OMIC-11. SINGLE CELL RNA SEQUENCING FROM THE CSF OF SUBJECTS WITH H3K27M+ DIPG/DMG TREATED WITH GD2 CAR T-CELLULAR THERAPY
OMIC-11. SINGLE CELL RNA SEQUENCING FROM THE CSF OF SUBJECTS WITH H3K27M+ DIPG/DMG TREATED WITH GD2 CAR T-CELLULAR THERAPY
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DOI：
10.1093/neuonc/noab090.158
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发表时间：
2021-06-01
期刊：
影响因子：
15.9
通讯作者：
Monje M
中科院分区：
文献类型：
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作者：
Mochizuki A;Ramakrishna S;Good Z;Patel S;Chinnasamy H;Yeom K;Schultz L;Richards R;Campen C;Reschke A;Mahdi J;Toland A;Baggot C;Mavroukakis S;Egeler E;Moon J;Landrum K;Erickson C;Rasmussen L;Barsan V;Tamaresis J;Marcy A;Kunicki M;Celones M;Ehlinger Z;Kurra S;Cornell T;Partap S;Fisher P;Grant G;Vogel H;Davis K;Feldman S;Sahaf B;Majzner R;Mackall C;Monje M
我们正在进行一项I期临床试验，利用靶向GD 2的嵌合抗原受体（CAR）T细胞（NCT 04196413）治疗H3 K27 M突变型弥漫性内在脑桥胶质瘤（DIPG）和脊髓弥漫性中线胶质瘤（DMG）。在通过Ommaya导管进行常规颅内压监测时采集脑脊液（CSF）用于相关研究。在这里，我们展示了前3名受试者的单细胞RNA测序结果。 利用10 X Genomics对在CAR T细胞施用之前和之后的各个时间点从CSF分离的细胞和CAR T细胞产物进行单细胞RNA测序。将输出与Cell Ranger对齐，并在R中进行分析。 如Majzner等人在本次会议上提交的摘要所述，在剂量水平1治疗的4例受试者中，有3例显示出明确的影像学和/或临床获益。迄今为止，我们已经完成了这四个受试者中三个受试者的单细胞RNA测序（两个受益，一个没有）。在过滤掉低质量信号和双联体后，分析了3名受试者的89，604个细胞。其中，4，122个细胞代表从CSF分离的细胞，85，482个细胞代表CAR T细胞产物。与未获得治疗应答的受试者相比，2例表现出明显临床和影像学改善的受试者在输注前CSF中表现出更少的S100 A8 + S100 A9+骨髓抑制细胞和CD 25 + FOXP 3+调节性T细胞。在一名表现出改善的患有DIPG的受试者中，通过轨迹分析，在第+14天在CSF中可检测到的多克隆CAR T细胞与输注前CAR T细胞相比表现出CD 8A、GZMA、GNLY和PDCD 1的富集，表明向细胞毒性表型分化;同一受试者随着时间的推移表现出S100 A8 + S100 A9+骨髓细胞和CX 3CR 1 + P2 RY 12+小胶质细胞的数量增加。将在获得数据后提供进一步分析。 在CSF中可检测到的免疫抑制性骨髓群体的存在可能与DIPG/DMG的CAR T细胞疗法中的临床应答相关。
We are conducting a Phase I clinical trial utilizing chimeric antigen receptor (CAR) T-cells targeting GD2 (NCT04196413) for H3K27M-mutant diffuse intrinsic pontine glioma (DIPG) and spinal cord diffuse midline glioma (DMG). Cerebrospinal fluid (CSF) is collected for correlative studies at the time of routine intracranial pressure monitoring via Ommaya catheter. Here we present single cell RNA-sequencing results from the first 3 subjects. Single cell RNA-sequencing was performed utilizing 10X Genomics on cells isolated from CSF at various time points before and after CAR T-cell administration and on the CAR T-cell product. Output was aligned with Cell Ranger and analyzed in R. As detailed in the Majzner et al. abstract presented at this meeting, three of four subjects treated at dose-level one exhibited clear radiographic and/or clinical benefit. We have to date completed single cell RNA-sequencing for three of these four subjects (two with benefit, one without). After filtering out low-quality signals and doublets, 89,604 cells across 3 subjects were analyzed. Of these, 4,122 cells represent cells isolated from CSF and 85,482 cells represent CAR T-cell product. Two subjects who demonstrated clear clinical and radiographic improvement exhibited fewer S100A8+S100A9+ myeloid suppressor-cells and CD25+FOXP3+ regulatory T-cells in the CSF pre-infusion compared to the subject who did not derive a therapeutic response. In one subject with DIPG who demonstrated improvement, polyclonal CAR T-cells detectable in CSF at Day +14 demonstrated enrichment of CD8A, GZMA, GNLY and PDCD1 compared to the pre-infusion CAR T-cells by trajectory analysis, suggesting differentiation toward a cytotoxic phenotype; the same subject exhibited increasing numbers of S100A8+S100A9+ myeloid cells and CX3CR1+P2RY12+ microglia over time. Further analyses will be presented as data become available. The presence of immunosuppressive myeloid populations, detectable in CSF, may correlate to clinical response in CAR T cell therapy for DIPG/DMG.