18 research outputs found
Tartrate-resistant acid phosphatase/ACP5 as a driver of cancer : dissection of its oncogenic mechanisms and identification of small molecule inhibitors
No full textCancer patients diagnosed with metastasis have an increased risk of dying. To be able to predict and target tumors with an increased risk for spreading, underlying molecular events need to be better dissected and understood. TRAP is a metalloenzyme existing in two isoforms - a precursor form (TRAP 5a) and a highly enzymatically active form (TRAP 5b). TRAP expression was detected in cancer cells of several primary and metastatic tumors and expression levels were raised with increasing malignancy. TRAP expression was further correlated to clinical parameters of aggressiveness such as reduced tumor- and metastasis-free survival. Underlying molecular processes remain unclear and only a limited amount of studies is addressing the respective role of the TRAP isoforms in cancer.In this thesis, two major milestones were addressed tackling the role of TRAP (isoforms) in cancer cell metastasis. (I) Identification and characterization of previously reported and novel small molecule inhibitors of TRAP (II) Characterization of functional alterations and cellular mechanisms induced by TRAP perturbation.The TRAP inhibitor 5-phenylnicotinic acid (5-PNA/CD13) was previously identified by fragment-based screening. In Paper I, 5-PNA was further characterized for its ability to inhibit the mammalian TRAP, its selectivity for TRAP and its cytotoxicity in a cellular model. TRAP-dependent migration was inhibited by 5-PNA, shown to be selective for the TRAP 5b isoform. By small molecule screening of a library containing drug-like compounds in Paper II, several inhibitors for TRAP activity were found and selected based on a strict filtration strategy. Orthogonal validation, full-concentration responses and isoform selectivity were assessed for a selection of hit compounds. Six potential lead structures were characterized for molecular docking modes. The compound CBK289001 rendered valid as inhibiting TRAP-dependent migration in a cell system and initial structure-activity relationships were derived. The aggressiveness of cancer cells with perturbations of TRAP expression was assessed by functional studies in Paper III. TRAP had a promotive effect on cancer cell elongation, proliferation, migration and invasion. Proteomics and Phospho-proteomics outlined changes in the cellular network associated with extracellular matrix modulation and cell adhesion, and a regulation of TGFβ and CD44 signaling. A list of potential TRAP substrates was generated. The role of TRAP 5b isoform in cancer cell aggressiveness and Cathepsin K (CtsK) in its generation and processing was investigated in Paper IV. TRAP 5b was significantly increased compared to TRAP 5a in cells overexpressing TRAP. Inhibition of CtsK, an enzyme shown to be able to cleave TRAP, resulted in an intermediate processed TRAP 5b form with similar activity and promotive effect on migration. CtsK colocalized highly with TRAP 5b and cleaving changed the subcellular localization of TRAP 5b.In summary, the work presented in this thesis is contributing to the knowledge about the role of TRAP in cancer metastasis. Specifically, we were able to show a connection of TRAP 5b to metastasis-related cell functions and the involvement of TGFβ/CD44 signaling. Possible starting points for the development of potent and TRAP-specific inhibitors are provided.List of scientific papersI. Michael Krumpe, Anja Reithmeier, Teresa Senge, Toni Andreas Bäumler, Martin Frank, Per-Georg Nyholm, Barbro Ek-Rylander, Göran Andersson. The small chemical enzyme inhibitor 5-phenylnicotinic acid/CD13 inhibits cell migration and invasion of Tartrate-resistant acid phosphatase/ACP5-overexpressing MDA-MB-231 breast cancer cells. Experimental Cell Research. (2015); 339:154–162. https://doi.org/10.1016/j.yexcr.2015.09.019 II. Anja Reithmeier, Thomas Lundbäck, Martin Haraldsson, Martin Frank, Barbro Ek-Rylander, Per-Georg Nyholm, Anna-Lena Gustavsson, Göran Andersson. Identification of novel inhibitors of Tartrate-resistant acid phosphatase (TRAP/ACP5) activity by small molecule screening. [Manuscript]III. Anja Reithmeier, Elena Panizza, Michael Krumpel, Lucas Orre, Rui M. Branca, Janne Lehtiö, Barbro Ek-Rylander, Göran Andersson. Tartrate-resistant acid phosphatase (TRAP/ACP5) promotes metastasis-related cell properties via the TGFβ-pathway and CD44 in human breast cancer cells. BMC Cancer. (2017) 17:650. https://doi.org/10.1186/s12885-017-3616-7 IV. Anja Reithmeier, Tuomas Näreoja, Maria Norgård, Barbro Ek-Rylander, Göran Andersson. Tartrate-resistant acid phosphatase (TRAP/ACP5) activity promotes migration of MDA-MB-231 breast cancer cells independent of Cathepsin K processing. [Manuscript]</p
