Single-cell CBD Biomarkers of Inflammation Reduction in People Living With HIV: Complete Statistical Analysis

1. Trial at a Glance

This page covers the registry results for Single-cell CBD Biomarkers of Inflammation Reduction in People Living With HIV (NCT05209867), a na phase 4 study. The primary outcome is Number of Differentially Expressed Genes.

2. Clinical Question

This is a biomarker question, not a question about clinical symptoms, comorbidities, or HIV disease outcomes. The study description proposes examining immune-cell phenotypes and the molecular role of different immune cells. The listed primary endpoint narrows that broad aim to a count of genes meeting a significance criterion for fold change before versus after CBD.

The registry record describes different follow-up timing among the participants: one participant had a 1-month assessment, and two had a 2-month assessment. A fourth participant completed 2 months, but the record says the quality of the data was insufficient. It does not report endpoint values or clarify the full analysis population. Therefore, the clinical question is clear as an intended measurement, but the posted record does not establish what the measured gene-count outcome was.

3. Trial Design

This is a completed Phase 4, single-group intervention study sponsored by the University of Florida. The registry lists enrollment of 4, no masking, and allocation as not applicable. It lists chronic inflammation as the condition and CBD oil as the intervention. Participants were to self-administer CBD daily for 2 months.

The study uses a before-and-after structure: blood measurements at baseline are compared with measurements after CBD exposure. There is no concurrent control group in the record, and no randomized treatment allocation. Consequently, any change over time would be a within-person observation rather than a randomized estimate of the difference between CBD and a control treatment. Such observations can describe biomarker changes, but they cannot by themselves separate a treatment-related change from other explanations, including time-related variation or measurement variability.

The registry lists a start date of 2022-05-01, a primary completion date of 2022-12-12, and a completed status. The brief study description says the investigators planned to study five people, whereas the outcome population description says four individuals completed the assigned intervention. The record does not explain this difference. It states that one individual completed one month, two completed two months, and another completed two months but had data deemed insufficient in quality after protocol and data review.

The registry record does not report a control arm, a masking procedure, or a randomized allocation scheme. It also does not report the protocol details needed to explain the enrollment discrepancy or the consequences of the data-quality review. No statistical analysis is registered.

4. Participant Flow and Accounting

The registry reports how participants moved through each stage of the study. All counts below are exactly as posted; missing counts are not inferred from other stages.

Overall Study

Analysis population for the primary outcome, as defined in the registry: Four individuals completed the assigned intervention. One individual completed one month only. Two individuals completed two months. A fourth individual completed two months but, after protocol and data review, the quality of the provided data was deemed not sufficient (single-cell RNA was not sequenced, gene expression not measured for that individual).

5. Baseline Characteristics

The table reproduces the baseline characteristics posted in the registry, which describe the participants analysed at baseline and help show whether the randomized groups were comparable.

Population: Two individuals completed the assigned intervention. A third individual completed the first month of the assigned intervention. A fourth individual completed the assigned intervention but, after review, the quality of the provided data was deemed not sufficient (single-cell RNA was not sequenced, gene expression not measured).

6. Primary Endpoint

The registry lists the primary outcome as Number of Differentially Expressed Genes, measured over Two time points considered: baseline, and after 1 month (1 individual) or 2 months (2 individuals).

The primary endpoint is the number of differentially expressed genes: a count of genes related to inflammation that had a significant fold change in peripheral blood mononuclear cells before versus after CBD treatment. The record says these measurements were based on single-cell RNA sequencing. It describes baseline and follow-up assessments, with one participant assessed after 1 month and two after 2 months.

A gene-count endpoint can condense a large molecular dataset into a measure of how many genes cross a specified threshold for change. It is intended to capture a pattern of inflammation-related molecular activity, rather than a direct clinical outcome. The count alone does not indicate which genes changed, the direction or size of each change, or whether a measured change corresponds to a meaningful change in health.

