Collaborating to Compete: Blood Profiling Atlas in Cancer (BloodPAC) Consortium.

The cancer community understands the value of blood profiling measurements in assessing and monitoring cancer. We describe an effort among academic, government, biotechnology, diagnostic, and pharmaceutical companies called the Blood Profiling Atlas in Cancer (BloodPAC) Project. BloodPAC will aggregate, make freely available, and harmonize for further analyses, raw datasets, relevant associated clinical data (e.g., clinical diagnosis, treatment history, and outcomes), and sample preparation and handling protocols to accelerate the development of blood profiling assays.

Current Status of Companion and Complementary Diagnostics: Strategic Considerations for Development and Launch.

US Food and Drug Administration (FDA)-approved diagnostic assays play an increasingly common role in managing patients to prolong lifespan while also enhancing quality of life. Diagnostic assays can be essential for the safe and effective use of therapeutics (companion diagnostic), or may inform on improving the benefit/risk ratio without restricting drug access (complementary diagnostic). This tutorial reviews strategic considerations for drug and assay development resulting in FDA-approved companion or complementary diagnostic status.

Adaptive Design for a Confirmatory Basket Trial in Multiple Tumor Types Based on a Putative Predictive Biomarker.

Increasingly, tumors are defined on a molecular basis rather than only on histology, and targeted agents, which address these molecular subtypes, are being approved. This profusion of molecular subtypes creates "rare" diseases as subsets of common cancers, leading to difficulties in enrolling sufficiently large cohorts for confirmatory trials. However, if the molecular subtype is shared across various histologies, these may be pooled into a basket trial. To date, basket trials have been primarily for exploratory early development. In this perspective, we consider qualitative designs for confirmatory basket trials. These confirmatory basket designs will provide patients in niche indications with enhanced access to novel therapies, facilitate development and full approval for niche indications, allow accelerated approval for indications within a basket based on a surrogate endpoint, reduce development cost by combining trials, and enhance the ability of regulatory authorities to evaluate risk and benefit in niche indications.

Using 16O35Cl to correct for chloride interference improves accuracy of urine arsenic determinations by inductively coupled plasma mass spectrometry.

We have observed inaccurate urine arsenic values with the method of isobaric fractionation, which was designed to correct for the 40Ar35Cl interference with 75As quantitation by inductively coupled plasma mass spectrometry. Isobaric fractionation, which is based on ion intensities at m/z 77 and 82, consistently underestimates the 40Ar35Cl interference and overestimates urine arsenic. We present an improved method for identifying the argon-chloride interference. We observed that signal intensities for the species 16O35Cl and 40Ar35Cl are proportional (I75 = 0.0295 x I51 - 14.7, r2 = 0.998; where Ix is the normalized ion intensity at m/z X) in water and urine, over a broad range of chloride concentrations (0-800 mmol/L). The proportionality constant is remarkably stable within a run (mean and SD, 0.0295 +/- 0.0023, based on 10 replicates of five chloride calibrators, 0, 100, 200, 400, and 800 mmol/L). Increased sensitivity (50-fold) for detecting the 40Ar35Cl interference provides improved accuracy for urine arsenic quantitation as demonstrated by a split-sample comparison with graphite-furnace atomic absorption spectrophotometry.

A simple ICP-MS procedure for the determination of total mercury in whole blood and urine.

A simple and sensitive procedure for total mercury in whole blood and urine using inductively coupled plasma-mass spectrometry (ICP-MS) is described. Specimens are prepared by precipitation-extraction with 50% v/v hydrochloric acid containing EDTA and cysteine, centrifuged, and filtered through fritended screening column; the filtrates are directly analyzed by ICP-MS. The method is linear between 2 and 200 micrograms/L in the specimen with an absolute sensitivity of 0.2 microgram/L in the final supernatant. The assay variability at various concentrations (microgram/L) of mercury are as follows: intra-assay whole blood (n = 20)-4.6 +/- 0.6 (c.v. 12.3%), 18.3 +/- 1.1 (c.v. 6.1%), 56.4 +/- 2.8 (c.v. 5.0%); inter-assay whole blood (n = 15)-5.7 +/- 1.0 (c.v. 16.8%), 19.7 +/- 2.7 (c.v. 13.5%), and 50.1 +/- 6.9 (c.v. 13.7%); urine (n = 20)-9.3 +/- 1.2 (c.v. 12.9%), 29.6 +/- 2.2 (c.v. 7.4%). Recovery of organic and inorganic mercury from blood samples ranges from 91.6% to 110.2%. The method is suitable for analysis of total mercury, both organic and inorganic, in whole blood and urine.

