Abstract
Risk for colorectal cancer (CRC) confers susceptibility to other cancers such as small bowel, breast, prostate, and female genital. However, these cancer family histories (F/H) have not been ...studied systematically.
Methods: 1,948 patients at increased CRC risk were followed to determine if the biomarker outcome was related to personal cancer prevalence or a cancer F/H by risk questionnaire which categorized patients as: Group 0 (no family history of cancer), Group 1 (F/H of breast cancer), Group 2 (F/H CRC), and Group 3 (P/H uterine/ovarian cancers).
Results: Demographics (age, sex, race, obesity) and NSAID use were similar in all groups and 88% were males. The proportion of cancer incidence in Group 1 is statistically greater RR 1.36 (1.08-1.72) p = 0.014 compared to group 0 (Figure). Conversely, group 2 had statistically less cancer RR 0.72 0.52-0.99, p=0.045. There was no significance difference between Group 0 and Group 3. About half the tumors in this older group of Veterans were prostatic. There was a wide variety of other cancers. In the entire cohort of the 5 tumors the most common tumor was prostate (n=222) followed by lung (n=61); CRC (n=53); bladder (n=23) and breast (n=20).
Conclusions: These differences may be explained by the effect of exosomes which play a major role in CRC and other cancers. Exosomes are nano-lipid particles of 30-100 nm released into the extracellular compartment. The ability of exosomes to facilitate cell-cell communication could be used to inhibit tumorigenic pathways. Exosomes may be endogenous in the case of CRC cell release or exogenous, found in breast milk containing microRNA moieties relevant to inhibition of carcinogenesis of cancers. Many of these mutated genes may inhibit tumorigenic process of cancers thus conferring protection against incipient cancers thus explaining our observation in Group 2. A commonality between CRC and endometrial cancer genetics potentially explains the reduced cancer prevalence in Groups 2 & 3. Breast cancer does not share BRCA “founder genes” with CRC and in contrast to CRC, K-ras mutations are not major events and thus F/H does not confer protection. This hypothesis remains to be proven but in the interim, timely screening for common cancers in patients with a family history of breast cancer may be effective.
Citation Format: Martin Tobi, Fadi Antaki, MaryAnn Rambus, Michael Lawson, Daniel Ezekwudo. A family history of CRC confers protection against other cancers in a cohort of patients at risk for CRC abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1246.
Abstract
Current screening for lung cancer (LC) reduces mortality but tests are not ideal. Low-dose CT scans are expensive, incur radiation exposure, and high false positive rates. Although sputum ...cytology is unhelpful, saliva biomarker testing is promising but no prospective data are available.
Methods: In conducting a study using stool markers in a high risk colorectal neoplasia population we prospectively collected saliva and stool samples. Subsequently, 11 with available saliva; 8 with stool specimens but without significant personal or family history of GI neoplasia or symptoms, had contracted LC. We used 3 monoclonal antibodies to test reactivity in saliva (Adnab-9, BAC 18.1, COX-2). These are biomarkers of innate immune system, cell-mediated immunity, and inflammation, respectively. We used ELISA with either alkaline phosphatase (ALP) or immunoperoxidase (IMP) substrate and contrasted reactivity in LC patients and asymptomatic controls with no advanced polyps, and chemiluminescent dot blots with manual or Bel-blotter 96-well replicating tool.
Results: Saliva IMP testing was positive in 73% of 11 LC patients and 50% of 8 controls contrasted with ALP ELISA for stool Adnab-9 in 75% of 8 LC patients and 32% in 34 controls (OR 6.27:CI1.09-36.25;p<0.05).Specificity was 68%. Sensitivity for Adnab-9 for IMP manual and bel-blotting was 73 and 9% respectively; specificities were 37 and 64% respectively. BAC 18.1 sensitivities were 73 and 55% respectively; specificities 13 and 29%. COX-2 sensitivity for bel-blotting only was 27% and specificity was 71%. Inherent salivary peroxidase activity (OD<0.1/1μg protein) was negative in all 7 LC versus 9 of 23 (39%) of non-LC patients (p=0.07). The peroxidase absorbance meansSD were significantly different (0.0770.014 versus (0.1160.056;p<0.007). Equivalent inherent alkaline phosphatase of saliva samples was negative in both groups and means were not significantly different. The approximate time from saliva collection to diagnosis of LC was 3.76 years and 3.89 for stool.