The record does not specify the threshold defining a significant fold change, the gene-level statistical procedure, or the endpoint values. It also does not report means or standard deviations for the count. Although the outcome is described as a mean with standard deviation, the posted results do not provide the corresponding numerical summary. The biological aim is stated, but the reported endpoint information is not sufficient to assess the magnitude or precision of an observed change.

7. Why No Re-analysis Is Shown

8. Statistical Interpretation

9. Statistical Methods Explained

The record describes a continuous outcome summarized as a mean and standard deviation, but it does not report an actual mean or standard deviation for the primary endpoint. The endpoint itself is a count of inflammation-related genes meeting a fold-change significance criterion. The measurement process uses single-cell RNA sequencing of peripheral blood mononuclear cells, with baseline and post-treatment observations.

What does a before-and-after comparison estimate?

In general, a before-and-after design compares each participant's measurement at follow-up with that participant's own baseline. Pairing can account for stable differences between people because each person serves as their own reference. However, the approach does not by itself distinguish a treatment-associated change from other changes occurring over time. A concurrent control group can help address some alternative explanations; none is listed in this registry record.

Assessment timing is not uniform in the outcome description. One participant had a 1-month assessment, while two had 2-month assessments. These are not identical follow-up contrasts. A mean that combines different follow-up times would need a clear definition and analysis rationale, but the registry record does not report how the observations were combined or analyzed. It also does not report whether the endpoint was calculated separately for each participant before any group summary.

What do the mean and standard deviation convey?

In general, a mean describes the average value of a continuous measurement across the analyzed observations. A standard deviation describes how spread out those values are around the mean. These are descriptive summaries; neither alone establishes whether a change is statistically distinguishable from chance or whether it is clinically important. The registry identifies these summary measures for the endpoint but provides no corresponding values.

A confidence interval, in general, expresses the uncertainty around an estimated quantity under a specified statistical model. Its width depends on the data and analysis assumptions. No confidence interval is reported for this endpoint, so the precision of an estimated average cannot be assessed from the registry record. A p-value, in general, measures how compatible data are with a specified null hypothesis under a statistical procedure; it does not express the size or clinical importance of an effect. No endpoint p-value or hypothesis test is reported.

Why is the gene-level procedure important?

In general, testing many genes creates a multiplicity problem: when many hypotheses are assessed, some may meet a nominal significance threshold by chance even if there is no true change. Statistical approaches can control different error rates or otherwise account for the number and structure of tests. The endpoint description says genes were counted when they had a significant fold change, but it does not report the gene-level threshold, correction method, model, or other decision rules. The count therefore cannot be interpreted as a reproducible tally of statistically supported changes without those details.

Single-cell data also have a hierarchical structure: cells are measured within people, and cells from the same person are not generally independent observations. In general, treating all cells as independent can overstate the amount of information and yield uncertainty estimates that are too narrow. Analysis at the participant level or a method that accounts for within-person dependence may be appropriate, depending on the scientific question and design. The registry does not state how cell-level data were aggregated or how dependence was addressed.

What can the planning assumptions tell readers?

The registry includes general assumptions for planning illustrations: a two-sided significance level of 0.05, equal allocation between two arms, and a normal approximation. Those assumptions describe a planning calculation, not the single-group design listed for this study. The illustrations are based on a fraction of an observed effect and are not observed power. The registry record does not report the trial's own sample-size justification or explain whether these assumptions were part of the study's design.

Sample-size illustrations are conditional on their assumptions and should not be read as evidence that the study had a particular probability of detecting an effect. They cannot resolve uncertainty when the observed endpoint estimate and its variability are not reported. The registry also does not document a statistical analysis plan, interim analyses, or a prespecified method for addressing incomplete or unusable measurements.

These general statistical considerations help identify what would be needed to interpret the endpoint: a defined gene-level criterion, a transparent data-processing and analysis approach, participant-level summaries, an explanation of follow-up timing and data exclusions, and uncertainty estimates. The registry record does not report those details or the numerical outcome itself.

10. Confidence Intervals for Each Arm

No two-arm comparison is shown for this trial, but each arm's mean can be given with a t-based 95% confidence interval computed from the posted mean, spread and number of participants.

