Evaluation of urine tumor-associated trypsin inhibitor, CYFRA 21-1, and urinary bladder cancer antigen for detection of high-grade bladder carcinoma.

OBJECTIVES: To assess the value of urine tumor-associated trypsin inhibitor (TATI), CYFRA 21-1, which measures cytokeratin 19 fragment, and urinary bladder carcinoma antigen (UBC) for the detection of high-grade bladder carcinoma. METHODS: A total of 160 individuals were enrolled in the present study. Of these, 80 were patients with proven primary high-grade urothelial bladder cancer (group 1), 40 were healthy volunteers (group 2), and 40 had history of benign urologic disease (group 3). All were evaluated with respect to urinary TATI, CYFRA 21-1, and UBC levels. All these markers were evaluated using commercial kits. Cytology was also performed. RESULTS: The TATI measurements were significant greater in group 1 compared with groups 2 and 3. The cutoff point used for TATI, CYFRA 21-1, and UBC was 22, 2.8, and 12 microg/L, respectively. The overall sensitivity was 85.7% for TATI, 61.9% for CYFRA 21-1, 50% for UBC, and 42.8% for cytology. TATI was significantly more sensitive in Stage Ta (80%) than was CYFRA 21-1 (32%), UBC (12%), and cytology (20%). TATI was also more sensitive compared with other tumor markers for Stage T1 but not for Stage T2 or T3. CONCLUSIONS: The results of our study have shown that TATI is a promising urinary tumor marker for high-grade urothelial bladder cancer. It is more sensitive than CYFRA 21-1, UBC, and cytology for Stage Ta and T1 bladder cancer.

Trypsin-2 degrades human type II collagen and is expressed and activated in mesenchymally transformed rheumatoid arthritis synovitis tissue.

It has traditionally been believed that only the human collagenases (matrix metalloproteinase-1, -8, and -13) are capable of initiating the degradation of collagens. Here, we show that human trypsin-2 is also capable of cleaving the triple helix of human cartilage collagen type II. We purified human trypsin-2 and tumor-associated trypsin inhibitor by affinity chromatography whereas collagen type II was purified from cartilage extracts using pepsin digestion and salt precipitation. Degradation of type II collagen and gelatin by trypsin-2 was demonstrated with sodium dodecyl sulfate-polyacrylamide gel electrophoresis, zymography, and mass spectrometry, and tumor-associated trypsin inhibitor specifically inhibited this degradation. Although human trypsin-2 efficiently digested type II collagen, bovine trypsin did not. Furthermore, immunohistochemical staining detected trypsin-2 in the fibroblast-like synovial lining and in stromal cells of human rheumatoid arthritis synovial membrane. These findings were confirmed by reverse transcriptase-polymerase chain reaction and nucleotide sequencing. Trypsin-2 alone and complexed with alpha(1)-proteinase inhibitor were also detected in the synovial fluid of affected joints by time-resolved immunofluorometric assay, suggesting that trypsin-2 is activated locally. These results are the first to assess the ability of human trypsin to cleave human type II collagen. Thus, trypsin-2 and its regulators should be further studied for use as markers of prognosis and disease activity in rheumatoid arthritis.

Urinary levels of tumor-associated trypsin inhibitor (TATI) in the detection of transitional cell carcinoma of the urinary bladder.

OBJECTIVE: To assess whether urinary levels of tumor-associated trypsin inhibitor (TATI) would aid in the detection of bladder transitional cell carcinoma (TCC); and to compare diagnostic performance of urinary TATI with that of nuclear matrix protein 22 (NMP22) and barbotage cytology. METHODS: We determined TATI and NMP22 levels in voided urine from 181 subjects: 153 with previous bladder cancer, 20 with urologic pathology other than bladder cancer, and eight healthy volunteers. TATI was analyzed continuously and categorically on the basis of its quartile distribution. We also measured urinary creatinine and barbotage cytology in 173 and 154 patients, respectively. RESULTS: Urinary TATI levels were significantly higher in TCC patients with evidence of tumor on cystoscopic evaluation (n = 96) than in control subjects (n = 85; p < 0.001). Higher levels of TATI were associated with positive cytology assay results (p = 0.018), higher NMP22 levels (p < 0.001), and invasive tumor stage (p = 0.026). The area under the receiver operating characteristics (ROC) curve (AUC) of TATI for the detection of TCC was 0.712 (95%CI: 0.637-0.786). The overall AUCs for TATI and NMP22 were not statistically different from each other (p = 0.174). In the >75% sensitivity region of the ROC curves, TATI was consistently more specific than NMP22. TATI, NMP22, and cytology were independently associated with bladder cancer (p = 0.049, p = 0.040, and p < 0.001, respectively). Adjustment of TATI for urinary creatinine levels did not affect any of the outcomes. CONCLUSIONS: Urinary level of TATI may add independent information to urinary cytology and NMP22 in the detection of bladder TCC. TATI seems to outperform NMP22 for bladder TCC detection.