O6-methylguanine (O6-MeG) and cytotoxicity: reversion analysis involving an N-methyl-N'-nitro-N-nitrosoguanidine-sensitive, O6-MeG-DNA methyltransferase-deficient HeLa cell mutant.

In a previous communication, we proposed that N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced cytotoxicity in an O6-methylguanine (O6-MeG)-DNA methyltransferase-deficient (MT-) HeLa cell mutant was mainly the consequence of DNA strand breaks resulting from the failure to remove O6-MeG lesions. If MNNG-induced cytotoxicity, DNA strand breaks and O6-MeG lesions are related, there should be a corresponding relationship of these properties in MNNG-resistant clones derived from the MT- strain. A study of such revertants indicated that they were a heterogeneous group with increased repair of DNA strand breaks and O6-MeG lesions and increased resistance to the cytotoxic effects of MNNG. These observations support the hypothesis relating O6-MeG, DNA strand breaks and cytotoxicity. The relationship of these 'revertants' to the MT- and wild-type strains is discussed.

O6-methylguanine-DNA methyltransferase-defective human cell mutant: O6-methylguanine, DNA strand breaks and cytotoxicity.

We have isolated an isogenic O6-methylguanine (O6-MeG)-DNA methyltransferase-defective mutant from a HeLa cell line. This mutant exhibits excess DNA strand breaks and considerable cytotoxicity after N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) treatment. The increased frequency of strand breaks after MNNG treatment was not abolished by DNA synthesis inhibitors. We propose that the presence of unrepaired O6-MeG lesions leads to excess strand breaks and these, in turn, are mainly responsible for the cytotoxicity.

Fat and cancer.

Experimental studies and epidemiologic data both indicate that high-fat diets increase the risk of cancer at sites such as breast, colon, and pancreas. Dietary polyunsaturated vegetable oils promote tumorigenesis in animals whereas saturated fats and polyunsaturated fish oils either have little effect or are inhibitory. A blend of fats typical of the American diet enhanced mammary tumorigenesis effectively when fed as 40% of calories. After 9 to 10 weeks on this diet, reducing the fat to 10% of calories inhibited subsequent tumor development. Although dietary fat appears to act as a promoter, the exact mechanisms are still unknown. Cancer mortality in humans correlates better with total dietary fat than with degree of unsaturation, perhaps because most diets contain more than the amount of polyunsaturated fatty acids required for maximum effect on tumorigenesis in animals. A reduction in total dietary fat is recommended to reduce the current high cancer mortality.

Hepatic drug metabolising enzymes in undernourished men.

The influence of the nutritional status of the individual on the hepatic drug metabolising enzymes in human wedge-biopsy liver samples was investigated. The aryl hydrocarbon hydroxylase (AHH) was found to be elevated although there was no change in the cytochrome P-450 levels in the undernourished groups. Conjugating enzymes viz. UDP glucuronyl transferase (UDPGT) and glutathione-S-transferase (GST) were not found to be different. The increase in AHH activity as compared to UDPGT and GST activities suggests that the activation processes may possibly be increased in the undernourished segment of the population.

Dietary influences on the kinetics of antipyrine and aminopyrine in human subjects.

The possible dietary influences on in vivo antipyrine and aminopyrine kinetics with reference to energy (1500, 1800, 3000 kcal) and protein (5, 10, 15, 20% protein energy-PE) intake were studied in a carefully controlled metabolic experiment in young healthy adult male volunteers aged between 25-34 years. Antipyrine and aminopyrine were used to evaluate drug metabolism. On 1500 kcal intake with 10% PE, the metabolism of both aminopyrine and antipyrine were significantly reduced whereas on 1800 kcal with 10% PE intake, only aminopyrine metabolism decreased significantly as compared to 3000 kcal with 10% PE. Antipyrine clearance on 1800 kcal with 10% PE however had not decreased to the same extent as on 1500 kcal with 10% PE. The results indicate that on low calorie intake with 10% PE, the drug metabolism is decreased. When the protein intake on 1500 kcal was doubled (20% PE) there was a significant stimulation of both aminopyrine and antipyrine metabolism. Aminopyrine and antipyrine clearances on 3000 kcal with 5% PE were significantly reduced as compared to 3000 kcal with 10% and 15% PE indicating that unlike proteins, carbohydrate and/or fat calories per se do not significantly stimulate drug metabolism. When the protein energy in the diet was increased from 5% to 10% or 15% at 3000 kcal, there was a stimulation of both antipyrine and aminopyrine metabolism. Significant differences between 10% and 15% of protein energy were not observed when the energy was adequate (3000 kcal). Therefore it is necessary to consider both proteins and energy as important variables affecting drug clearances from plasma in malnourished conditions.