Conclusions: Adnab-9 sensitivity was moderate but promising due to the ability to make an early preclinical diagnosis. While this was only significantly different from controls in stool ALP ELISA, inherent IMP activity could be blocked to improve specificity. Significantly suppressed inherent peroxidase activity in LC saliva may explain the insensitivity of the Bac18.1 and Cox2 inflammatory biomarkers. The Bel-blotter volume capacity is 4-10μl/blot and may explain the lower sensitivity using this tool. A battery of tests, including Adnab-9 in an ALP ELISA format may allow for early disease intervention.
Citation Format: Yosef Y. Tobi, Fadi Antaki, MaryAnn Rambus, Martin Tobi. Prospective lung cancer diagnostic screening using whole, unstimulated saliva versus stool abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 727. doi:10.1158/1538-7445.AM2017-727
Abstract
Introduction: Adenoma formation data in diabetics is inconclusive. Alcohol intake (AI), associated with colorectal neoplasia (CRN) has not been systematically studied in diabetes. We ...investigated effects of diabetes and AI in a prospective patient cohort at increased CRN risk.
Methods: Diabetes diagnoses based on patients' problem list, corroborated by HbA1c and glucose levels. Self-reported AI was quantified as more than occasional/social. Cumulative adenoma number were computed using available records. In a subset of patients, CRN-related Paneth cell marker p87 was used to semi-quantify immunohistochemically the colonic regions binding this label.
Results: 680 patients (348 diabetics 332 non-diabetic) were ascertained with 12.4-year median follow-up. Diabetics had more adenomas than non-diabetics (2.3±3.54 versus 1.81±2.91, p=0.05) and p87 levels were significantly higher in the descending colon (p<0.03) and rectum (p<0.008). Significant demographic differences of increased black ethnicity, lesser survival, increased BMI and increased smoking were appreciated in diabetics. In non-drinking patients, adenomas were also similarly increased in diabetics and demographics and p87 levels were as reported above without significant smoking differences. Comparing diabetics who drank actively with non-diabetics who did not, the diabetics had increased polyp accumulation (2.90±4.29 versus 1.62±2.45 p<0.012), increased proportion of black ethnicity, increased BMI and were younger. Rectal p87 was significantly increased (p<0.04) without smoking effects on polyp number.
Conclusion: Diabetics accumulate more adenomas than controls which is reflected in the increase of left-sided p87 expression. A combination of increased AI and diabetes particularly increases adenoma formation seemingly independent of factors such as age and smoking.
Citation Format: Martin Tobi, Fadi Antaki, Stephanie Judd, Paula Sochacki, MaryAnn Rambus, Harvinder Talwar, Michael Lawson, Edi Levi, Benita McVicker. Does a combination of type 2 diabetes mellitus and increased alcohol intake affect prospective total adenoma accumulation abstract. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2365.
Abstract
Lung cancer is the biggest cancer killer Worldwide with a limited prognosis despite personalized treatment regimens. Low-dose CT scanning as a means to early diagnosis has been disappointing ...due to the high false positive rate. Other non-invasive means of testing need to be developed that offer both timely diagnosis and predict prognosis. Methods: In the course of stool testing in a large scale testing of patients at increased risk of CRC we were able to ascertain 55 patients documented to have prospectively been diagnosed with lung cancer. Stool and colonic effluents were tested for anti-adenoma antibody (Adnab-9) reactivity by ELISA and Western blot. Survival data were obtained where available. Results: Of 55 cancers 40% were squamous (SSC), 23.7 were adenocarcinoma, 14.6 small, 12.7 large cell and 9% were designated NSCLC. 49.1% of the lung cancer patients had adnab-9 testing. Overall 53.6% (14 of 26) had positive testing compared to 2 of 11 controls (18.2%) which bordered on significance (OR5(CI1-29)). Cancers with higher lethality were more likely to test positive (67% for both small and large cell lung cancers, 50% SCC and adenocarcinoma, 33% NSCLC). In the larger groups overall survival was worse in those testing positive (474 versus 844 days in SCC and 54 versus 749 days in adenocarcinoma patients). Most importantly the time from a positive test to the clinical diagnosis ranged from 2.72 years for small cell, 3.13 for adenocarcinoma, 5.07 for NSCLC, 6.07 for SSC, and 6.24 for large cell cancer). In excluded cases where lung cancer was believed to be metastatic 75% of cancers were positive. Conclusions: At a projected real world sensitivity of 0.53 and specificity of 0.82 and the ability to predate diagnosis by up to 4.7 years overall, this test could help direct lung cancer screening. In addition, the Adnab-9 testing selectively detects worse tumor types (67%) and those with worse prognosis amongst the more common, favorable phenotypes thus making early diagnosis possible in those patients who stand to benefit most from this strategy. Metastatic lung cancer also detected by the test should be identified by the follow-up imaging studies and therefore would not be considered to be a major pitfall.