Tumor-associated trypsin inhibitor as a prognostic factor during follow-up of bladder cancer.

OBJECTIVES: To evaluate the prognostic value of tumor-associated trypsin inhibitor (TATI) in the serum and urine of patients in follow-up for urinary bladder cancer. METHODS: Serum and urine samples were taken during follow-up of 157 patients with transitional cell carcinoma of the bladder who were monitored by cystoscopy and cytology in 1986 to 1987. Initially, 117 (75%) of the 157 tumors were superficial. At the time of sampling, 93 patients (59%) had no detectable tumor and 48 (31%) had a superficial, and 16 (10%) an invasive, tumor. Cancer-specific survival was evaluated in 1998. RESULTS: During follow-up, 35 patients (22%) died of bladder cancer. An elevated TATI concentration in the serum (21 microg/L or more) was associated with a significantly shorter survival (P <0.001) compared with a normal value. Multivariate analysis showed that serum TATI and detectable cancer at sampling were independent prognostic factors (P <0.001 and P = 0.002, respectively), and age, grade, urine cytology findings, and urine TATI were not. CONCLUSIONS: Serum TATI is an independent prognostic factor in transitional cell carcinoma and is potentially useful for the identification of patients with an adverse prognosis.

Identification of genes up-regulated in urothelial tumors: the 67-kd laminin receptor and tumor-associated trypsin inhibitor.

Studies investigating changes in gene expression in urothelial carcinoma have generally compared tumors of different stages and grades but comparisons between low-grade, noninvasive tumors and normal urothelium are needed to identify genes involved in early tumor development. We isolated the urothelium from a low-grade tumor and corresponding normal mucosa by laser capture microdissection on frozen sections. The RNA extracted was amplified to generate suppressive subtractive cDNA libraries. Random sequencing of cDNA clones identified approximately 100 unique species. Of these 83% were known genes, 15% had homology to genes with an unknown function in humans, and 2% did not show homology to any published gene sequence. Two of the known genes, the 67-kd laminin receptor (67LR) and tumor-associated trypsin inhibitor (TATI), had previously been associated with metastatic progression in many tumor types, although 67LR has not been investigated in urothelial tumors. Immunolabeling of the original tissue with antibodies against these two genes confirmed overexpression, validating our strategy: 67LR was not expressed in the normal urothelium but was present in the tumor, whereas TATI expression was confined to umbrella cells in the normal urothelium, but extended to all cell layers in the tumor. We investigated both markers further in a separate series of tumors of different stages and grades. TATI was more consistently overexpressed than 67LR in all tumor grades and stages. Levels of secreted TATI were significantly higher in urine samples from patients with tumors compared to controls. Our strategy, combining laser capture microdissection and cDNA library construction, has identified genes that may be involved in the early phases of urothelial tumor development rather than with disease progression, highlighting the importance of comparing tumor with normal rather than just tumors of different stages and grades.

Tumor-associated trypsin inhibitor (TATI) and renal function.

Tumor-associated trypsin inhibitor (TATI) is a low molecular weight protein employed as tumor marker. To evaluate the role of the kidney in the clearance of TATI we studied the relationship of serum TATI with the glomerular filtration rate (GFR), and for comparisons the relationships of beta 2-microglobulin (beta(2m)) and creatinine with GFR. Urine excretion and renal extraction of TATI were also determined. The decrease in GFR was accompanied by an increase in blood levels of TATI, beta(2m) and creatinine. Serum TATI increased 12.4 times in patients with renal failure (GFR < 20 ml/min) with respect to subjects with normal renal function (P < 0.001, non-parametric Mann-Whitney test), while beta(2m) increased 7.3 times (P < 0.001) and creatinine 4.7 times (P < 0.001). In patients with GFR 60 to 40 ml/min, only the increase in TATI was statistically significant (p < 0.005). Renal excretion of TATI was low but it increased progressively in renal failure. Renal extraction ranged from 13% to 41%, for a mean 24.87. These results suggest that TATI is handled by the kidney and that it is a snesitrive marker of reduction in renal function.

Tumour-associated trypsin inhibitor (TATI) in ovarian cancer.

Tumour-associated trypsin inhibitor (TATI) is a 6 kD peptide isolated from the urine of a patient with ovarian cancer. Increased urinary excretion of TATI has earlier been observed in patients with gynaecological malignancies. The value of TATI in urine and serum as a marker for ovarian cancer was studied in 102 patients. Preoperatively urine TATI was elevated in 55% (18/33) and serum TATI in 27% (12/45) of the patients. In patients with mucinous tumours, elevated preoperative levels of TATI were observed in 6 out of 10 patients, while CA 125 was elevated in 4 and CEA in one of the cases. When assay of TATI was used to predict presence of disease before second-look surgery of 48 patients, the sensitivity and specificity of serum TATI was 19% and 91%, and that of urine TATI 42% and 76%, respectively. Rising TATI levels were observed in progressive disease, whereas regressive disease was not as often associated with falling levels. Serum TATI was elevated in 45% (144/318) and urine TATI in 57% (73/171) of samples from patients with clinical evidence of disease. The TATI assay was found to be of potential value in the management of patients with mucinous ovarian cancer, but in patients with non-mucinous ovarian cancer it did not provide information additional to that obtained from assay of ovarian cancer marker CA 125 alone.

