A5051147274- PARP inhibition in cancer therapy
- RNA Interference and Gene Delivery
- Calcium signaling and nucleotide metabolism
- Melanoma and MAPK Pathways
- RNA Research and Splicing
- CRISPR and Genetic Engineering
- CAR-T cell therapy research
- Nuclear Structure and Function
- Corneal Surgery and Treatments
- Skin and Cellular Biology Research
- Genomics and Chromatin Dynamics
- Cell death mechanisms and regulation
- Virus-based gene therapy research
- Toxin Mechanisms and Immunotoxins
- Protein Degradation and Inhibitors
- Microbial Metabolic Engineering and Bioproduction
- DNA Repair Mechanisms
- Autophagy in Disease and Therapy
- Animal Genetics and Reproduction
- Advancements in Transdermal Drug Delivery
- Retinal Development and Disorders
- Cardiac electrophysiology and arrhythmias
- Bioinformatics and Genomic Networks
- RNA and protein synthesis mechanisms
- Ocular Disorders and Treatments
University of Zurich
2016-2024
University of Dundee
2012-2020
University Hospital of Zurich
2020
ETH Zurich
2010
École Polytechnique Fédérale de Lausanne
2008
Oregon Health & Science University
2006
Institute for Systems Biology
2002-2006
University of Washington
2005-2006
Vanderbilt University
2006
University of Alberta
2002-2004
Abstract Objective MicroRNAs (miRNA) have recently emerged as a new class of modulators gene expression. In this study we investigated the expression, regulation, and function miR‐155 miR‐146a in rheumatoid arthritis (RA) synovial fibroblasts (RASFs) RA tissue. Methods Locked nucleic acid microarray was used to screen for differentially expressed miRNA RASFs treated with tumor necrosis factor α (TNFα). TaqMan‐based real‐time polymerase chain reaction applied measure levels miR‐146a. Enforced...
Different experimental technologies measure different aspects of a system and to differing depth breadth. High-throughput assays have inherently high false-positive false-negative rates. Moreover, each technology includes systematic biases nature. These differences make network reconstruction from multiple data sets difficult error-prone. Additionally, because the rapid rate progress in biotechnology, there is usually no curated exemplar set which one might estimate integration parameters....
CRISPR/Cas9-based therapeutics hold the possibility for permanent treatment of genetic disease. The potency and specificity this system has been used to target dominantly inherited conditions caused by heterozygous missense mutations through inclusion mutated base in short-guide RNA (sgRNA) sequence. This research evaluates a novel approach targeting single-nucleotide polymorphisms (SNPs) using CRISPR/Cas9. We determined that mutation within KRT12, which causes Meesmann's epithelial corneal...
The integration of data from multiple global assays is essential to understanding dynamic spatiotemporal interactions within cells. In a companion paper, we reported methodology, designated Pointillist, that can handle types technologies with different noise characteristics. Here demonstrate its application the 18 sets relating galactose utilization in yeast. These include changes mRNA and protein abundance, genome-wide protein–DNA interaction data, database information, computational...
The lymphatic vascular system maintains tissue fluid homeostasis, helps mediate afferent immune responses, and promotes cancer metastasis. To address the role microRNAs (miRNAs) play in development function of system, we defined vitro miRNA expression profiles primary human endothelial cells (LECs) blood (BVECs) identified four BVEC signature two LEC miRNAs. Their lineage-specific patterns were confirmed vivo by quantitative real-time PCR situ hybridization. Functional characterization...
Keratin 9 (K9) is a type I intermediate filament protein whose expression confined to the suprabasal layers of palmoplantar epidermis. Although mutations in K9 gene are known cause epidermolytic keratoderma, rare dominant-negative skin disorder, its functional significance poorly understood. To gain insight into physical requirement and importance K9, we generated K9-deficient (Krt9(-/-)) mice. Here, report that adult Krt9(-/-)mice develop calluses marked by hyperpigmentation exclusively...
Abstract Diverse cellular processes depend on the lysosomal protease system but how cells regulate proteolytic capacity is only partly understood. We show here that can respond to protease/substrate imbalance in this compartment by de novo expression of multiple hydrolases. This response, exemplified either loss asparagine endopeptidase (AEP) or other cysteine proteases, increased endocytic substrate load, not dependent transcription factor EB (TFEB) rather triggered STAT3 activation...
Abstract Protein ADP-ribosylation is a reversible post-translational modification that regulates important cellular functions. The identification of modified proteins has proven challenging and mainly been achieved via enrichment methodologies. Random mutagenesis was used here to develop an engineered Af1521 ADP-ribose binding macro domain protein with 1000-fold increased affinity towards ADP-ribose. crystal structure reveals two point mutations K35E Y145R form salt bridge within the domain....
In addition to its role as an electron transporter, mitochondrial nicotinamide adenine dinucleotide (NAD+) is important co-factor for enzymatic reactions, including ADP-ribosylation. Although mitochondria harbor the most intra-cellular NAD+, ADP-ribosylation remains poorly understood. Here we provide evidence ADP-ribosylation, which was identified using various methodologies immunofluorescence, western blot, and mass spectrometry. We show that reversibly increases in response respiratory...