Citation Format: Martin Tobi, Yosef Y. Tobi, Fadi Antaki, MaryAnn Rambus, Michael Lawson. Preliminary data of a stool test for lung cancer from a prospective study abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4519.
Abstract
Colorectal cancer incidence and mortality impacts African Americans more than other ethnic group. While causes for this disparity are likely multifactorial, etiology of carcinogenesis may be ...the most important. A better understanding of these factors might enable us to design interventions based on regional realities. In the context of screening trials we obtained data from a cohort of patients with high-grade dysplastic adenomas. We compared differences between those with large-sized (>1cm) and those with smaller lesions. Methods: Patients were selected on the basis of high CRC risk based on a brief questionnaire. Prior to colonoscopy, stool was submitted for FOBT and anti-adenoma antibody (Adnab-9) binding determined by ELISA. The colonoscopy outcomes were noted for the cross-sectional element of the study. The longitudinal component enabled collection of additional specimens to elucidate the source of the adenoma antigen. Pathology was reviewed and antigenic mapping of 6 colonic segments was performed by Adnab-9 immunohistochemistry. If any 2 contiguous colonic segments expressed the antigen the colon was deemed to have a field effect. Results: 49 Patients were found to have HGD lesions of whom 26 were AA (53%). All but 1 were male. Mean age±SD was 62±14.8 years. 28 (57%) had LHiGDA and the remainder ShiGDA. 69% AA had LHiGDA versus 29% in the ShiGDA group (OR 5.9 CI 1.7-20.9;p = 0.03). Intragroup age differences between the AA and non-AA patients were not statistically (NSS) different (66.3±8.3 versus 63.9±10.5 and 58.4±21.9 versus 61.07±9.9 years respectively). Intergroup age variation by race was NSS (AA p = 0.2 and non-AA p = 0.5). Likewise intergroup lesion size variations were NSS (p = 0.4 and p = 0.2 respectively). The proportions of TVA were also NSS between the groups (p = 1 and p = 0.2 respectively}. The stool Adnab-9 binding was also not significantly different (p = 0.5 and p = 0.9 respectively). There was however a significant difference between the 2 lesion groups in terms of the presence of a field effect which was performed in a small percentage (14.3 versus 33%) for the large versus the small lesions (100 vs 14.3% RR7 CI 1.1-43;p<0.02) but numbers were too small for intragroup calculations. Conclusions: AA are overrepresented in the LHiGDA group but there were no clues for differing pathways of carcinogenesis or for the differential dwell times of these lesions. We did not evaluate habits such as alcohol intake and smoking as risk factors which conceivably may play a role. The morphology of the lesion and pathway antigens suggest that the adenoma-carcinoma is the pathway of causation but these should be further studied to design effective intervention and screening. Screening is likely to better detect the large than the small lesions and this may be an advantage for the AA population who accordingly should be encouraged to actively participate in local screening programs.
Citation Format: Martin Tobi, Fadi Antaki, MaryAnn Rambus, Paula Sochacki, Edi Levi. Detroit African Americans (AA) are overrepresented among patients with large high-grade dysplastic (HGD) adenomas (LHiGDA) versus those with small HGD adenomas (ShiGDA). abstract. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3714. doi:10.1158/1538-7445.AM2015-3714
Abstract
As a tumorigenesis model, colorectal cancer is associated with multiple gene mutations accumulating progressively but also has mutations that may alter disease course and provide a ...therapeutic target. Good examples of this are EGFR and VEGF. In melanoma, PD-1 interactions involving the immunobiome are therapeutically important. We conducted a study to detect potential mutations that might enhance therapy in colorectal cancer guided by expression of p87, a product of innate immune system Paneth cells. Methods: Adnab-9 immunohistochemistry or ELISA was used to define significant p87 Adenoma-associated antigen field effects (FE) in 10 patients with >1cm large high grade dysplastic adenomas (LHiGDA) and 3 with smaller high grade dysplastic adenomas (SHiGDA). We postulated that SHiGDA are not immunologically recognized by host defenses leading to negative outcomes. We used Ion Torrent™ sequencing (ITS) to find mutations in DNA (QiaAmp kit) extracted