Phase I, Open Label Dose Escalation Study to Evaluate Safety of iHIVARNA-01 in Chronically HIV-infected Patients: Complete Statistical Analysis

1. Trial at a Glance

This page covers the registry results for Phase I, Open Label Dose Escalation Study to Evaluate Safety of iHIVARNA-01 in Chronically HIV-infected Patients (NCT02413645), a non-randomized phase 1 study. The primary outcome is Dose Limiting Toxicity (DLT).

2. Clinical Question

The study also listed immunogenicity outcomes based on HIV-1-specific responses to IN and OUT peptide pools, measured by ELISPOT at baseline and weeks 4, 6, 8 and 24, as well as an outcome concerning the effect on the reservoir. The registry record does not provide results for those outcomes. Accordingly, the posted information supports a description of the study question and the safety counts, but not an assessment of immune response or reservoir effects.

3. Trial Design

This was a Phase I, open-label, non-randomized, sequential dose-escalation study. Its title and summary describe safety evaluation and establishing a recommended dose for iHIVARNA-01. The registry lists enrollment of 21 and a completed status. The recorded study dates are a start in 2015-06 and primary completion in 2016-06.

The five listed cohorts cover 100 μg TriMix mRNA, 300 μg TriMix mRNA, 600μg mRNA comprising 300 μg HIV mRNA and 300 μg TriMix mRNA, 900μg mRNA comprising 600 μg HIV mRNA and 300 μg TriMix mRNA, and 1200μg mRNA comprising 900 μg HIV mRNA and 300 μg TriMix mRNA. The descriptions specify 3 patients in each of the first four cohorts, with 6 in the last cohort under the stated condition. They describe escalation decisions in relation to DLTs, including consultation and stopping criteria, and a dose reduction rule for the 900μg cohort. These are protocol descriptions, not evidence that any particular escalation decision occurred.

There is an inconsistency in the registry’s description of the 300 μg TriMix cohort: its arm title identifies 300 μg, while its description says patients receive 100 μg of TriMix mRNA. The record does not resolve this discrepancy. No masking was used. Sequential dose cohorts can inform tolerability at the regimens studied, but their outcomes are not randomized comparisons. The record does not report a sample-size justification, interim analyses, or a fuller statistical analysis plan.

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

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.

6. Primary Endpoint

The registry lists the primary outcome as Dose Limiting Toxicity (DLT), measured over week 24.

The primary endpoint was dose-limiting toxicity, measured as the number of participants with a qualifying event by week 24. The definition includes Grade 3 or above local adverse events, Grade 3 or above systemic adverse events, other Grade 3 or above clinical or laboratory adverse events confirmed by examination or repeat testing, and any vaccination-attributable event leading to discontinuation of the immunization regimen.

This kind of safety endpoint is used in early-phase dose evaluation because it focuses attention on events considered serious enough by the protocol to affect dose escalation or continued immunization. It is a binary participant-level outcome: each participant either meets the endpoint definition or does not. A count is meaningful only alongside the number assessed and a clear, consistently applied definition. The registry record supplies the definition and timeframe, but does not report DLT counts, the number assessed for DLT in each cohort, or how incomplete follow-up was handled.

The adverse-event summaries separately report serious adverse events and other adverse events. Those summaries do not establish which events met the protocol’s DLT definition. In particular, the posted serious and other adverse-event counts cannot substitute for the primary endpoint results. Without a DLT result by cohort, the primary safety question remains unanswered in the registry record.

7. Why No Re-analysis Is Shown

8. Statistical Interpretation

9. Statistical Methods Explained

The posted information is best read as a descriptive account of a small, sequential early-phase study. No registry analyses are listed. There are no reported tests, model estimates, confidence intervals, or adjusted comparisons for the primary endpoint. The registry record does not report a fuller statistical analysis plan.

What does a binary endpoint mean?

In general, a binary endpoint classifies each participant according to whether a defined event occurred. For DLT, that classification depends on the protocol definition and the assessment period. A participant with an adverse event is not automatically a participant with a DLT: the event must satisfy the specified severity, confirmation, attribution, or discontinuation criteria. The registry provides the DLT definition and week 24 timeframe, but does not post the cohort-level DLT classifications.

