SP Cancer Metastasis Summary Hypothesis: We hypothesize that miRNAs regulate breast cancer cell invasiveness and metastasis by synergistically targeting.

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SP Cancer Metastasis Summary Hypothesis: We hypothesize that miRNAs regulate breast cancer cell invasiveness and metastasis by synergistically targeting several metastasis-related phenotypes and pathways. Such miRNAs might serve on the one hand as diagnostic or prognostic markers, but might on the other hand also provide valuable information on novel strategies to fight metastasis formation by interfering with the corresponding signalling pathways.

SP Cancer Metastasis Previous work Impact of certain miRNAs on breast cancer development and progression Functional analysis of miRNA effects on signaling pathways and cellular phenotypes –e.g. Keklikoglou et al: miR-520c as metastasis suppressor in ER - –e.g. Uhlmann et al: miR-200c as tumor suppressor Relevant tools –High-content screening microscope –Real-time cell analyzer (RTCA) –96-well pipetting robot for automized screening

SP Cancer Metastasis Relevant expertise Identification and functional characterization of miRNA targets Integration of cancer patient data to prove clinical significance of miRNA/target interactions Set-up and conduction of large-scale screenings (so far siRNA/miRNA screens) Cellular assays to analyze cell migration and invasion in vitro –Automated wound-healing scratch assay –RTCA-based and conventional transwell migration and invasion assays –3D cell culture model for invasive growth (MCF10A)

WP/Aim 1: We will identify miRNAs and their target genes with clinical relevance in subtypes of breast cancer. WP/Aim 2: We will identify clinically relevant and prognostic SNPs or mutations within the seed regions of target genes. WP/Aim 3: We will identify and functionally characterize miRNAs impacting on cancer cell invasion in vitro and in vivo. WP/Aim 4: We will verify clinical significance of the identified miRNA target signatures using patient samples as well as publicly available data sets on miRNA and mRNA and protein expression. WP/Aim 5: We will identify compounds modulating metastasis-related phenotypes in vitro and metastasis formation in vivo in similar ways as the potent miRNAs identified in this project. SP Cancer Metastasis Workpackages

Aim 3,5 Aim 4,5 Aim 3,5 Aim 2 SP Cancer Metastasis Internal Networking Aim 1,4 Aim 1,3