How to Become a Successful Scientist: the 2022 CSMB Arthur Wynne Gold Medal Lecture
No full textThere are many pathways to success. Mine followed a traditional one to an academic faculty position, but this pathway is not the one most life sciences PhD graduates will follow today. We have all had time during the COVID-19 Pandemic to reflect on our personal pathway -where we are and where we are going. In this reflection I outline five steps on my pathway to success:
1.Train with the best 2.Discover something 3.Mentor others 4.Go beyond 5.Promote science. I will provide examples from my personal journey that I hope will resonant with the reader as they create their pathway to success.The presentation of the authors' names and (or) special characters in the title of the pdf file of the accepted manuscript may differ slightly from what is displayed on the item page. The information in the pdf file of the accepted manuscript reflects the original submission by the author
Methoden zur Reduktion der Messlatenz von GOBO-Projektor-basierten 3D-Sensoren
No full textGenaue, optische 3D-Messverfahren werden vielfältig in der Industrie, der Medizin und der Wissenschaft eingesetzt. Bei etlichen dieser Anwendungen ist eine schnelle Reaktion auf Veränderungen in der Messszene erforderlich. Dies ist z.B. der Fall, wenn eine Maschine aus Sicherheitsgründen abgeschaltet oder angehalten werden muss oder eine direkte Rückmeldung an einen Menschen gegeben werden soll. Steht das 3D-Ergebnis der Messung nach hinreichend kurzer Zeit ab Beginn der Messung zur Verfügung, kann die Reaktion basierend auf diesem Ergebnis angestoßen werden. Das Prinzip des GOBO-Projektor-basierten, aktiven Stereo-Sensors hat sich als genaues, optisches 3D-Messverfahren etabliert. Bei diesem Verfahren wird ein sich zeitlich änderndes, aperiodisches Streifenmuster auf das Messobjekt projiziert, während zwei kalibrierte Kameras jeweils eine Bildsequenz synchron aufnehmen. Innerhalb dieser Bildsequenzen werden anschließend die Abbilder von Objektpunkten, welche in beiden Kameras sichtbar sind, einander zugeordnet. Für jedes solche Paar wird dann die 3D-Koordinate des zugehörenden Objektpunktes trianguliert. Das Verfahren erlaubt auch 3D-Aufnahmen mit speziellen Anforderungen, die von anderen, genauen 3D-Sensorprinzipien nur schwer erreicht werden. Dazu gehört die Messung mit speziellen Lichtwellenlängen, wie z.B. dem Nah-Infrarotbereich, womit blendfreie Vermessung ermöglicht wird, oder die Erfassung sehr schneller Prozesse, wie die Messung von Airbag-Entfaltungen. Bisher war es jedoch nicht möglich, die 3D-Messergebnisse in so kurzer Zeit (z.B. 100 ms), d.h. mit so kurzer Messlatenz, zur Verfügung zu stellen, dass eine unmittelbare Reaktion auf eine Veränderung der Messszene erfolgen kann. Diese Verkürzung der Messlatenz ist das Ziel dieser Arbeit. Es werden Methoden beschrieben und untersucht, mit denen die Messlatenz von GOBO-Projektor-basierten, aktiven Stereo-Sensoren auf unter 100 ms verkürzt werden kann. Die Verbesserungen konzentrieren sich auf zwei Bereiche: die schnelle Rekonstruktion des 3D-Modells aus den aufgenommenen Bildsequenzen und die Reduktion der Aufnahmezeit durch Verkürzung der Bildsequenz-Länge. Letztere wird mittels einer Optimierung der Musterprojektion ermöglicht, welche bei kurzen Bildsequenz-Längen eine erhebliche Reduktion unerwünschter Messartefakte bewirkt. Abschließend werden mehrere Anwendungen gezeigt, die von diesen Verbesserungen profitieren.Accurate, optical 3D measurement techniques are widely used in industry, medicine and science. In some of the applications, a fast response to changes in the measurement scene is required. This is the case, for example, when a machine has to be stopped to avoid collision, or direct feedback to a human being must be provided. If the 3D result of the measurement is available after a sufficiently short time from the start of the measurement, the reaction can be triggered based on this result. Active stereo 3D sensors based on GOBO projection have been established as an accurate optical 3D measurement method. The sensor projects a time-varying, aperiodic fringe pattern onto the measured object, while two