Pancreatic secretory trypsin inhibitor from human amniotic fluid and fetal and neonatal urine: concentrations and physicochemical characterization.

The levels and physicochemical properties of the pancreatic secretory trypsin inhibitor, also known as Kazal type trypsin inhibitor, were studied in human amniotic fluid. In the second trimester, the median concentration was 160 ng/ml, which exceeds the maternal serum levels 20-fold. Towards term, the amniotic fluid levels declined about 5-fold, whereas the maternal serum values remained constant. In fetal urine, the concentration of the trypsin inhibitor was similar to that in amniotic fluid in early gestation, whereas in newborn urine, the median level was 4-to 5-fold higher than in term amniotic fluid. The physiochemical characteristics of the trypsin inhibitor in amniotic fluid, neonatal urine and cancer urine from an ovarian cancer patient were similar, as studied by gel filtration, high performance reverse phase liquid chromatography, and complete immunological identity in immunodiffusion. The physicochemical similarity and levels in various compartments suggest fetal contribution to amniotic fluid levels of the trypsin inhibitor.

Pancreatic secretory trypsin inhibitor-like immunoreactivity in pancreatectomized patients.

Pancreatic secretory trypsin inhibitor (PSTI) is a 6000-dalton peptide, that occurs in high concentrations in the pancreas and in pancreatic juice. It is thought to be synthesized by pancreatic acinar cells. We have recently reported the findings of an identical trypsin inhibitor at high concentrations in the urine of patients with gynecological malignancy. Therefore, we have named the inhibitor tumor-associated trypsin inhibitor (TATI). We have now studied patients who have undergone total pancreatoduodenectomy for pancreatic cancer or chronic pancreatitis. By radioimmunoassay (RIA), we found normal levels of this inhibitor in the serum and urine of pancreatectomized patients. The absence of pancreas was confirmed by measuring serum trypsin. By gel filtration and HPLC it was found that PSTI/TATI occurring in pancreatectomized patients was indistinguishable from that found in connection with pancreatitis and ovarian cancer.

Elimination of pancreatic secretory trypsin inhibitor from the circulation. A study in man.

The elimination of human pancreatic secretory trypsin inhibitor (PSTI) from the circulation of man has been examined in two human volunteers. Serial samplings of blood and urine were made for 55 h, after a rapid intravenous infusion of 125I-labeled human PSTI. The findings demonstrated a rapid initial elimination from the circulation. Within 30 min only 30% of the infused label remained (T1/2, 6 min). This was accompanied by the rapid appearance in the urine of radiolabel. Our results indicate that PSTI would prove a poor diagnostic marker for acute pancreatitis late in the course of the disease. This is opposed to the findings of Ogawa et al., who reported prolonged elevated circulating levels of PSTI weeks into the disease. However, they also noted rises of PSTI during acute pancreatitis in excess of 10 times the levels we have noted. Further exploration of population differences and the behavior of PSTI during acute pancreatitis are necessary to help resolve these findings.

Excretion of a tumor-associated trypsin inhibitor (TATI) in urine of patients with gynecological malignancy.

In earlier studies we reported the finding of a tumor-associated peptide that also occurred at high concentrations in early amniotic fluid. Determination of the N-terminal sequence of this peptide revealed that it is closely related or identical to the pancreatic secretory trypsin inhibitor. Therefore, the peptide is called tumor-associated trypsin inhibitor (TATI). The concentration of TATI was determined by radioimmunoassay in the urine of 148 patients with various forms of gynecologic malignancy and in a reference population consisting of 98 patients with non-malignant gynecologic disease, and also in 40 patients with severe infections or inflammatory disease. In the reference population, the median urinary concentration of TATI was 22 micrograms/g creatinine and the central 95% reference interval was 7-50 micrograms/g creatinine. Elevated urinary levels were observed in 53% of all patients with gynecologic cancer, in 63% of those with active disease and 26% of those in clinical remission. The highest urinary TATI level (11,000 micrograms/g creatinine) was over 200 times the upper limit of the reference range. Patients with cervical cancer had the highest frequency of elevated values. Increased excretion of TATI was also observed in patients with severe bronchopulmonary infections and pancreatitis. Although increased excretion of TATI is not cancer-specific, the distinction by elevated levels of TATI between malignant and nonmalignant gynecologic disease is better than by most other putative tumor markers, and the increased excretion of TATI in patients with active disease can be important for the understanding of tumor biology.