For an event count to be interpreted, it should be read with its denominator: the number of participants assessed under the same endpoint definition. The safety summary does give numbers at risk and numbers affected for serious and other adverse events. Those denominators relate to those categories in the posted summary; they should not be assumed to be the analysis denominators for DLT unless the registry explicitly says so. It does not.

Why does the sequential design matter?

The registry describes dose escalation in cohorts and gives rules involving DLT counts, including when to consult a data monitoring board, stop, or reduce the dose. In general, a sequential design uses observations in earlier cohorts to guide later enrollment or dose decisions. That makes the order of cohorts part of the design. It is not equivalent to assigning participants at random across dose groups, and the later cohorts are not automatically valid controls for the earlier ones.

General statistical principle: randomization helps balance measured and unmeasured participant characteristics across comparison groups, while masking can reduce some assessment biases. Neither randomization nor masking was used. This does not invalidate descriptive safety reporting, but it limits causal interpretation of differences between cohorts. The registry does not report whether other features of care or assessment were held constant across cohorts.

What uncertainty can be assessed?

In general, a confidence interval describes a range of values compatible with the observed data under a statistical method and its assumptions. For a binary outcome with few participants, that range can be wide, so a count alone gives an incomplete impression of precision. No confidence intervals for the posted adverse-event counts are reported in the registry record. No p-values or statistical comparisons are reported either. Their absence should not be replaced with a claim that the groups are equivalent or that a difference is statistically meaningful.

The record includes a planning illustration based on a two-sided significance level of 0.05, equal allocation between two arms, a normal approximation, and an effect specified as a fraction of the observed effect. It describes time-to-event planning quantities as total events and other planning quantities as participants per arm. These are hypothetical planning assumptions, not the trial’s own design or observed power. The trial itself used sequential, non-randomized cohorts, not equal allocation between two arms. A sample-size illustration cannot remedy the lack of randomized comparisons or supply missing endpoint results.

How should the adverse-event summaries be separated from DLT?

Serious adverse events and other adverse events are separate reported categories. The primary endpoint uses a distinct protocol definition that includes specific severity thresholds and other criteria. General statistical principle: outcomes with different definitions should not be treated as interchangeable, even when they concern the same participants. The registry does not provide event-level details that would allow the serious or other adverse-event summaries to be reclassified under the DLT definition.

The study also lists several secondary outcome areas and repeated assessment times for the ELISPOT outcomes. Repeated measures can be analyzed in different ways, and the method chosen can affect interpretation. The registry record does not state how such measurements were analyzed, how within-participant correlation was handled, or how missing observations were addressed. Those methods cannot be inferred from the outcome descriptions alone.

10. Exact Estimates for Each Arm

No two-arm comparison is shown for this trial, but each arm's result can still be summarized with exact interval estimates computed from the posted counts. The Clopper-Pearson interval is exact and conservative; the Wilson interval is a common closer-to-nominal alternative; the Bayesian credible interval uses a uniform Beta(1, 1) prior. For small groups the three can differ noticeably.

R code and output

arm    <- c("100 μg TriMix mRNA (TriMix_100)", "300 μg TriMix mRNA (TriMix_300)", "600μg mRNA (300 μg HIV mRNA+300 μg TriMix mRNA)", "900μg mRNA (600 μg HIV mRNA+300 μg TriMix mRNA)", "1200μg mRNA(900 μg HIV mRNA+300 μg TriMix mRNA)")
n      <- c(3, 3, 3, 3, 6)
events <- c(0, 0, 0, 0, 0)
for (i in seq_along(n)) {
  ci <- binom.test(events[i], n[i])$conf.int          # Clopper-Pearson exact interval
  cr <- qbeta(c(0.025, 0.975), 1 + events[i], 1 + n[i] - events[i])   # Bayesian, uniform Beta(1,1) prior
  wi <- (function(x, n) { p <- x / n; z <- qnorm(0.975); d <- 1 + z^2 / n
    c((p + z^2 / (2 * n) - z * sqrt(p * (1 - p) / n + z^2 / (4 * n^2))) / d, (p + z^2 / (2 * n) + z * sqrt(p * (1 - p) / n + z^2 / (4 * n^2))) / d) })(events[i], n[i])
  cat(sprintf("ARM|%s|%d|%d|%.1f|%.1f|%.1f|%.1f|%.1f|%.1f|%.1f\n", arm[i], n[i], events[i], 100 * events[i] / n[i],
              100 * ci[1], 100 * ci[2], 100 * wi[1], 100 * wi[2], 100 * cr[1], 100 * cr[2]))
}

