Genomic and Phenotypic Determinants of Resistance to Immunotherapies in Multiple Myeloma
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 40
- 试验地点
- 1
- 主要终点
- Biomarkers of response
研究概览
简要总结
A total of 40 Multiple Myeloma (MM) patients at clinical relapse who progressed during Proteasome Inhibitors (PIs) or Immunomodulating Drugs (IMiDs)-based therapies and who are assigned to antiCD38-based salvage treatments, will be enrolled. We will collect bone marrow (BM) and peripheral blood (PB) samples from patients at specific timepoints:
- baseline (BM, PB and buccal swab)
- every 3 month (PB)
- achievement of response (≥ Very Good Partial Response (VGPR)) (BM and PB)
- relapse or refractory status to antiCD38-based treatments (BM and PB) Samples will be processed and stored in the "Hematological Laboratory" located in the University of Turin (Italy) for various proposed analyses: at specific time-points CD138+ (Plasma Cells-PCs) and marker CD138/19+ (B cells) will be immunomagnetically enriched from the BM mononuclear cells and frozen as viable cells in dimethyl sulfoxide (DMSO); PB mononuclear cells (PBMCs) will be isolated from whole blood by density-gradient centrifugation, and frozen as above; plasma fraction from PB and BM will be obtained by centrifugation and stored frozen; a buccal swab will be obtained at the time of enrollment as a source of control germline DNA and stored frozen.
详细描述
Aim 1: Evaluation of cell-intrinsic mechanisms on BM.
- Whole Genome Sequencing (WGS) and Whole Exome Sequencing (WES) will be performed on marker CD138+ purified cells to evaluate their genomic profile, and on buccal swab DNA to restrict the analysis to variants or structural abnormalities that have a clear somatic status, and are therefore specific to the tumor cells. In details:
1.1 WGS: libraries will be prepared with TruSeq™(Kit Illumina) DNA Polymerase Chain Reaction (PCR)-Free Library Preparation Kit (Illumina, San Diego, CA) from 500ng of genomic DNA, aiming for an average target insert of 300bp. Sequencing will be performed on a 150bp-paired end protocol, at a target depth of 40x for tumor samples and 30x for normal samples.
1.2 WES: libraries will be prepared with SureSelectXT Human All Exon V6 (Agilent technologies int., Santa Clara, CA) from 100ng of genomic DNA, aiming for an average target insert of 300bp. Sequencing will be performed on a 150bp-paired end protocol, aiming for a target depth of 200x for tumor samples and 100x for normal samples.
1.3 Data analysis: next generation sequencing scoring system output format files (*.FASTQ files) will be aligned to the reference genome using Burrows-Wheeler Alignment Tool (BWAmem), and deduplicated aligned Binary Alignment Map (BAM) files will be analyzed using the following published tools available at the Wellcome Trust Sanger Institute (WTSI):
- accurate genome-wide allele-specific copy number (ASCAT) and Battenberg for clonal and subclonal copy number changes.
- BRASS for structural variations (large inversions and deletions, translocations, internal tandem duplication).
- Caveman and Pindel for Single Nucleotide Variants (SNVs) and small insertion-deletions (indels).
研究设计
- 研究类型
- Observational
- 观察模型
- Other
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •MM patients at clinical relapse who progressed during PIs or IMiDs-based therapies
- •Patients assigned to antiCD38-based salvage treatments
- •Patients with measurable disease
排除标准
- •No criteria
结局指标
主要结局
Biomarkers of response
时间窗: 5 years
Response measured through mutational analysis. Results will be integrated by statistical models (e.g. univariate, multivariate analysis) and then correlated to cytofluorimetric results and patients' available clinical data in order to predict outcome.
Cell extrinsic mechanisms of response
时间窗: 5 years
The analysis will include T-cells population (eg. CD38+, CD4+, CD8+, Tregs cells), regulatory and suppressive immune populations (MDSCs) and cytokines (eg Activin-A, IL-3, IL-6, RANKL, OPG, MIP-1α, MIP-3α and DKK-1) characterization.
Sensitivity vs resistance to new immunotherapies
时间窗: 5 years
The patients' response to new drugs administration will be evaluated identifying genomic aberrations (e.g. TRAF3 deletion/mutation and Cereblon mutation) or impaired cellular surface molecules expression (eg CD38, CD55, CD59 expression).
次要结局
未报告次要终点
研究者
Mario Boccadoro
Principal Investigator
University of Turin, Italy