ cells, the stem cell population of the non-seminomas, the embryonal carcinoma (EC), resembles malignant embryonic stem cells. Thus, ECs ar...

Figure S2: Measurement of proliferation rates of parental TCam-2, TCam-2-deltaSOX2 and TCam-2-deltaSOX2/FOXA2 cells over eleven days.

Figure S1: Validation of a successful FOXA2 gene editing. (A) Schematic of the CRISPR/Cas9 strategy to knock out FOXA2 in TCam-2 cells. Location of guideRNA1 - 3 and primer pairs for genotyping PCR are depicted. (B) Genotyping PCR using all three primer pairs depicted in A). Three clones, already deficient for SOX2 [9], show also deficiency for FOX...

Am 24. Januar 2019 fand auf Schloss Mickeln, dem Tagungshaus der Heinrich-Heine-Universität Düsseldorf, der erste „Düsseldorfer Testis Cancer Day“ (DusTCD) statt. Mit dem Ziel Grundlagenforscher und klinisch-/wissenschaftlich-tätige Ärzte mit Schwerpunkt in der Erforschung und Behandlung von Keimzelltumoren (KZT) zusammenzubringen, trafen sich ca....