Marcussen Mette, Bødker Julie Støve, Christensen Heidi Søgaard, Johansen Preben, Nielsen Søren, Christiansen Ilse, Bergmann Olav Jonas, Bøgsted Martin, Dybkær Karen, Vyberg Mogens, Johnsen Hans Erik
Department of Clinical Medicine, Aalborg University, Sdr. Skovvej 15, Aalborg, Denmark.
Clinical Cancer Research Center, Aalborg University Hospital, Sdr. Skovvej 15, Aalborg, Denmark.
PLoS One. 2017 Jan 4;12(1):e0169286. doi: 10.1371/journal.pone.0169286. eCollection 2017.
Toxicity of the oral and gastrointestinal mucosa induced by high-dose melphalan is a clinical challenge with no documented prophylactic interventions or predictive tests. The aim of this study was to describe molecular changes in human oral mucosa and to identify biomarkers correlated with the grade of clinical mucositis.
Ten patients with multiple myeloma (MM) were included. For each patient, we acquired three buccal biopsies, one before, one at 2 days, and one at 20 days after high-dose melphalan administration. We also acquired buccal biopsies from 10 healthy individuals that served as controls. We analyzed the biopsies for global gene expression and performed an immunohistochemical analysis to determine HLA-DRB5 expression. We evaluated associations between clinical mucositis and gene expression profiles. Compared to gene expression levels before and 20 days after therapy, at two days after melphalan treatment, we found gene regulation in the p53 and TNF pathways (MDM2, INPPD5, TIGAR), which favored anti-apoptotic defense, and upregulation of immunoregulatory genes (TREM2, LAMP3) in mucosal dendritic cells. This upregulation was independent of clinical mucositis. HLA-DRB1 and HLA-DRB5 (surface receptors on dendritic cells) were expressed at low levels in all patients with MM, in the subgroup of patients with ulcerative mucositis (UM), and in controls; in contrast, the subgroup with low-grade mucositis (NM) displayed 5-6 fold increases in HLA-DRB1 and HLA-DRB5 expression in the first two biopsies, independent of melphalan treatment. Moreover, different splice variants of HLA-DRB1 were expressed in the UM and NM subgroups.
Our results revealed that, among patients with MM, immunoregulatory genes and genes involved in defense against apoptosis were affected immediately after melphalan administration, independent of the presence of clinical mucositis. Furthermore, our results suggested that the expression levels of HLA-DRB1 and HLA-DRB5 may serve as potential predictive biomarkers for mucositis severity.
高剂量美法仑引起的口腔和胃肠道黏膜毒性是一项临床挑战，目前尚无已记录的预防性干预措施或预测性检测方法。本研究的目的是描述人类口腔黏膜的分子变化，并确定与临床黏膜炎分级相关的生物标志物。
纳入10例多发性骨髓瘤（MM）患者。对于每位患者，我们在高剂量美法仑给药前、给药后2天和给药后20天分别采集三次颊黏膜活检样本。我们还从10名健康个体获取颊黏膜活检样本作为对照。我们分析活检样本的全基因表达情况，并进行免疫组织化学分析以确定HLA-DRB5的表达。我们评估了临床黏膜炎与基因表达谱之间的关联。与治疗前和治疗后20天的基因表达水平相比，在美法仑治疗后2天，我们发现p53和TNF通路（MDM2、INPPD5、TIGAR）中的基因调控有利于抗凋亡防御，并且黏膜树突状细胞中的免疫调节基因（TREM2、LAMP3）上调。这种上调与临床黏膜炎无关。HLA-DRB1和HLA-DRB5（树突状细胞表面受体）在所有MM患者、溃疡性黏膜炎（UM）患者亚组以及对照组中均低表达；相比之下，轻度黏膜炎（NM）亚组在前两次活检中HLA-DRB1和HLA-DRB5表达增加了5 - 6倍，与美法仑治疗无关。此外，UM和NM亚组中表达了不同的HLA-DRB1剪接变体。
我们的结果显示，在MM患者中，美法仑给药后免疫调节基因和参与抗凋亡防御的基因立即受到影响，与临床黏膜炎的存在无关。此外，我们的结果表明HLA-DRB1和HLA-DRB5的表达水平可能作为黏膜炎严重程度的潜在预测生物标志物。