from 4 normal-appearing colonic segments taken from 1 patient in each group. Novel mutations found on ITS would then be sought using PCR with appropriate primers and subsequent sequencing of the PCR product circulating DNA extracted from available serum samples. These samples were taken from the Large- and SHiGDA groups described above and from 17 patients undergoing colonoscopy for diverse indications. Results: p87 FE were found in 40% of 10 LHiGDA and 0% of 3 SHiGDA patients. The ITS in the 2 representative patients showed unique mutational fields in: KRAS, APC, p53 in the LHiGDA and Jak3, PIK3Ca, p53, APC in the SHiGDA patient, both of whom lacked p87 FE. PCR using the Jak3 primers used in the ITS and subsequent sequencing revealed the same non-synonymous mutation in the serum of a FAP patient after colonic resection and an additional colonic segment of the selected ShiGDA patient but not in his serum. Other Jak3 mutations were found in 1 of 8 LHiGDA, 1 (the selected patient) of 3 SHiGDA, 1 of 7 patients with FAP and 1 of 10 colonoscopy patients with a family history of colorectal cancer with a FE and an untoward outcome. The selected SHiGDA patient subsequently contracted and died of NSCLC adenocarcinoma. The positive LHiGDA and FAP patients had a severely dysplastic anal condyloma and severe pancreatitis, respectively. Overall, the non-synonymous mutation occurring in the non-FAP SHiGDA patient occurred in the absence of p87 FE. Conclusions: In this pilot study we demonstrate the presence of Jak3 mutations likely associated with the lack of p87 expression in patients with high grade dysplastic adenomas and 1 FAP patient. Most of these patients had a clinical course which may have differed from their group members suggesting an altered immune system milieu. If confirmed in SHiGDA and FAP, Jak 3 mutations, associated with the SCID and late onset combined immunodeficiency, may allow for intervention with currently available medications to potentially avert a deleterious clinical outcome.
Citation Format: Martin Tobi, Xiaoping Zhao, Darshana Jhala, Rebecca Rodriguez, Fadi Antaki, Edi Levi, Paula Sochacki, John Lieb, MaryAnn Rambus, Tapan Ganguly, Martin Bluth, Michael J. Lawson. Jak3 mutations in colorectal neoplasia- Preliminary data on a not so silent minority abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3689. doi:10.1158/1538-7445.AM2017-3689
Abstract
Monoclonal antibody Adnab-9 recognizes the p87 antigen, a constituent of putatively activated pulmonary type 2 pneumocytes. p87 shed into sputum and swallowed may be a potential tumor marker ...detectable by the Adnab-9 stool test. We describe prospectively collected stool and colonic effluent ELISA results in a cohort of urban patients enrolled in a colorectal cancer screening study. Methods: Prospectively, 1,132 American patients in Metropolitan Detroit submitted stool in a CRC prospective screening study from 1996 to 2013. Eleven patients free of any significant colorectal neoplasia or other cancer, but who had died from non-cancer-related causes, served as controls. Patients and controls were approximately 50-80 years old and the male to female ratios were similar (30:1 and 10:1). Sixty-seven percent of patients and 45 % of controls were African American. Results: 64 patients of 1,132 contracted lung cancer during the 17 year period of follow-up (5.7%). Of these 64 patients, 34 (52%) had stool or effluent Adnab-9 binding tests on retrospective review. We found that over half the patients ultimately diagnosed with lung cancer had either stool/effluent ELISA or Western blot positivity for Adnab-9 compared to only 1 of 11 controls (OR = 11.3, CI 1.3-97.9, p=0.014). The median time from stool submission to diagnosis of lung cancer was 4.4 (0.7-10.3) years. Lung Cancer Tumor types were adenocarcinoma in 23%; squamous in 33%; other in 20%; large cell cancer in 9%; small cell cancer in 8%; and unknown in 7%. At the last time of follow-up, 70% of patients subsequently diagnosed with lung cancer were deceased. Median survival time was 2.1 years from the time of clinical diagnosis. Conclusion: Adnab-9 sensitivity in this cohort was 52% with specificity of 90.9% for lung cancer and could lead to the diagnosis of lung cancer 4.4 years before clinical diagnosis was made. This supports the notion that accessible, early non-invasive testing combined with effective follow-up studies may lead to earlier diagnosis and treatment of lung cancer, the World's most frequent cancer.
