Safety and Feasibility of FMT for Immune Recovery After HSCT

Overview

Patients who undergo allogeneic hematopoietic stem cell transplantation (allo-HSCT) often experience slow and incomplete recovery of their immune system, particularly the part responsible for producing antibodies (humoral immunity). This can lead to increased risk of infections and other complications. Previous studies suggest that the gut microbiome and its metabolites, such as short-chain fatty acids (propionate and butyrate), play a critical role in immune recovery. This study has two phases. In the first phase, we will observe changes in immune function, lymphocyte subsets, and gut metabolites (propionate and butyrate) before and at 1, 3, and 6 months after transplantation in 50-80 patients. We will compare patients with severe immune deficiency to those with normal recovery to identify differences in gut bacteria and metabolites. In the second phase, we will perform selective fecal microbiota transplantation (FMT) in 6 patients who still have severe humoral immunodeficiency at 6 months post-transplant. The FMT capsules will be specially selected from donors whose gut bacteria are rich in both propionate-producing and butyrate-producing strains. Patients will receive 20 capsules daily for 3 consecutive days (60 capsules total). We will evaluate the safety and feasibility of this approach, and observe whether fecal secretory immunoglobulin A (sIgA) shows signs of recovery at 3 months after FMT. The study is expected to take approximately 2 years to complete. Findings may provide a new precision microbiome-based strategy to promote immune recovery after transplantation.

BACKGROUND Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is a curative treatment for various hematological malignancies. However, B-cell-mediated humoral immune reconstitution after allo-HSCT is remarkably slow and often incomplete. Immunoglobulin M (IgM) typically recovers within 2-6 months, immunoglobulin G (IgG) within 3-18 months, while immunoglobulin A (IgA) recovery may be delayed up to 3 years or longer. Functional humoral immune recovery often requires more than 5 years. Previous studies have shown that acute and chronic graft-versus-host disease (GVHD) are significant contributors to persistent IgA deficiency. The conditioning regimen prior to transplantation causes profound disruption of the gut microbiota, impairing the bidirectional regulatory network between the microbiome and host immunity. Microbial metabolites, particularly short-chain fatty acids (SCFAs) such as propionate and butyrate, play essential roles in B-cell differentiation and antibody production. Propionate is critical for inducing and maintaining intestinal regulatory T cells (Tregs), while butyrate provides energy for B-cell differentiation and immunoglobulin secretion. Preliminary clinical data from the investigators' group show that 100% (5/5) of immunodeficient patients post-allo-HSCT exhibit specific propionate deficiency, whereas patients with general dysbiosis primarily show butyrate deficiency. This suggests that the propionate synthesis pathway, driven predominantly by Bacteroidetes, is more severely destroyed under the extreme post-transplant environment. Based on ecological keystone species theory, propionate-producing bacteria (particularly Bacteroides species) are highly connected nodes in the gut microbial network. A cross-feeding relationship exists between propionate- and butyrate-producing bacteria, where the former degrade complex carbohydrates to provide growth substrates for the latter. The investigators propose the "propionate first, butyrate follows" two-stage synergistic reconstruction hypothesis: Stage 1 (Microenvironment Preparation): Propionate-producing bacteria are preferentially restored, increasing intestinal propionate levels, inducing Treg cells, and creating an immune-permissive environment. Stage 2 (Effector Activation): Butyrate-producing bacteria successfully colonize in the improved microenvironment, restoring butyrate synthesis and driving B-cell differentiation and sIgA secretion. A prior Phase II trial by Rashidi and colleagues (2023) demonstrated that oral fecal microbiota transplantation (FMT) is safe and feasible in allo-HSCT patients and may reduce GVHD risk. Based on this evidence and the novel hypothesis, the investigators propose an innovative selective FMT strategy using a "super donor" matching approach. Study Design This is a two-phase, single-center study conducted at The First Affiliated Hospital of Soochow University. Phase 1 (Prospective Observational Cohort): Allo-HSCT patients are enrolled and followed pre-transplant, and at 1, 3, and 6 months post-transplant. At each visit, peripheral blood and fecal samples are collected for serum immunoglobulins, fecal sIgA, TBNK lymphocyte subsets, fecal 16S rRNA sequencing, quantitative SCFAs (propionate and butyrate), and dietary assessment. At 6 months post-transplant, patients are stratified into three groups based on serum IgA levels: immunodeficiency (IgA \<0.07 g/L with IgG/IgM not recovered), delayed recovery (0.07 g/L ≤ IgA ≤ 0.8 g/L, or IgA \<0.07 but IgG/IgM recovered), and normal recovery (IgA \>0.8 g/L with IgG/IgM normal). Extreme phenotype analysis (immunodeficiency vs. normal recovery) will be used for microbiota and metabolite comparisons. Phase 2 (Single-arm, Open-label, Phase I/IIa Proof-of-concept Safety Trial): Patients from the immunodeficiency group at 6 months post-transplant who meet additional inclusion criteria are enrolled. A baseline comprehensive assessment is performed to map the microbiota-metabolite deficiency profile, followed by matching with a donor whose gut bacteria are rich in both propionate-producing and butyrate-producing strains. Participants receive oral FMT capsules (≥2×10¹¹ organisms/g), 20 capsules daily for 3 consecutive days (total 60 capsules per course). Follow-up visits occur at 2 weeks, 1 month, 2 months, and 3 months post-FMT, with immune parameters, microbiome profiling, SCFAs, and dietary assessment repeated at each visit. Safety Monitoring Safety is monitored at each visit using CTCAE v5.0 criteria. Serious adverse events are reported to the Principal Investigator and Ethics Committee within 24 hours. Strict donor screening, exclusion of active GVHD, and emergency preparedness are implemented. If Grade 3 or higher FMT-related adverse events occur, treatment is suspended and a safety review is initiated. Statistical Methods Phase 1 analyses include linear mixed-effects models for longitudinal trends, LEfSe for differential taxa screening, and random forest/LASSO regression for IgA prediction modeling. Phase 2 endpoints are analyzed using Wilcoxon signed-rank tests for paired comparisons and Spearman correlation for temporal associations. Missing data are handled using last observation carried forward. Two-sided tests with a significance level of 0.05 will be used. All analyses will be performed using R software (version 4.x or later).