Though the effect of recently identified mitochondrial NAD+ transporter SLC25A51 on glucose metabolism has been described, its contribution to other NAD+-dependent processes throughout cell such as ADP-ribosylation remains elusive. Here, we report that absence leads increased concentration not only in cytoplasm and but also nucleus. The increase is associated with upregulation salvage pathway, implying an accumulation constitutively synthesized This results PARP1-mediated nuclear...
PARP7 was reported to promote tumor growth in a cell-autonomous manner and by repressing the antitumor immune response. Nevertheless, molecular mechanism of how PARP7-mediated ADP-ribosylation exerts these effects cancer cells remains elusive. Here, we identified as nuclear cysteine-specific mono-ADP-ribosyltransferase that modifies targets critical for regulating transcription, including AP-1 transcription factor FRA1. Loss FRA1 via inhibition RBN-2397 or mutation site C97 increased...
Abstract Metastatic melanoma is either intrinsically resistant or rapidly acquires resistance to targeted therapy treatments, such as MAPK inhibitors (MAPKi). A leading cause of a dynamic transition cells from proliferative highly invasive state, phenomenon called phenotype switching. Mechanisms regulating switching represent potential targets for improving treatment patients with melanoma. Using drug screen targeting chromatin regulators in patient-derived three-dimensional MAPKi-resistant...
To identify an allele-specific short interfering RNA (siRNA), against the common KRT12 mutation Arg135Thr in Meesmann epithelial corneal dystrophy (MECD) as a personalized approach to treatment.siRNAs K12 were evaluated using dual luciferase reporter gene assay and most potent specific siRNAs further screened by Western blot. Off-target effects on related keratins assessed immunological stimulation of TLR3 was RT-PCR. A modified 5' rapid amplification cDNA ends method used confirm...
Therapeutics based on short interfering RNAs (siRNAs), which act by inhibiting the expression of target transcripts, represent a novel class potent and highly specific next-generation treatments for human skin diseases. Unfortunately, intrinsic barrier properties combined with large size negative charge siRNAs make epidermal delivery these macromolecules quite challenging. To help evaluate in vivo activity therapeutics refine strategies we generated an innovative reporter mouse model that...
In yeast there are at least 14 members of the beta-karyopherin protein family that govern movement a diverse set cargoes between nucleus and cytoplasm. Knowledge carried by each karyopherin insight into mechanisms transport fundamental to understanding constitutive regulated elucidating how they impact normal cellular functions. Here, we have focused on identification nuclear import for essential beta-karyopherin, Kap121p. Using an overlay blot assay coimmunopurification studies, identified...
The aim of this study is to further assess our previously reported keratin 12 (K12)-Leu132Pro specific siRNA in silencing the mutant allele Meesmann's Epithelial Corneal Dystrophy (MECD) experimental systems more akin vivo situation through simultaneous expression both wild-type and alleles.Using KRT12 exogenous constructs transfected into cells, knockdown was quantified using pyrosequencing infrared Western blot analysis, while mechanism assessed by a modified rapid amplification cDNA ends...
Tumor necrosis factor (TNF) is a key component of the innate immune response. Upon binding to its receptor, TNFR1, it promotes production other cytokines via membrane-bound complex 1 or induces cell death cytosolic 2. To understand how TNF-induced regulated, we performed mass spectrometry 2 and identified tankyrase-1 as native that, upon stimulus, mediates poly-ADP-ribosylation (PARylation). PARylation recruitment E3 ligase RNF146, resulting in proteasomal degradation 2, thereby limiting...
To further our understanding of how the nucleocytoplasmic transport machinery interfaces with its cargoes and this affects cellular physiology, we investigated molecular mechanisms phenotypes associated mutations in karyopherin Kap121p. Two previously unreported kap121 cells were observed: defects mating transition from normal yeast form to pseudohyphal, invasive form. In parallel, searched for Kap121p by using as a probe overlay assays nuclear proteins. One major interacting proteins...
Meesmann epithelial corneal dystrophy (MECD) is a rare autosomal dominant disorder caused by dominant-negative mutations within the KRT3 or KRT12 genes, which encode cytoskeletal protein keratins K3 and K12, respectively. To investigate pathomechanism of this disease, we generated phenotypically characterized novel knock-in humanized mouse model carrying severe, MECD-associated, K12-Leu132Pro mutation. Although no overt changes in opacity were detected slit-lamp examination, corneas...
A yeast tRNA three-hybrid interaction approach and an in vivo nuclear export assay based on amber suppression was used to identify proteins that participate the process Saccharomyces cerevisiae. One of identified by this strategy is Utp8p, essential 80-kDa nucleolar protein has been implicated 18 S ribosomal RNA biogenesis. Our characterization indicated major function Utp8p export. Like S. cerevisiae Los1p mammalian exportin-t, which are known facilitate export, overexpression restored...
Abstract Pluripotency is established in E4.5 preimplantation epiblast. Embryonic stem cells (ESCs) represent the immortalization of pluripotency, however, their gene expression signature only partially resembles that developmental ground-state. Induced PRAMEL7 expression, a protein highly expressed ICM but lowly ESCs, reprograms developmentally advanced ESC+serum into ground-state pluripotency by inducing close to However, how remains elusive. Here we show associates with Cullin2 (CUL2) and...