Reversal of the promotional effect of high-fat diet on mammary tumorigenesis by subsequent lowering of dietary fat.

Female Sprague-Dawley rats were given 7,12-dimethylbenz(a)anthracene at 50 days of age to induce mammary tumors, and beginning one week later were fed a high-fat, semipurified diet containing 20% sunflowerseed oil to promote tumor development. After another 7 weeks, when one third of the rats had palpable mammary tumors, the rats were randomly assigned to five groups of 31 animals each, with the same number of tumor-bearers in each group. One group was continued on the high-fat diet, another was given a fat-free diet, and the three remaining groups were fed diets containing 10% lard, butter, or coconut oil, respectively. During the next 29 weeks, rats fed the diets containing 0% or 10% fat developed significantly fewer tumors than those continued on the 20% fat diet. The diets containing 10% fat suppressed tumorigenesis at least as effectively as the fat-free diet. Rats fed the 10% butter and 10% lard diets had growth rates comparable to those fed the 20% sunflowerseed-oil diet throughout, and evidence of essential fatty acid deficiency was seen only in rats on the fat-free diet. These results provide additional evidence that high-fat diets promote development of mammary cancer and suggest that reducing the level of dietary fat might help to prevent the development and recurrence of breast cancer in humans.

Metabolism of drugs and carcinogens in man: antipyrine elimination as an indicator.

The suitability of the most commonly used "prototype" drug, viz, antipyrine, in predicting drug and carcinogen metabolism was evaluated, by studying in vivo antipyrine elimination rate (Ke) and in vitro metabolism of drugs and carcinogens in liver preparations in the same individuals. Our subjects were 20 adult males undergoing abdominal surgery for gastrojejunostomy, although antipyrine Ke could be studied in only 16 subjects. Correlations of the various in vito--in vivo parameters were positive between the parameter pairs: in vivo antipyrine Ke--in vitro benzopyrene hydroxylase; benzopyrene hydroxylase--aniline hydroxylase; and benzopyrene hydroxylase--gamma-glutamyl transferase. Aminopyrine demethylase did not correlate with any of the parameters studied. The degree of correlation between antipyrine Ke and benzopyrene hydroxylase was statistically significant but was not satisfactory for predictive purposes. Our study indicates some of the problems and limitations of in vivo--in vitro comparisons and confirms earlier doubts on the usefulness of antipyrine as a "prototype" drug for predicting drug and carcinogen metabolism in man.

Functional significance of low serum vitamin B12 levels in pregnancy.

The physiological significance of low levels of vitamin B12 in serum in late pregnancy was assessed by a functional test viz. urinary excretion of methylmalonic acid (MMA) after a valine load. This was normal in all but one subject despite low levels of vitamin B12 in serum in many of these subjects. Assay of serum vitamin B12 levels in pregnant women and control subjects by two different techniques viz. radio isotope dilution and microbiological methods gave essentially similar results for both groups, although the absolute values in both the groups were over-estimated by the former technique. The data presented indicate that there is no evidence of functional inadequacy in pregnant women with low levels of vitamin B12 in serum.

Tropical ataxic myelopathy.

A syndrome of ataxic myelopathy with optic atrophy and perceptive deafness has been described. This is essentially similar to the Nigerian ataxic myelopathy which has been ascribed to chronic cyanide intoxication through dietary means. The results of biochemical investigations and bone marrow examination indicate that neither vitamin B12 deficiency nor cyanide intoxication are likely factors in the detiology of our cases. It is possible that the disease may be due to an unrecognized toxic for factor which may or may not be of dietary origin.