Citation Format: Martin Tobi, Joe Sayre, MaryAnn Rambus, Fadi Antaki, Ann Schwartz, Michael J. Lawson. Non-invasive diagnostic testing for lung cancer in a prospective cohort. abstract. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 882. doi:10.1158/1538-7445.AM2014-882
Abstract
Sporadic pancreatic ductal adenocarcinoma (PDA) is a highly lethal cancer and with no proven screening strategies. Adnab-9, a monoclonal antibody to constituents of human colonic adenomas ...has been shown to be prognostic in IPMN and diagnostic in gastric and colorectal cancer or adenomas (CRN). In this study, we retrospectively and prospectively evaluated the diagnostic potential of Adnab-9 in the stool of two distinct PDA populations. Methods: As proof of principle, 249 Chinese patients submitted stool for screening. Retrospectively, 15 patients had PDA and 80 patients without cancer served as controls. Adnab-9 stool reactivity was determined by ELISA. Prospectively, 1132 American CRN screening program patients submitted stool. Six patients developed PDA over five years of follow-up. 11 Patients free of any significant CRN or other cancer but deceased due to other causes, served as controls. An additional four patients with a known family history of PDA also prospectively submitted stool for Adnab-9. Results: The prospective study results are summarized in the Table. Table: Prospective Study Data for Adnab-9 stool antibody as a biomarker of PDA. The pilot study in Chinese PDA patients found that 12 of 15 patients had stool Adnab-9 ELISA positivity compared to 10 of 80 controls (OR 28, CI 6.7-116.8, P<0.0001). 4 of 6 prospective American study PDA patients had either stool ELISA or Western blot positivity for Adnab-9 compared to 1 of 11 controls (OR=20, CI 1.4-287.6, P<0.03). 2 of 4 patients with family history of PDA had stool positive by Adnab-9 ELISA or Western blot. Conclusions: Our study suggests that Adnab-9 stool tests may be an effective PDA diagnostic marker. Based on previous data in CRC and IPMN, Adnab-9 appears to bind Paneth-like cells, which we and others have shown to be cancer-associated and the likely source of the stool antigen. Future prospective studies are needed to confirm the diagnostic efficacy of Adnab-9 stool testing in PDA.
Citation Format: {Authors}. {Abstract title} abstract. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3699. doi:1538-7445.AM2012-3699
Abstract
Evidence accrues that screening colonoscopy in average-risk patients does not reduce mortality from right-sided neoplasia. A non-invasive test that differentially identifies right-sided ...neoplasia is urgently needed to direct investigative colonoscopy. The metabolic syndrome is associated with excess colorectal neoplasia but many medications that treat the diabetic component of this syndrome may alter carcinogenesis pathways (PPARδ in de novo for thiazolidinediones etc) and may thus confound our ability to design such a test. Methods: We investigated 122 patients, 45 diabetics and 77 non-diabetics at above average risk identified by screening by a risk questionnaire, for colorectal cancer. The patients were enrolled to participate in a prospective colorectal cancer screening study at the Detroit VAMC and intake of metformin, thiazolidinediones, ASA, NSAID, sulfonylureas, statins, insulin, and multivitamins was determined. We ascertained demographics, nutritional parameters (total caloric, protein and vitamin intake, BMI, alcohol intake, fried food, etc). Primary outcomes were colorectal neoplasia (adenomas & cancers) found on colonoscopy. Secondary endpoints were: adenoma-carcinoma biomarker binding in body fluids and mucosal field effects using the Adnab-9 monoclonal antibody. These diagnostic tests were also examined in the cohort of 77 of non-diabetics to ascertain whethera specific test was also applicable to the non-diabetic population at increased risk for CRC. Results: Of 45 diabetic patients with evaluable data, 26.6% had right-sided neoplasia. Of these 50% had positive binding compared to 13.8% without such neoplasia. There was a significant correlation between Adnab-9 effluent binding and right-sided neoplastic lesions (OR 6.45;CI 1.48-26.39;p<0.001). Adnab-9 effluent binding showed a 67% sensitivity and 76% specificity for right sided lesions, True positive=50% and True Negative=86% with a likelihood ratio of 2.75 (CI 1.33-5.67) for a positive test and 0.44 (0.19-0.998) for a negative test. There was no significant differential binding in the non-diabetic population (OR1.74;CI 0.59-5.11). There were also no statistically significant differences between the above medication groups or with other characteristics that included age, diabetes duration, BMI, HbA1c, GFR, and nutritional parameters with respect to neoplasia prevalence. Other adenoma-carcinoma biomarker binding in stool or mucosa was not significantly different between those with or without neoplasia. Conclusions: Adnab-9 Effluent binding may differentially identify most diabetics with right-sided colonic neoplasia independent of medication and other parameters. Prospective studies will be needed to determine the impact of this test in the reduction of mortality from right-sided CRC in this high-risk group.
Citation Format: {Authors}. {Abstract title} abstract. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4519. doi:1538-7445.AM2012-4519
