Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Balian, A.; González, Javier García; Bastida, Nora; Akhtar, Khadija-Tul Kubra; Borsa, Barış Ata; Hernandez, Frank J.

doi:10.3791/60005

Cited by 6 publications

(11 citation statements)

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“…More specifically, readily available chemically modified nucleosides enable the design and development of more tailored oligonucleotide libraries with enhanced sensitivity and specificity towards the target nuclease. The concept of screening and probe design in an iterative manner to yield best performing probes has been comprehensively described in [86]. Worth mentioning is that nuclease activity has been exploited as a biomarker for the detection of cancer cells by an independent research group, where researchers have been able to detect circulating tumor cells enriched from breast cancer patients based on the cells' nuclease activity [87].…”

Section: Serum Rnase Activitymentioning

confidence: 99%

“…While most of the studies focus on comparing the expression status of a specific nuclease, at protein or mRNA level, for a cancer type, less effort has been dedicated to interrogating the dynamic nucleolytic activity of these enzymes as a potential cancer biomarker. We and others have been among the leading research groups that properly discussed and developed this notion [84,86,87]. In fact, the reported difference in nuclease profile as protein or mRNA between cancer and normal has led Hernandez et al to anticipate that nuclease activity signature could be recruited as a diagnostic biomarker taking advantage of the dynamic property of these enzymes.…”

Section: Advantages Potential and Future Directions Of Nuclease Activity As A Biomarker In Cancer Diagnosismentioning

confidence: 99%

“…Implementation of nuclease activity as a biomarker tackles the accuracy problem through developing nucleic acid probes with enhanced sensitivity and specificity in an iterative manner [86]. In that, the probes' design is improved based on the sequence of candidate oligonucleotides from each screening round.…”

Section: Advantages Potential and Future Directions Of Nuclease Activity As A Biomarker In Cancer Diagnosismentioning

confidence: 99%

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Nucleases as molecular targets for cancer diagnosis

Balian

Hernandez

2021

Biomark Res

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Early cancer diagnosis is a crucial element to improved treatment options and survival. Great research efforts have been made in the search for better performing cancer diagnostic biomarkers. However, the quest continues as novel biomarkers with high accuracy for an early diagnosis remain an unmet clinical need. Nucleases, which are enzymes capable of cleaving nucleic acids, have been long considered as potential cancer biomarkers. The implications of nucleases are key for biological functions, their presence in different cellular counterparts and catalytic activity led the enthusiasm towards investigating the role of nucleases as promising cancer biomarkers. However, the most essential feature of these proteins, which is their enzymatic activity, has not been fully exploited. This review discusses nucleases interrogated as cancer biomarkers, providing a glimpse of their physiological roles. Moreover, it highlights the potential of harnessing the enzymatic activity of cancer-associated nucleases as a novel diagnostic biomarker using nucleic acid probes as substrates.

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Section: Serum Rnase Activitymentioning

confidence: 99%

Section: Advantages Potential and Future Directions Of Nuclease Activity As A Biomarker In Cancer Diagnosismentioning

confidence: 99%

Section: Advantages Potential and Future Directions Of Nuclease Activity As A Biomarker In Cancer Diagnosismentioning

confidence: 99%

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Nucleases as molecular targets for cancer diagnosis

Balian

Hernandez

2021

Biomark Res

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“…The bacterial culture is estimated to be in the stationary phase with values greater than 10 9 CFU/ mL after this incubation time of 24 h. Next, the culture was centrifuged at 4500 × g for 30 min and supernatants were collected for direct use or storage at 4 °C or − 20 °C. TSB culture medium was used as control [34]. Bacterial counts were calculated using McFarland standards and reported as colony forming units (CFU) per milliliter.…”

Section: Preparation Of Bacterial Supernatantmentioning

confidence: 99%

Activatable MRI probes for the specific detection of bacteria

Prabu

Balian

et al. 2021

Anal Bioanal Chem

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Activatable fluorescent probes have been successfully used as molecular tools for biomedical research in the last decades. Fluorescent probes allow the detection of molecular events, providing an extraordinary platform for protein and cellular research. Nevertheless, most of the fluorescent probes reported are susceptible to interferences from endogenous fluorescence (background signal) and limited tissue penetration is expected. These drawbacks prevent the use of fluorescent tracers in the clinical setting. To overcome the limitation of fluorescent probes, we and others have developed activatable magnetic resonance probes. Herein, we report for the first time, an oligonucleotide-based probe with the capability to detect bacteria using magnetic resonance imaging (MRI). The activatable MRI probe consists of a specific oligonucleotide that targets micrococcal nuclease (MN), a nuclease derived from Staphylococcus aureus. The oligonucleotide is flanked by a superparamagnetic iron oxide nanoparticle (SPION) at one end, and by a dendron functionalized with several gadolinium complexes as enhancers, at the other end. Therefore, only upon recognition of the MRI probe by the specific bacteria is the probe activated and the MRI signal can be detected. This approach may be widely applied to detect bacterial infections or other human conditions with the potential to be translated into the clinic as an activatable contrast agent.

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“…The availability of several chemically modified nucleotides [ 40 ] offers the possibility of tailoring oligonucleotide substrates for targeting nuclease activity derived from cancer, in a specific and sensitive manner [ 41 ]. Therefore, we hypothesized that the wide diversity of nucleases, together with their altered expression and deregulated activity in cancer, can be exploited as a new type of biomarker for the diagnosis, prognosis and possibly prediction of treatment response in many cancers, by using short and specific nucleic acids substrates.…”

Section: Introductionmentioning

confidence: 99%

Discovery and Proof-of-Concept Study of Nuclease Activity as a Novel Biomarker for Breast Cancer Tumors

Hernandez

Araúzo-Bravo

Gerovska

et al. 2021

Cancers

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Breast cancer is one of the most common pathologies diagnosed in the clinical practice. Despite major advancements in diagnostic approaches, there is no widely accepted biomarker in the clinical practice that can diagnose breast malignancy. Confirmatory diagnosis still relies on the pathological assessment of tissue biopsies by expert pathologists. Thus, there is an unmet need for new types of biomarkers and novel platform technologies that can be easily and robustly integrated into the clinic and that can assist pathologists. Herein, we show that nuclease activity associated to malignant tumors can be used as a novel biomarker in breast cancer, which can be detected via specific degradation of nucleic acid probes. In this study we have identified a set of three chemically modified nucleic acid probes that can diagnose malignancy in biopsy samples with high accuracy (89%), sensitivity (82%) and specificity (94%). This work represents a breakthrough for the potential clinical use of nuclease activity as biomarker, which can be detected via nucleic acids probes, for the clinical diagnosis of malignancy in breast tissue biopsies. This platform technology could be readily implemented into the clinic as adjunct to histopathological diagnostic.

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Kinetic Screening of Nuclease Activity using Nucleic Acid Probes

Cited by 6 publications

References 0 publications

Nucleases as molecular targets for cancer diagnosis

Nucleases as molecular targets for cancer diagnosis

Activatable MRI probes for the specific detection of bacteria

Discovery and Proof-of-Concept Study of Nuclease Activity as a Novel Biomarker for Breast Cancer Tumors

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