ARM  100 μg TriMix mRNA (TriMix_100)  3  0  0.0  0.0  70.8  0.0  56.1  0.6  60.2
ARM  300 μg TriMix mRNA (TriMix_300)  3  0  0.0  0.0  70.8  0.0  56.1  0.6  60.2
ARM  600μg mRNA (300 μg HIV mRNA+300 μg TriMix mRNA)  3  0  0.0  0.0  70.8  0.0  56.1  0.6  60.2
ARM  900μg mRNA (600 μg HIV mRNA+300 μg TriMix mRNA)  3  0  0.0  0.0  70.8  0.0  56.1  0.6  60.2
ARM  1200μg mRNA(900 μg HIV mRNA+300 μg TriMix mRNA)  6  0  0.0  0.0  45.9  0.0  39.0  0.4  41.0

11. Multiplicity, Missing Data and Analysis Populations

The registry lists a primary safety endpoint and secondary outcomes concerning IN and OUT peptide-pool immunogenicity and the reservoir. The ELISPOT outcomes are scheduled at baseline and weeks 4, 6, 8 and 24. Considering several outcomes and assessment times creates opportunities for multiple statistical comparisons. In general, the more comparisons made, the greater the chance that an apparently notable result may arise by chance if each is interpreted independently. The record does not report a multiplicity strategy, such as a prespecified hierarchy or adjustment procedure.

The registry reports 4 outcome measures and 0 analyses. No analysis populations are described, and the record does not report whether analyses were intended to include all enrolled participants, participants who received an immunization, or participants with complete follow-up. It also does not report a method for handling missing outcome values or incomplete assessments. The absence of these details means that the population and data contributing to any result cannot be fully assessed from the registry record.

For the primary endpoint, the record identifies week 24 as the timeframe but does not report the number assessed at that time or the DLT count. For the repeated immunogenicity outcomes, the registry lists assessment times but does not report response values, missingness, or an analysis method. These gaps matter because incomplete follow-up can affect both the denominator and the interpretation of observed changes over time. No assumption about the completeness of follow-up or the treatment of missing data is warranted from the posted information.

The lack of reported multiplicity and missing-data methods is not evidence that no methods were used; it means they are not reported in the registry record. The results available for interpretation are limited to the posted safety summaries.

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 5% reporting threshold.

Most frequent non-serious adverse event terms

The safety summary records serious adverse events as 0 of 3 at risk in the 100 μg TriMix cohort, 0 of 3 in the 300 μg TriMix cohort, 0 of 3 in the 600μg cohort, 0 of 6 in the 900μg cohort, and 0 of 6 in the 1200μg cohort. Deaths are recorded as 0 in each cohort. No serious adverse-event terms are listed.

The separate other-adverse-event counts are 3 of 3, 3 of 3, 2 of 3, 4 of 6 and 3 of 6 across those cohorts, respectively. These figures indicate participants affected in that category as reported; they do not state the number of individual events. A participant may have had an event that does not meet the primary endpoint definition, and the summary does not provide enough detail to classify events as DLTs.

General statistical principle: zero recorded serious events does not establish zero risk. The study cohorts are small, and the registry record does not give uncertainty intervals or detailed follow-up completeness for these summaries. The counts describe what the posted results record, not a guarantee about outcomes beyond the study.

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

The posted results describe adverse-event counts by sequential dose cohort. They show 0 serious adverse events recorded in each cohort, 0 deaths in each cohort, and the reported numbers of participants with other adverse events. This is the scope of the safety information that can be stated directly from the registry record.

These observations do not establish that iHIVARNA-01 is safe or unsafe, and they do not show that one dose is safer than another. The study was not randomized, the cohorts were enrolled sequentially, and the registry does not report the primary DLT result. The other-adverse-event summaries cannot stand in for DLT because the endpoint definition includes severity, confirmation, attribution and discontinuation criteria not captured by those counts.