calibrated cameras synchronously record an image sequence, each. After the recording, a 3D reconstruction algorithm searches for pixels pairs, which correspond to the same object point within these two image sequences. For each such pair, the algorithm triangulates the 3D coordinate of the associated object point. With this method, special requirements can be accounted for which are difficult to achieve by other accurate 3D measurement principles. These include measurements with special light wavelengths, such as the near-infrared range, thus enabling glarefree, i.e. irritation-free measurement, or the measurement of very fast processes, such as the measurement of airbag inflation. Until now, however, it has not been possible to provide 3D measurement results in such a short time (e.g. 100ms), i.e. with such a short measurement latency, that an immediate reaction to a change in the measurement scene can be made. Therefore, the goal of this dissertation is the reduction of the measurement latency. In this dissertation, the author describes and investigates methods to shorten the measurement latency of GOBO projector-based active stereo sensors to less than 100ms. The improvements focus on two areas: fast reconstruction of the 3D model from the acquired image sequences, and reduction of the acquisition time by shortening the image sequence length. The latter is achieved by means of an optimization of the pattern projection, which results in a significant reduction of unwanted measurement artifacts for short image sequence lengths. Finally, the author shows several applications that benefit from these improvements
A Planar low-cost full-polymer Optical Humidity Sensor
No full textAbstractWe present an all-polymer optical humidity sensor, based on a 1mm plastic optical fiber (POF) with a U-bend, cladded with poly(N,N-dimethylacrylamide) (PDMAA) in the sensing region. The cladding changes its scattering properties on absorption of environmental humidity, thus modulating the transmitted optical power through the sensor. We explain the working principle of the sensor and show experimental results regarding scattering behavior of the cladding material and sensitivity to sudden humidity changes. We also propose a planar layout suitable for application to a hot embossing or lamination process for large-scale fabrication
Validating Signal Transducer and Activator of Transcription (STAT) ProteinInhibitor Interactions Using Biochemical and Cellular Thermal Shift Assays
No full textAdditional file 4 of Cathepsin K regulates localization and secretion of Tartrate-Resistant Acid Phosphatase (TRAP) in TRAP-overexpressing MDA-MB-231 breast cancer cells
No full textAdditional file 4: Supplementary video 3 mock ctrl totalTRAP. Z-sections through ICC-stained, proCtsK (Alexa 488, green) and total TRAP (Alexa 647, magenta), mock control cells
Additional file 2 of Cathepsin K regulates localization and secretion of Tartrate-Resistant Acid Phosphatase (TRAP) in TRAP-overexpressing MDA-MB-231 breast cancer cells
No full textAdditional file 2: Supplementary video 1 mock ctrl TRAP 5a. Z-sections through ICC-stained proCtsK (Alexa 488, green) and uncleaved TRAP 5a (Alexa 647, magenta) mock control cells displayed in Fig. 3 as a maximum intensity projection
Additional file 5 of Cathepsin K regulates localization and secretion of Tartrate-Resistant Acid Phosphatase (TRAP) in TRAP-overexpressing MDA-MB-231 breast cancer cells
No full textAdditional file 5: Supplementary video 4 TRAPhi totalTRAP. Z-sections through ICC-stained, proCtsK (Alexa 488, green) and total TRAP (Alexa 647, magenta), TRAP3high cells
Additional file 3 of Cathepsin K regulates localization and secretion of Tartrate-Resistant Acid Phosphatase (TRAP) in TRAP-overexpressing MDA-MB-231 breast cancer cells
No full textAdditional file 3: Supplementary video 2 TRAPhi TRAP 5a. Z-sections through ICC-stained proCtsK (Alexa 488, green) and uncleaved TRAP 5a (Alexa 647, magenta) TRAP3high cells displayed in Fig. 3 as a maximum intensity projection
Additional file 1 of Cathepsin K regulates localization and secretion of Tartrate-Resistant Acid Phosphatase (TRAP) in TRAP-overexpressing MDA-MB-231 breast cancer cells
No full textAdditional file 1: Supplementary material. Including co-staining of TRAP isoforms and proCtsK in MDA-MB-231 cells treated with MK-0822. Densitometric quantification of Western blot TRAP bands of the different FPLC fractioned cell lysates. Controls for nonspecific binding of secondary antibodies