R code and output

arm <- c("CBD Intervention")
n   <- c(3)
m   <- c(8.3)
sp  <- c(7)
sd  <- sp      # posted standard deviations
for (i in seq_along(n)) {
  se <- sd[i] / sqrt(n[i]); h <- qt(0.975, n[i] - 1) * se
  cat(sprintf("CARM|%s|%d|%.3f|%.3f|%.3f|%.3f\n", arm[i], n[i], m[i], sd[i], m[i] - h, m[i] + h))
}

CARM  CBD Intervention  3  8.300  7.000  -9.089  25.689

11. Multiplicity, Missing Data and Analysis Populations

The registry lists one primary outcome measure and no registered analyses. That does not establish whether investigators conducted no analyses; it means that analysis details are not reported in the registry record. The endpoint is a count derived from gene-level significance decisions, but the number of genes tested, the significance criterion, and any multiplicity adjustment are not reported. In general, testing many genes raises the chance of false-positive findings unless the analysis accounts for multiple comparisons. The record does not say how that issue was handled.

Missing and unusable data also affect interpretation. The outcome description says one participant completed one month, two completed two months, and a fourth completed two months but had data deemed insufficient in quality. It does not report the exact analysis population, how the quality decision was made, or whether any measurements were excluded beyond that statement. The study brief says five people were planned, while the outcome description refers to four intervention completers; the record does not reconcile the difference.

The registry does not report an imputation method, a missing-data assumption, or a sensitivity analysis. It does not state whether participants with different follow-up times were analyzed separately or together. These omissions make it difficult to assess how incomplete follow-up, data quality, and analysis choices might affect the reported endpoint. No analysis population such as an intention-to-treat or per-protocol set is defined in the record.

12. Safety Results

The registry reports serious adverse events as affected participants divided by participants at risk.

Deaths

Other (non-serious) adverse events

Participants with at least one non-serious event at or above the registry's 0% reporting threshold.

Most frequent non-serious adverse event terms

The adverse-event record lists four participants at risk for serious adverse events and zero serious adverse events. It lists four at risk for other adverse events and one other adverse event. The death count is zero, and no serious adverse-event terms are listed. These are counts, not estimates of comparative risk.

Because this is a single-group study with no listed control arm, the counts cannot compare CBD with another treatment or with no treatment. The record does not give details about the nature, timing, severity, or relatedness of the other adverse event. It also does not describe the criteria or collection procedures used for adverse-event reporting.

A zero count in the registry means no event was recorded in that category in the posted results; it does not establish that the risk is absent. The small enrolled group and the limited information about event ascertainment constrain what can be concluded about safety. These figures do not address the primary gene-expression endpoint and should not be used to infer whether CBD changes inflammation-related biomarkers.

13. What the Result Does, and Does Not, Mean

The study asks whether inflammation-related gene expression in peripheral blood mononuclear cells differs between baseline and after daily CBD use, using single-cell RNA sequencing. Its primary endpoint is a count of genes with a significant fold change. This is a molecular biomarker outcome, not a direct measure of symptoms, comorbidities, or other clinical outcomes.

The registry record does not report the endpoint's numerical results, so it does not establish the direction or size of any change, the variability across participants, or the precision of an estimate. It also does not specify the gene-level significance criterion or the approach to multiple testing. Without these elements, readers cannot evaluate how robust or reproducible any gene-count result would be.

Even if a before-and-after biomarker change had been reported, the single-group design would not establish that CBD caused it. There is no randomized control group to distinguish treatment-associated changes from other influences or natural variation over time. The differing follow-up times and the data-quality issue further limit interpretation of the outcome population.

The record therefore documents a research question and an intended molecular measurement, but it does not provide sufficient posted results to answer that question quantitatively. It cannot support a conclusion that CBD does or does not reduce inflammation, nor can it establish clinical benefit or harm. The adverse-event counts are limited descriptive information and do not substitute for a comparative safety analysis.

