Department of Dermatology and Department of Medicine, Division of Rheumatology, Brigham and Women's Hospital, Boston, Massachusetts, USA.
Harvard Medical School, Boston, Massachusetts, USA.
Lupus Sci Med. 2021 Mar;8(1). doi: 10.1136/lupus-2020-000428.
Punch biopsy, a standard diagnostic procedure for patients with cutaneous lupus erythematosus (CLE) carries an infection risk, is invasive, uncomfortable and potentially scarring, and impedes patient recruitment in clinical trials. Non-invasive tape sampling is an alternative that could enable serial evaluation of specific lesions. This cross-sectional pilot research study evaluated the use of a non-invasive adhesive tape device to collect messenger RNA (mRNA) from the skin surface of participants with CLE and healthy volunteers (HVs) and investigated its feasibility to detect biologically meaningful differences between samples collected from participants with CLE and samples from HVs.
Affected and unaffected skin tape samples and simultaneous punch biopsies were collected from 10 participants with CLE. Unaffected skin tape and punch biopsies were collected from 10 HVs. Paired samples were tested using quantitative PCR for a candidate immune gene panel and semi-quantitative immunohistochemistry for hallmark CLE proteins.
mRNA collected using the tape device was of sufficient quality for amplification of 94 candidate immune genes. Among these, we found an interferon (IFN)-dominant gene cluster that differentiated CLE-affected from HV (23-fold change; p<0.001) and CLE-unaffected skin (sevenfold change; p=0.002), respectively. We found a CLE-associated gene cluster that differentiated CLE-affected from HV (fourfold change; p=0.005) and CLE-unaffected skin (fourfold change; p=0.012), respectively. Spearman's correlation between per cent area myxovirus 1 protein immunoreactivity and IFN-dominant mRNA gene cluster expression was highly significant (dermis, rho=0.86, p<0.001). In total, skin tape-derived RNA expression comprising both IFN-dominant and CLE-associated gene clusters correlated with per cent area immunoreactivity of some hallmark CLE-associated proteins in punch biopsies from the same lesions.
A non-invasive tape RNA collection technique is a potential tool for repeated skin biomarker measures throughout a clinical trial.
对于患有皮肤红斑狼疮 (CLE) 的患者,钻孔活检是一种标准的诊断程序,但具有感染风险、侵袭性、不适和潜在的疤痕形成,这会妨碍临床试验中患者的招募。非侵入性的胶带采样是一种替代方法,可以对特定病变进行连续评估。这项横断面试点研究评估了使用非侵入性的胶带设备从 CLE 患者和健康志愿者 (HV) 的皮肤表面采集信使 RNA (mRNA) 的情况,并研究了其从 CLE 患者采集的样本和 HV 样本之间检测到生物学上有意义的差异的可行性。
从 10 名 CLE 患者中采集了受影响和未受影响的皮肤胶带样本和同时的钻孔活检样本。从 10 名 HV 中采集了未受影响的皮肤胶带和钻孔活检样本。使用定量 PCR 对候选免疫基因进行了测试,对免疫组化半定量检测了 CLE 标志性蛋白。
使用胶带设备采集的 mRNA 质量足以扩增 94 个候选免疫基因。在这些基因中,我们发现了一个干扰素 (IFN)-主导的基因簇,可以区分 CLE 受影响和 HV(23 倍变化;p<0.001)和 CLE 未受影响的皮肤(七倍变化;p=0.002)。我们发现了一个 CLE 相关的基因簇,可以区分 CLE 受影响和 HV(四倍变化;p=0.005)和 CLE 未受影响的皮肤(四倍变化;p=0.012)。在真皮中,Myxovirus 1 蛋白免疫反应的百分比与 IFN-主导的 mRNA 基因簇表达之间的 Spearman 相关性非常显著(rho=0.86,p<0.001)。总的来说,皮肤胶带衍生的 RNA 表达包括 IFN-主导和 CLE 相关基因簇,与来自同一病变的钻孔活检中一些标志性 CLE 相关蛋白的免疫反应百分比相关。
非侵入性的胶带 RNA 采集技术是在临床试验中重复进行皮肤生物标志物测量的潜在工具。