Abnormal Profile of Human Nucleolytic Activity as a Test for Cancer

Drake, Walter P.; Schmukler, Morton; Pendergrast, W. Jefferson; Davis, Alan; Lichtenfeld, J. Leonard; Mardiney, Michael R.

doi:10.1093/jnci/55.5.1055

Cited by 13 publications

(3 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In each study, control subjects were apparently normal and did not include patients with nonmalignant diseases. Correlation of the degree of increase of RNase activity with the extent of neoplasia or thera- peutic response was variable; five studies (8-10, 12, 14) found a correlation and two (11,12) (19), and allowed use of a higher pH, emphasizing possible contributions of pancreatic RNase with its higher pH optimum (20) to serum activity. Serum RNase activity is not "absolutely dependent" on phosphate or citrate as stated by Reddi (20).…”

Section: Discussionmentioning

confidence: 99%

Serum RNase in the diagnosis of pancreatic carcinoma.

Peterson¹

1979

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Serum RNase (RNase I; ribonucleate 3'-pyrimidino-oligonucleotidohydrolase, EC 3.1.4.22) activity (mean ± SD) with polycytidine as substrate was determined in normal individuals (24.9 ± 3.0 units/ml) and in patients with pancreatic cancer (37.3 + 14.8), pancreatitis (38.5 ± 12.6), nonpancreatic diseases (48.7 ± 14.8), or renal failure (175.8 : 92.8 Blood drawn from patients and controls was allowed to clot and then was centrifuged; serum was removed within 2 hr of collection. The serum was stored at -20'C. Freezing, thawing, and frozen storage for at least 1 month do not affect RNase activity.The assay for serum RNase activity was based on the method for bovine pancreatic RNase first reported by Zimmerman and Sandeen (2) with the synthetic substrate polycytidine. Serum was diluted 1:1000 with 5 mM Tris at pH 7 containing 0.01% bovine serum albumin, and a 50-sl aliquot was added to a Microfuge tube containing 150 Al of 0.1 M Tris at pH 8 and 50 /1l of polycytidine at 1 mg/ml. After incubation for 20 min at 370C, 250-A.l of cold 1.2 M perchloric acid/20 mM La203 was added. After 10 min at 00C, the mixture was centrifuged at 8000 X g for 5 min at room temperature. The supernatant, containing acid-soluble nucleotide, was diluted with water and the A280 was measured in a Beckman spectrophotometer. All serum dilutions and assays were performed in duplicate. Substrate blanks containing all reagents but no enzyme (serum) were analyzed in an identical manner and the values were subtracted from each assay. An enzyme (serum) blank proved unnecessary because serum did not contribute to the absorbance at these low dilutions. Sera assayed previously or bovine pancreatic RNase served as standards and their analysis was repeated with each group of sera of unknown activity. One RNase unit is the amount of enzyme activity that produces 1 ,mol of acid-soluble nucleotide per min at 37°C in 0.06 M Tris at pH 8. Molarity of acid-soluble nucleotide was calculated on the basis of an extinction coefficient of 13.0 X 103 at pH 2.0 (3).In 17 determinations on aliquots of one serum sample, performed consecutively, the mean (±SD) was 22.4 ± 0.7 units/ml. In 6 to 17 determinations on sera of four normal individuals obtained and performed at three different times, the mean was 21.3 ± 0.8 units/ml. Overall, 117 determinations were performed on the three samples of these four individuals; the results demonstrate remarkable precision of the procedure. As would be desired, the methodological variation is significantly less than the biologic variability: the mean of analyses on sera from 20 female and 20 male normal individuals was 24.9 + 3.0 units/ ml.The activity of bovine pancreatic RNase was the same when assayed with the polycytidine substrate used in my laboratory or with that used by Zimmerman and Sandeen (2) (Fig. 1A).When increasing amounts of three different sera were added to substrate, the rate of appearance of reaction products was linear (Fig. 1B).Patients were divided into five groups according to the following criteria. 263...

show abstract

Section: Discussionmentioning

confidence: 99%

Serum RNase in the diagnosis of pancreatic carcinoma.

