Assessment of Type I Error Rates and Power of Common Analysis Methods in Murine Obesity-Related Study: ‘Plasmode-Based’ Simulation (P13-011-19)

Abstract

ObjectivesRigor, reproducibility and transparency (RRT) awareness has expanded over the last decade. Although RRT can be improved from various aspects, we focused on type I error rates and power of commonly used statistical analyses testing mean differences of two groups, using small (n ≤ 5) to moderate sample sizes. MethodsWe compared data from five distinct, homozygous, monogenic, murine models of obesity with non-mutant controls of both sexes. Baseline weight (7–11 weeks old) was the outcome. To examine whether type I error rate could be affected by choice of statistical tests, we adjusted the empirical distributions of weights to ensure the null hypothesis (i.e., no mean difference) in two ways: Case 1) center both weight distributions on the same mean weight; Case 2) combine data from control and mutant groups into one distribution. From these cases, 3 to 20 mice were resampled to create a ‘plasmode’ dataset. We performed five common tests (Student’s t-test, Welch’s t-test, Wilcoxon test, permutation test and bootstrap test) on the plasmodes and computed type I error rates. Power was assessed using plasmodes, where the distribution of the control group was shifted by adding a constant value as in Case 1, but to realize nominal effect sizes. ResultsType I error rates were unreasonably higher than the nominal significance level (type I error rate inflation) for Student’s t-test, Welch’s t-test and permutation especially when sample size was small for Case 1, whereas inflation was observed only for permutation for Case 2. Deflation was noted for bootstrap with small sample. Increasing sample size mitigated inflation and deflation, except for Wilcoxon in Case 1 because heterogeneity of weight distributions between groups violated assumptions for the purposes of testing mean differences. For power, a departure from the reference value was observed with small samples. Compared with the other tests, bootstrap was underpowered with small samples as a tradeoff for maintaining type I error rates. ConclusionsWith small samples (n ≤ 5), bootstrap avoided type I error rate inflation, but often at the cost of lower power. To avoid type I error rate inflation for other tests, sample size should be increased. Wilcoxon should be avoided because of heterogeneity of weight distributions between mutant and control mice. Funding SourcesThis study was supported in part by NIH and Japan Society for Promotion of Science (JSPS) KAKENHI grant.