Polymer-Ligand-Based ELISA for Robust, High-Throughput, Quantitative Detection of p53 Aggregates

Maritschnegg, Elisabeth; Heinzl, Nicole; Wilson, Stuart M.; Deycmar, Simon; Niebuhr, Markus; Klameth, Lukas; Holzer, Barbara; Koziel, Katarzyna; Concin, Nicole; Zeillinger, Robert

doi:10.1021/acs.analchem.8b02373

Cited by 20 publications

(15 citation statements)

References 33 publications

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“…To quantify the level of PTEN aggregation in these samples, we employed the so-called Seprion-ELISA, which was previously shown to yield highly reliable quantification of amyloid like aggregation in tissues (Maritschnegg et al, 2018). Similar to what was observed in vitro and in PDTX models, only 3/20 (15%) of type I tumors showed an absorbance above the threshold in the Seprion-ELISA, whereas background levels were measured in 17/20 (85%).…”

Section: Exploratory Retrospective Study Of Pten Aggregation In Endommentioning

confidence: 73%

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The tumor suppressor protein PTEN undergoes amyloid-like aggregation in tumor cells

Claes

Maritschnegg

Baets

et al. 2020

Preprint

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Protein aggregation is an underappreciated mechanism that may contribute to the loss- and oncogenic-gain-of-function of mutant tumor suppressors such as p53 and axin. In the present study, we describe amyloid-like aggregation behaviour of the second most frequently mutated tumor suppressor in human cancer, PTEN. In silico analysis revealed a particularly high aggregation vulnerability for this protein, which was corroborated by in vitro aggregation assays. In cultured tumor cells, we found that under stress conditions, PTEN readily undergoes amyloid-like aggregation as a result of mutation. However, we also show that severe dysregulation of protein homeostasis may lead to aggregation of wild-type PTEN. These observations were supported by a small survey of patient-derived uterine tumor tissues, which found that more than 25% of tumors analyzed displayed wild-type PTEN aggregation. Finally, in an exploratory clinical study we found that PTEN aggregation status was correlated with a decline in clinical outcome. Our findings establish that the tumor suppressor PTEN is highly aggregation-prone and our work suggests that protein aggregation might be an underestimated but prevalent component of cancer cell biology.

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Section: Exploratory Retrospective Study Of Pten Aggregation In Endommentioning

confidence: 73%

“…The PTEN aggregation load was determined using the Seprion-ELISA (Microsens) as described in (Maritschnegg et al, 2018). This test is based on a polyionic, high-molecular-weight ligand, coated to the surface of the ELISA plate.…”

Section: Seprion-elisamentioning

confidence: 99%

The tumor suppressor protein PTEN undergoes amyloid-like aggregation in tumor cells

Claes

Maritschnegg

Baets

et al. 2020

Preprint

Self Cite

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“…Following the finding that p53 associates into amyloid-like aggregates, an ELISA system based on a polyionic, highmolecular-weight ligand that specifically captures aggregated oligomers and amyloid proteins was developed. Naturally occurring tetramers of the p53 are not bound, but high-molecular-weight aggregates are detected in the ovarian cancer cell lines and patientderived tumor tissues [284]. Such aggregates could be potential targets for the development of novel therapeutic strategies against tumors.…”

Section: Crosstalk Between P53/p73 Isoformsmentioning

confidence: 99%

p53/p73 Protein Network in Colorectal Cancer and Other Human Malignancies

Horvat

Tadijan

Vlašić

et al. 2021

Cancers

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The p53 tumor suppressor protein is crucial for cell growth control and the maintenance of genomic stability. Later discovered, p63 and p73 share structural and functional similarity with p53. To understand the p53 pathways more profoundly, all family members should be considered. Each family member possesses two promoters and alternative translation initiation sites, and they undergo alternative splicing, generating multiple isoforms. The resulting isoforms have important roles in carcinogenesis, while their expression is dysregulated in several human tumors including colorectal carcinoma, which makes them potential targets in cancer treatment. Their activities arise, at least in part, from the ability to form tetramers that bind to specific DNA sequences and activate the transcription of target genes. In this review, we summarize the current understanding of the biological activities and regulation of the p53/p73 isoforms, highlighting their role in colorectal tumorigenesis. The analysis of the expression patterns of the p53/p73 isoforms in human cancers provides an important step in the improvement of cancer therapy. Furthermore, the interactions among the p53 family members which could modulate normal functions of the canonical p53 in tumor tissue are described. Lastly, we emphasize the importance of clinical studies to assess the significance of combining the deregulation of different members of the p53 family to define the outcome of the disease.

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“…Abnormal expression of MTp53 (mutant-type p53 protein) can serve as an important carcinogenesis stimulus [21]. Thus far, various methods for detecting endogenous p53 have been reported, including enzyme-linked immunosorbent assay (ELISA) [22], immunohistochemically assay [23], electrophoretic mobility shift assay [24], and ds-DNA consensus binding biosensing assay [25]. Despite the favorable detection limit and sensitivity of these methods, none of these methods linked at the relationship between photothermal therapy and p53 protein signaling pathways.…”

Section: Introductionmentioning

confidence: 99%

A Gold Nanoparticle Bouquet held on plasma membrane: An ultrasensitive dark-field imaging approach for Cancer Cell Analysis

et al. 2020

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Rational: p53 is suppressing tumor protein correlated with the cell cycle factors and apoptosis. Here, a gold nanoparticle bouquet is designed for an ultrasensitive dark-field imaging approach for cancer cell analysis. Methods: AuNP60/APBA is functionalized by a gold nanoparticle bouquet-plasmonic 60 nm gold nanoparticles. And consistent APBA can be held on the plasma membrane. After 13 nm gold nanoparticles are functionalized with mannose (AuNP13/MN), the AuNP60/APBA gold nanoparticles are captured. The absorption spectrum of aggregation gold nanoparticles (AuNPs) shifts to near-infrared (NIR) region which can be observed under dark-field microscopy (DFM) and is treated the subsequent with photothermal therapy. Results:The results that MCF-7 cells were successfully destroyed under the near-infrared (NIR) irradiation and the intracellular WTp53 increased while the MTp53 decreased. These results indicated that p53 is the key molecule in the apoptosis signaling pathway. Photothermal therapy can stimulate the MTp53 in the cell signal conductive pathway. Conclusion: This work offers a new method for intracellular p53 analysis and a potential targeted cancer treatment.

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Polymer-Ligand-Based ELISA for Robust, High-Throughput, Quantitative Detection of p53 Aggregates

Cited by 20 publications

References 33 publications

The tumor suppressor protein PTEN undergoes amyloid-like aggregation in tumor cells

The tumor suppressor protein PTEN undergoes amyloid-like aggregation in tumor cells

p53/p73 Protein Network in Colorectal Cancer and Other Human Malignancies

A Gold Nanoparticle Bouquet held on plasma membrane: An ultrasensitive dark-field imaging approach for Cancer Cell Analysis

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