Peterson¹

1979

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…Nucleases and their activity have been proposed as valuable prognostic markers for cancer. 52 , 53 , 54 , 55 , 56 , 57 , 58 For example, the expression of ERCC1, a DNA repair enzyme with nuclease activity, has been shown to be elevated in BCa CTCs that are resistant to chemotherapy. 59 Similarly, the expression of several other nuclease genes that belong to the class of DNA repair enzymes has also been shown to be elevated in cancer stem cells, which are thought to lead to metastasis.…”

Section: Discussionmentioning

confidence: 99%

Rapid and Sensitive Detection of Breast Cancer Cells in Patient Blood with Nuclease-Activated Probe Technology

Kruspe

Dickey

Urak

et al. 2017

Molecular Therapy - Nucleic Acids

View full text Add to dashboard Cite

A challenge for circulating tumor cell (CTC)-based diagnostics is the development of simple and inexpensive methods that reliably detect the diverse cells that make up CTCs. CTC-derived nucleases are one category of proteins that could be exploited to meet this challenge. Advantages of nucleases as CTC biomarkers include: (1) their elevated expression in many cancer cells, including cells implicated in metastasis that have undergone epithelial-to-mesenchymal transition; and (2) their enzymatic activity, which can be exploited for signal amplification in detection methods. Here, we describe a diagnostic assay based on quenched fluorescent nucleic acid probes that detect breast cancer CTCs via their nuclease activity. This assay exhibited robust performance in distinguishing breast cancer patients from healthy controls, and it is rapid, inexpensive, and easy to implement in most clinical labs. Given its broad applicability, this technology has the potential to have a substantive impact on the diagnosis and treatment of many cancers.

show abstract

“…Nucleases and their activity have been proposed as valuable prognostic markers for cancer. (269)(270)(271)(272)(273)(274)(275) For example, the expression of ERCC1, a DNA repair enzyme with nuclease activity, has been shown to be elevated in BCa CTCs that are resistant to chemotherapy. (276) Similarly, the expression of several other nuclease genes that belong to the class of DNA repair enzymes has also been shown to be elevated in cancer stem cells, which are thought to lead to metastasis.…”

Section: Discussionmentioning

confidence: 99%

Detection and treatment of critical illnesses using oligonucleotides

Urak¹

View full text Add to dashboard Cite

Sepsis is among the most prevalent diagnosed critical illnesses in the United States today. Although advances have reduced the overall morbidity and mortality associated with this illness, the enormous number of deaths associated with it shows a need for improved diagnostic and therapeutic options. Our laboratory has utilized RNA based technologies to aid in the treatment of histone induced multiple organ dysfunction syndrome seen in sepsis.Histones are proteins found in the nucleus of every cell in our body and have been shown to be released during sepsis. Such release induces damage to other cells, causing a feed forward cycle that results in organ failure and death. Several therapeutics have been utilized to neutralize histones but have shown considerable toxicity. This thesis describes the generation of single stranded RNA aptamers to bind and neutralize histone mediate damage without unwanted toxicity. We demonstrate that our aptamers selectively bind to histones but not serum proteins. In addition, we establish that our aptamers can neutralize all histone mediated cellular response in vitro and in vivo. Finally, we determined that our aptamers are able inhibit the histone feed forward cycle in a temporal fashion in our murine model of multiple organ dysfunction. This novel therapeutic demonstrates the selectivity and effectiveness needed to inhibit histones in several critical illnesses. iv PUBLIC ABSTRACTThe most prevalent diagnosed critical illness in the western hemisphere is sepsis.Despite recent advances in treatments, the number of deaths associated with sepsis continues to grow, resulting in the need for improved diagnostic and therapeutic options.Our laboratory has utilized RNA based technologies to aid in the treatment and detection of the critical illness stated above.During a critical illness such as sepsis, cells are damaged causing the release of proteins from the nucleus (i.e. histones) resulting in further cellular damage that leads to organ failure and death. To date, one FDA approved therapeutic has been utilized to neutralize histones but resulted in considerable toxicity in the patients. In this thesis we describe a novel therapeutic (single-stranded RNA aptamers) to bind and neutralize histone mediated damage without the toxic side-effects. This novel therapeutic demonstrates the selectivity and efficacy needed to inhibit histones in several critical illnesses.

show abstract

Abnormal Profile of Human Nucleolytic Activity as a Test for Cancer

Cited by 13 publications

References 0 publications

Serum RNase in the diagnosis of pancreatic carcinoma.

Serum RNase in the diagnosis of pancreatic carcinoma.

Rapid and Sensitive Detection of Breast Cancer Cells in Patient Blood with Nuclease-Activated Probe Technology

Detection and treatment of critical illnesses using oligonucleotides

Contact Info

Product

Resources

About