Impact of cathepsin B on the interstitial fluid proteome of murine breast cancers

Gomez-Auli, Alejandro; Hillebrand, Larissa E.; Biniossek, Martin L.; Peters, Christoph; Reinheckel, Thomas; Schilling, Oliver

doi:10.1016/j.biochi.2015.10.009

Cited by 23 publications

(19 citation statements)

References 54 publications

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“…5 j), consistent with the preferential binding of the DARPin to this CatB form due to a steric clash with the propeptide that results in a diminished rate of association and affinity for the zymogen. This is in agreement with the recent results of a proteomic analysis of PyMT tumour interstitial fluids, which showed that at least part of the secreted CatB exists in the zymogen proCatB form 63 . Our results therefore support previous findings, where a significant proportion of the secreted zymogen has been found to be activated in the acidic extracellular milieu in tumours 64 , 65 .…”

Section: Discussionsupporting

confidence: 93%

Non-invasive in vivo imaging of tumour-associated cathepsin B by a highly selective inhibitory DARPin

et al. 2017

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Cysteine cathepsins often contribute to cancer progression due to their overexpression in the tumour microenvironment and therefore present attractive targets for non-invasive diagnostic imaging. However, the development of highly selective and versatile small molecule probes for cathepsins has been challenging. Here, we targeted tumour-associated cathepsin B using designed ankyrin repeat proteins (DARPins). The selective DARPin 8h6 inhibited cathepsin B with picomolar affinity (Ki = 35 pM) by binding to a site with low structural conservation in cathepsins, as revealed by the X-ray structure of the complex. DARPin 8h6 blocked cathepsin B activity in tumours ex vivo and was successfully applied in in vivo optical imaging in two mouse breast cancer models, in which cathepsin B was bound to the cell membrane or secreted to the extracellular milieu by tumour and stromal cells. Our approach validates cathepsin B as a promising diagnostic and theranostic target in cancer and other inflammation-associated diseases.

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Section: Discussionsupporting

confidence: 93%

Non-invasive in vivo imaging of tumour-associated cathepsin B by a highly selective inhibitory DARPin

et al. 2017

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“…To determine differentially regulated proteins, we performed a linear model for microarrays (limma) fitting, which has been previously used for quantitative proteomic data 13 , 31 . The corresponding volcano plots are depicted in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The FC values were normalized to center at zero. Statistical significance was tested using the linear model for microarray data (limma) 30 as described previously 31 . Raw data and all relevant data generated during analysis, including the used databases have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository 32 with the dataset identifier PXD005860.…”

Section: Methodsmentioning

confidence: 99%

Proteomic analysis of lung metastases in a murine breast cancer model reveals divergent influence of CTSB and CTSL overexpression

Sigloch¹,

Tholen²,

Gomez-Auli³

et al. 2017

J. Cancer

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Studies in the MMTV-PyMT (PyMT) breast cancer mouse model have shown a strong influence of the lysosomal cysteine cathepsins B or L on lung metastasis formation. Transgenic expression of human CTSB (tgCTSB) or CTSL (tgCTSL) both led to similar metastatic phenotypes with increased metastatic burden in the PyMT mice. However, recent studies in other tumor models proved marked differences in effects of either cathepsin on the proteome composition. We sought to analyze and compare proteome changes in the metastatic proteome of PyMT mice expressing either tgCTSB or tgCTSL to evaluate similarities and differences in those models.Performing an explorative, quantitative proteome comparison based on LC-MS/MS, we identified up to 3,000 proteins from murine lung metastases in three independent biological replicates per genotype. In both cases, when compared to wild-type (WT) mice, we noticed a pronounced impact of transgene cathepsin expression on the metastasis proteome. Highlights include increased moesin, integrin beta 1 and vinexin levels in the tgCTSB dataset and increased saposin and granulin levels in the tgCTSL dataset. Importantly, non-supervised hierarchical clustering clearly separated tgCTSB vs. tgCTSL induced proteome changes.In summary, tgCTSB and tgCTSL both display a strong and distinct impact on proteome composition of lung macrometastases in the PyMT model. Our observations suggest that they impact malignant behavior in distinct ways, thus further emphasizing interest into their tumor-contextual functionality.

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“…Proteins were only further considered if they were identified and quantified in at least four of the five patient cases for both comparisons. Files obtained by MaxQuant were further processed using RStudio v.0.99.446 (R Foundation for Statistical Computing, Vienna, Austria) as previously described [23] . Decoy sequences and potential contaminant entries were removed.…”

Section: Methodsmentioning

confidence: 99%

Proteomic Characterization of Prostate Cancer to Distinguish Nonmetastasizing and Metastasizing Primary Tumors and Lymph Node Metastases

et al. 2018

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Patients with metastatic prostate cancer (PCa) have a poorer prognosis than patients with organ-confined tumors. We strove to uncover the proteome signature of primary PCa and associated lymph node metastases (LNMs) in order to identify proteins that may indicate or potentially promote metastases formation. We performed a proteomic comparative profiling of PCa tissue from radical prostatectomy (RPE) of patients without nodal metastases or relapse at the time of surgical resection (n = 5) to PCa tissue from RPE of patients who suffered from nodal relapse (n = 5). For the latter group, we also included patient-matched tissue of the nodal metastases. All samples were formalin fixed and paraffin embedded. We identified and quantified more than 1200 proteins by liquid chromatography tandem mass spectrometry with subsequent label-free quantification. An increase of ribosomal or proteasomal proteins in LNM (compared to corresponding PCa) became apparent, while extracellular matrix components rather decreased. Immunohistochemistry (IHC) corroborated accumulation of poly-(ADP-ribose)-polymerase 1 and N-myc-downstream-regulated-gene 3, alpha/beta hydrolase domain-containing protein 11, and protein phosphatase slingshot homolog 3 in LNM. These findings strengthen the present interest in examining PARP inhibitors for the treatment of aggressive PCa. IHC also corroborated increased abundance of retinol dehydrogenase 11 in metastasized primary PCa compared to organ-confined PCa. Generally, metastasizing primary tumors were characterized by an enrichment of proteins involved in cellular lipid metabolic processes with concomitant decrease of cell adhesion proteins. This study highlights the usefulness of a combined proteomic-IHC approach to explore novel aspects in tumor biology. Our initial results open novel opportunities for follow-up studies.

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Impact of cathepsin B on the interstitial fluid proteome of murine breast cancers

Cited by 23 publications

References 54 publications

Non-invasive in vivo imaging of tumour-associated cathepsin B by a highly selective inhibitory DARPin

Non-invasive in vivo imaging of tumour-associated cathepsin B by a highly selective inhibitory DARPin

Proteomic analysis of lung metastases in a murine breast cancer model reveals divergent influence of CTSB and CTSL overexpression

Proteomic Characterization of Prostate Cancer to Distinguish Nonmetastasizing and Metastasizing Primary Tumors and Lymph Node Metastases

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