14. Why This Trial Matters Statistically

This study is statistically notable because it connects a single-group clinical intervention with single-cell molecular measurements. Such data can reveal patterns across immune cells, but the analysis must respect the fact that cells are nested within participants. The number of cells measured is not the same as the number of independent people contributing evidence about a treatment effect. The registry does not report how this dependence was handled.

The endpoint also turns many gene-level measurements into a count of genes passing a significance criterion. That creates an interpretive dependency on the threshold, test procedure, and multiplicity strategy. Without those details, the count is difficult to compare across analyses or to reproduce. Reporting the gene-level method and participant-level summaries would clarify what the endpoint represents.

The design makes the distinction between biomarker observation and causal inference especially important. Baseline measurements offer a within-person reference, but no randomized control group is listed. Follow-up timing varies, and one participant's data were considered insufficient in quality. These features matter for the definition of the analysis population and for the uncertainty attached to any summary.

The registry record does not post the numerical primary outcome or a statistical analysis. As a result, the study is useful for identifying methodological questions about small single-group biomarker studies, but its registered information does not allow readers to assess the observed gene-expression signal. Clear reporting of outcome estimates, uncertainty, analysis choices, and data exclusions would be necessary for a fuller statistical interpretation.

15. Limitations

• The posted summary statistics are the only data these results rest on; patient-level data, full protocols and statistical analysis plans are not available.
• Recalculated results are simple, unadjusted calculations from posted summaries. They do not replace the sponsor's prespecified analysis.

Several limitations are specific to the information recorded for this study. First, the primary outcome results are not reported. There is no numerical gene count, mean, standard deviation, confidence interval, or p-value in the registry record. The direction and magnitude of any before-and-after change are therefore unknown from the posted information.

Second, the study has no listed control group or randomized allocation. A before-and-after pattern cannot isolate the effect of CBD from changes that could occur over time for other reasons. The lack of masking is also recorded, while the registry does not describe any steps intended to limit measurement or analytic bias.

Third, assessment timing and data quality are not uniform in the outcome description. One person had a 1-month assessment, two had 2-month assessments, and a fourth completed 2 months but had data judged insufficient in quality. The registry does not provide the analysis population, explain the quality review, or say how the resulting missing or unusable data were handled. The brief study description's plan for five people is not reconciled with the outcome description of four completers.

Finally, key analytical details are absent. The registry does not state the gene-level significance threshold, multiplicity adjustment, model, cell-level aggregation method, or method for accounting for measurements clustered within participants. It also does not report a statistical analysis plan. These gaps prevent evaluation of both the endpoint's reproducibility and the precision of any claimed result.

16. Related Tutorials

17. Related Statistical Calculators

18. Sources

• ClinicalTrials.gov: Single-cell CBD Biomarkers of Inflammation Reduction in People Living With HIV — NCT05209867.
• Registry record last updated 2023-12-28; status last verified 2023-12.
• Results first posted 2023-12-28.
• Data retrieved from ClinicalTrials.gov on 2026-11-09.
• Group sizes, means, event counts and sponsor-reported p-values are reproduced as posted. Effect estimates, confidence intervals and p-values not attributed to the sponsor are independently calculated from the posted summary statistics.

19. Record Summary

The registry record describes a completed Phase 4, single-group study of daily CBD oil in people living with HIV and chronic inflammation. It lists enrollment of 4, no masking, and no randomized allocation. The primary endpoint is the count of inflammation-related genes with significant fold change in peripheral blood mononuclear cells before versus after treatment, measured using single-cell RNA sequencing.

The outcome description records one participant with a 1-month assessment and two with 2-month assessments. A fourth participant completed 2 months, but the data were deemed insufficient in quality. The study brief describes a plan to study five people, and the registry does not explain the difference from the outcome population description.

The record does not report numerical primary outcome results, a statistical analysis, or details of the gene-level testing and missing-data handling. It lists zero serious adverse events, one other adverse event, and zero deaths. Those safety counts are descriptive and do not provide a comparison with another group.