Background:Graft versus host disease (GVHD) and the graft versus tumor (GVT) effect after allogeneic hematopoietic cell transplantation (allo-HCT) result from complex interactions between the donor immune system and the recipient environment. High-temporal longitudinal monitoring might be necessary to identify triggering events of GVHD and GVT and to intercept these events before their occurrence. But it would require an overall considerable amount of blood by venipuncture, which is unfeasible in such a fragile population.Methods:In this study, we implemented a targeted multiplex microfluidics q-PCR-based transcriptional fingerprint assay (TFA) on 50 µL of blood collected by a simple fingerstick to evaluate post-allo-HCT systemic immune perturbations associated with the development of GVHD. Fluctuations of a panel of 264 genes were measured in 31 allo-HCT patients by frequent (weekly or biweekly) analysis of 50 µL serial blood samples. Cross-sectional and longitudinal analyses correlated with detailed clinical annotations were performed.Results:Signatures of neutrophil activation and interferon (IFN) characterized the onset of acute GVHD, while an ongoing cytotoxic response was modulated in chronic mild GVHD and protein-synthesis and B-cell-related signatures characterized late acute/overlap GVHD. An unexpected erythroid signature distinguished patients with acute and mild chronic GVHD.Conclusions:Our micro-invasive approach unveiled the molecular heterogeneity of GVHD and identified hierarchically important biological processes conducive to different forms of GVHD. These findings increase our understanding of GVHD and reveal potentially targetable alterations. This approach might be implemented clinically to intercept GVHD before its occurrence and to modulate therapeutic interventions accordingly.
背景：异基因造血细胞移植（allo-HCT）后的移植物抗宿主病（GVHD）与移植物抗肿瘤（GVT）效应源于供体免疫系统与受体环境之间复杂的相互作用。为识别GVHD和GVT的触发事件并在其发生前进行干预，可能需要进行高时间分辨度的纵向监测。但这通常需要通过静脉穿刺采集大量血液，对于此类脆弱患者群体而言并不可行。 方法：本研究采用基于靶向多重微流控定量PCR的转录指纹分析（TFA），仅需通过简单指尖采血获取50微升血液，即可评估与GVHD发生相关的allo-HCT后系统性免疫紊乱。通过对31例allo-HCT患者每周或每两周采集的50微升系列血样进行分析，监测了264个基因的表达波动，并结合详细的临床注释进行横断面与纵向关联分析。 结果：中性粒细胞激活和干扰素（IFN）相关特征标志急性GVHD的发作；慢性轻度GVHD中呈现持续受调节的细胞毒性反应；而蛋白质合成与B细胞相关特征则表征晚期急性/重叠型GVHD。研究意外发现，红细胞相关特征可区分急性与轻度慢性GVHD患者。 结论：本研究的微创检测方法揭示了GVHD的分子异质性，并识别出导致不同形式GVHD的关键层级化生物学过程。这些发现深化了我们对GVHD的理解，并揭示了潜在的可靶向干预机制。该方法未来或可应用于临床，在GVHD发生前进行干预，并据此调整治疗策略。
Whole-Blood Longitudinal Molecular Profiling Maps the Road of Graft Versus Host Disease (GVHD)
肿瘤(癌症)患者之家