Sina Jostes

Icahn School of Medicine at Mount Sinai | MSSM · Department of Oncological Sciences

High levels of H2A.Z promote melanoma cell proliferation and correlate with poor prognosis. However, the role of the two distinct H2A.Z histone chaperone complexes SRCAP and P400–TIP60 in melanoma remains unclear. Here, we show that individual subunit depletion of SRCAP , P400 , and VPS72 (YL1) results in not only the loss of H2A.Z deposition into...

High levels of H2A.Z promote melanoma cell proliferation and correlate with poor prognosis. However, the role of the two distinct H2A.Z histone chaperone complexes, SRCAP and P400-TIP60, in melanoma remains unclear. Here, we show that individual depletion of SRCAP, P400, and VPS72 (YL1) not only results in loss of H2A.Z deposition into chromatin, b...

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...

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...

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...

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...

Embryonal carcinomas (ECs) and seminomas are testicular germ cell tumours. 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 ChIP-sequencing to report and c...

Background: Type II testicular germ cell tumors (GCTs) arise from a common precursor lesion (germ cell neoplasia in situ) and are stratified into seminomas and non-seminomas, which differ considerably in morphology, gene expression, and epigenetic landscape. The N6-methyladenosine (6mA) epigenetic modification is the most abundant modification in...

Current treatment regimens for type II testicular germ cell tumours (TGCTs) achieve cure rates of ≥95%; however, 1–5% of TGCTs develop resistance to standard platinum-based chemotherapy. Patients with recurrent TGCT typically receive high-dose chemotherapy, but this treatment results in severe adverse effects and cytotoxicity. Thus, alternative tre...

Testicular germ cell tumours (GCTs) mostly affect young men at age 17-40. Although high cure rates can be achieved by orchiectomy and chemotherapy, GCTs can still be a lethal threat to young patients with metastases or therapy resistance. Thus, alternative treatment options are needed. Based on studies utilising GCT cell lines, the histone deacetyl...

Figure S2 (A) Cell morphology of JQ1 treated TGCT cell lines after 72 hrs of treatment. Scale bars: 250 μm. (B) AnnexinV/7AAD FACS analysis of apoptosis in 100 nM JQ1 treated TCam‐2 after 24 hrs, 8 days and 15 days. (C) Morphology of JQ1 treated TCam‐2 cells after 2, 4 and 16 weeks. Scale bar: 250 μm. Standard deviations, calculated by two‐tailed S...

Figure S4 (A, B) STRING‐based protein interaction prediction of genes upregulated (A) or downregulated (B) after 24 hrs of JQ1 treatment in NCCIT and TCam‐2 cells.

Figure S5 (A) qRT‐PCR validation of genes found to be deregulated in TGCT cell lines after 72 hrs of 100 nM JQ1 treatment as determined by expression microarray analysis. (B) Expression levels of deregulated genes in JQ1 treated 2102EP at higher JQ1 concentrations (750–1250 nM) as determined by qRT‐PCR analysis. Standard deviations, calculated by t...

Data S1 (A‐D) Microarray analysis of JQ1‐treated TCam‐2 (A), NCCIT (B), FS1 (C) and MPAF (D) cells after 24 and 72 hrs normalized to solvent controls. Genes commonly deregulated after 24 and 72 hrs in the respective cell lines are listed in Venn diagrams. (E) Venn diagram depicting genes commonly deregulated in TCam‐2, NCCIT, FS1 and MPAF after 24...

Figure S1 (A) BRD2, 3, 4 and T expression in human TGCT cell lines (TCam‐2, NCCIT, 2102EP), Sertoli cells (FS1) and human fibroblasts (MPAF) as determined by expression microarray analysis. (B) Quantitation of BRD2, BRD3 and BRD4 protein levels in TGCT cell lines (TCam‐2, NCCIT, NT2/D1, 2102EP) and Sertoli cells (FS1) relative to HDAC1 levels. West...

Figure S3 (A) Cell morphology of FS1 and MPAF after 72 hrs of JQ1 treatment. Scale bars: 250 μm. (B) PI‐FACS analysis of cell cycle distribution of 100 nM JQ1 treated TCam‐2 after 16 weeks. Standard deviations, calculated by two‐tailed Student's t‐test, are given above each bar.

Figure S6 (A) MYC expression in JQ1 treated TCam‐2, NCCIT, NT2/D1 and 2102EP after 24 and 72 hrs of treatment as determined by qRT‐PCR analysis. (B) MYC protein levels in JQ1 treated TGCT cell lines after 24 and 72 hrs of treatment as determined by Western blot analysis. Standard deviations, calculated by two‐tailed Student's t‐test, are given abov...

Type II testicular germ cell cancers (TGCT) are the most frequently diagnosed tumours in young men (20–40 years) and are classified as seminoma or non-seminoma. TGCTs are commonly treated by orchiectomy and chemo- or radiotherapy. However, a subset of metastatic non-seminomas (embryonal carcinomas) displays only incomplete remission or relapse and...

Protamines are arginine-rich DNA-binding proteins that replace histones in elongating spermatids. This leads to hypercondensation of chromatin and ensures physiological sperm morphology, thereby protecting DNA integrity. In mice and humans, two protamines, protamine-1 (Prm1) and protamine-2 (Prm2) are expressed in a species-specific ratio. In human...

Human germ cell development is regulated in a spatio-temporal manner by complex regulatory networks. Here, we summarize results obtained in germ cell tumors and respective cell lines and try to pinpoint similarities to normal germ cell development. This comparison allows speculating about the critical and error-prone mechanisms, which when disturbe...

Histone deacetylase inhibitor application is lethal to many cancer types. To screen for the therapeutic potential of HDIs it is necessary to analyze their ability to target and kill cancer cells in vivo. Here, we describe the xenografting of (germ cell) cancer cell lines into the flank of nude mice and the subsequent intravenous application of HDIs...

In Western countries, the incidence of testicular germ cell cancers (GCC) is steadily rising over the last decades. Mostly, men between 20 and 40 years of age are affected. In general, patients suffering from GCCs are treated by orchiectomy and radio- or chemotherapy. Due to resistance mechanisms, intolerance to the therapy or denial of chemo- / ra...

Background: Cancer/testis-antigens (CTAs) are specifically expressed in human malignancies and testis tissue, but their molecular functions are poorly understood. CTAs serve as regulators of gene expression, cell cycle and spermatogenesis, as well as targets for immune-based therapies. The CTA PRAME is expressed in various cancers, antagonises ret...

Type II germ cell cancers (GCC) are divided into seminomas, which are highly similar to primordial germ cells and embryonal carcinomas (EC), often described as malignant counterparts to embryonic stem cells. Previously, we demonstrated that the development of GCCs is a highly plastic process and strongly influenced by the microenvironment. While or...

Type II germ cell cancers (GCC) can be subdivided into seminomas and non-seminomas. Seminomas are similar to carcinoma in situ (CIS) cells, the common precursor of type II GCCs, with regard to epigenetics and expression, while embryonal carcinomas (EC) are totipotent and differentiate into teratomas, yolk-sac tumors and choriocarcinomas. GCCs can p...