The record also does not report results for the listed immunogenicity or reservoir outcomes. It therefore does not support conclusions about changes in HIV-1-specific immune response, reservoir effects, or treatment benefit. The stated aim of evaluating safety and establishing a recommended dose is not itself evidence that a recommended dose was established.

General statistical principle: a small descriptive dataset can identify what was observed in the enrolled cohorts, while leaving considerable uncertainty about less common events and about comparisons between regimens. The absence of a reported event in these cohorts should not be interpreted as evidence that the event is impossible. The registry’s posted information also does not provide enough detail to judge the completeness of follow-up or how missing assessments might affect results.

Thus, the defensible reading is narrow: the record gives descriptive safety summaries for the listed cohorts, but not the primary endpoint results or analyses needed to answer the study’s principal dose-limiting toxicity question. No broader efficacy or comparative claim follows from the posted counts.

14. Why This Trial Matters Statistically

This study illustrates a common statistical challenge in early-phase dose escalation: the design is organized around sequential cohorts and safety monitoring, while the posted summary may contain only limited outcome detail. The cohort structure and escalation rules help explain the intended decision process, but they do not provide the actual DLT outcomes or establish which dose was recommended.

It also shows why endpoint definitions matter. A count of participants with any other adverse event is not interchangeable with a count meeting the DLT definition. For interpretation, the event criteria, assessment window, denominator and participant-level classification all matter. Without the primary endpoint results, the study’s central safety question cannot be assessed from the registry record.

The non-randomized design is another important feature. Sequential cohorts can be useful for structured dose evaluation, but their counts are descriptive and are not a substitute for a randomized comparison. Cohort differences may reflect factors other than dose, and the record does not provide analyses that address such differences.

Finally, the study lists secondary immune-response and reservoir outcomes, but no results or analysis methods for them are reported. This distinction between outcomes listed in a registry and outcomes with interpretable posted results is important when reviewing early-phase research. The record permits a careful description of the design and the adverse-event summaries, while also making clear how much remains unreported.

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.

The main limitation is that the registry record does not report the primary DLT counts, despite identifying DLT by week 24 as the primary endpoint. The serious and other adverse-event summaries cannot fill that gap because their categories are not equivalent to the protocol’s DLT definition. The record also does not give event-level details, severity distributions, attribution, or discontinuations by cohort.

The study was open label, non-randomized and sequential. It therefore does not provide a like-for-like randomized comparison across dose groups. Small cohort sizes also limit the precision of descriptive safety estimates, and the registry does not report confidence intervals for the posted counts. The record does not state how complete follow-up was or how missing safety assessments were handled.

The intervention descriptions contain an inconsistency for the 300 μg TriMix cohort: the arm title gives 300 μg, while the arm description says 100 μg of TriMix mRNA. The registry record does not resolve this. It also does not report the results for immunogenicity or reservoir outcomes, the analysis populations, or the methods for repeated measurements and multiplicity.

Finally, the posted record does not report a sample-size justification, interim analyses, or a full statistical analysis plan. These omissions restrict assessment of the study’s precision and analysis choices. They should be read as limitations of what is reported in the registry record, not as proof that the study team did not address these issues elsewhere.

16. Related Tutorials

17. Related Statistical Calculators

18. Sources

• ClinicalTrials.gov: Phase I, Open Label Dose Escalation Study to Evaluate Safety of iHIVARNA-01 in Chronically HIV-infected Patients — NCT02413645.
• Registry record last updated 2025-08-22; status last verified 2025-08.
• Results first posted 2025-08-22.
• 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

NCT02413645 is a completed Phase I, open-label, non-randomized, sequential dose-escalation study of iHIVARNA-01 in chronically HIV-infected patients under stable combined antiretroviral therapy. The registry reports enrollment of 21 and lists five dose cohorts. The primary endpoint was DLT through week 24, defined by specified severe adverse events and vaccination-attributable discontinuation.

The posted safety summaries record 0 serious adverse events and 0 deaths in each cohort, alongside cohort-specific counts for participants with other adverse events. The registry record does not report the primary DLT results, inferential analyses, or results for the listed immunogenicity and reservoir outcomes. Consequently, the available record supports descriptive reporting of the posted safety categories, but not a comparative dose conclusion or a conclusion about immune response or reservoir effects.