神经母细胞瘤的精准治疗研究进展
Advances in precision therapy research for neuroblastoma

中山大学肿瘤防治中心
Sun Yat-sen University Cancer Center

张翼鷟
Zhang Yixing river gull

2024.12.14 福州
2024.12.14 Fuzhou

目录
catalogs

CONTENT

劳拉替尼在ALK基因驱动高危复发/难治NB中的应用
Loratinib in ALK gene-driven high-risk relapsed/refractory NB

达妥昔单抗β联合PD-1抗体在初诊高危复发/难治NB中的应用
Daltuximab beta in combination with PD-1 antibody in primary diagnosis of high-risk relapsed/refractory NB

NK细胞输注治疗NB的转化与临床研究
Translational and Clinical Studies of NK Cell Infusion for the Treatment of NB

表观遗传调控在NB发病机制中作用的一项基础研究
A basic study of the role of epigenetic regulation in the pathogenesis of NB

劳拉替尼在ALK基因驱动高危复发/难治NB中的应用
Loratinib in ALK gene-driven high-risk relapsed/refractory NB

劳拉替尼在ALK基因驱动高危复发/难治NB中的应用 N=17
Loratinib in ALK gene-driven high-risk relapsed/refractory NB N=17

2023年8月至2024年8月，共17例NB患者接受劳拉替尼单药或联合治疗，中位年龄6岁1-46岁；
Between August 2023 and August 2024, a total of 17 NB patients with a median age of 6 years1-46 years were treated with loratinib monotherapy or combination therapy;

截止2024年12月，11例患者仍在服药，3例因PD停药，3例因经济原因停药，中位服药时间6月2-13月
As of December 2024, 11 patients were still taking the drug, 3 were discontinued due to PD, and 3 were discontinued due to financial reasons, with a median time on drug of 2-13 months in June

劳拉替尼在ALK基因驱动高危复发/难治NB中的疗效分析
Efficacy analysis of loratinib in ALK gene-driven high-risk relapsed/refractory NB

劳拉替尼联合化疗在ALK基因扩增和突变患者中均显示较好疗效
Loratinib Combination Chemotherapy Shows Better Efficacy in Both ALK Gene Amplification and Mutation Patients

少见ALK突变或ALK胚系突变可能对劳拉替尼耐药
Rare ALK mutations or ALK germline mutations may be resistant to loratinib

劳拉替尼联合诱导化疗有效率达到100%
Loratinib Combination Induction Chemotherapy Achieves 100% Efficacy Rate

同时伴有MYCN扩增的患者接受劳拉替尼治疗的有效率较低
Patients with concomitant MYCN amplification treated with loratinib had lower efficacy rates

外周血ctDNA动态监测能较好反应临床疗效
Peripheral blood ctDNA ambulatory monitoring better reflects clinical outcomes

劳拉替尼单药或联合化疗安全性良好，常见不良反应包括食欲增加高三油血症等
Loratinib is safe as a single agent or in combination with chemotherapy, with common adverse effects including increased appetite hypertriglyceridemia

案例： 劳拉替尼联合化疗成功挽救ALK基因驱动复发性高危NB2012
Case: Successful rescue of ALK gene-driven recurrent high-risk NB2012 by combination chemotherapy with loratinib

一般资料
General information

男15岁
Male 15 years old

初治诊断：腹膜后神经母细胞瘤M期 高危组 MYCN-
Primary diagnosis: retroperitoneal neuroblastoma stage M High-risk group MYCN-

确诊时间：2012年3月
Diagnosed: March 2012

2012年
2012

初治诊断：腹膜后神经母细胞瘤M期 高危组 MYCN-
Primary diagnosis: retroperitoneal neuroblastoma stage M High-risk group MYCN-

CAV/IE/IC交替化疗10程+手术，疗效CR
CAV/IE/IC 10 courses of alternating chemotherapy + surgery, efficacy CR

维甲酸+干扰素维持1年
Retinoic acid + interferon maintenance for 1 year

2014

停治疗10个月，第一次复发： 骨髓复发
10 months off treatment, first relapse: bone marrow relapse

VIT方案*4程SD
VIT Program *4 Program SD

VIP方案*6程CR
VIP Program *6 Program CR

CT：+NVB口服节拍化疗1年
CT: + NVB oral beat chemotherapy for 1 year

2023

停治疗7年，第二次复发：多发骨 骨髓复发
Stopped treatment for 7 years, second relapse: multiple bones Bone marrow relapse

入组临床研究：AIT*6程，骨髓CR，，颅脑MRI怀疑脑膜复发 改行
Enrolled in clinical study: course of AIT*6, bone marrow CR,,, suspected meningeal recurrence on cranial MRI Revisited

CT：+TOPO方案*2程
CT: + TOPO program * 2 programs

MIBG评 分0分，颅脑MRI未见病灶，获得CR
MIBG score 0, no lesions seen on cranial MRI, CR obtained

2024年5月MIBG平分16分，第三次复发
May 2024 MIBG tie score of 16, third relapse

第三次复发后辅助检查
Ancillary tests after the third relapse

2024年5月复查提示 多发骨 右侧顶部硬脑膜复发
May 2024 review suggests multiple bones, right parietal dural recurrence.

全脊髓MRI未见脊髓转移
No spinal cord metastases seen on whole spinal cord MRI

123I-MIBG评分：16分 2,2,2,2,2,2,0,2,2,0
123I-MIBG score: 16 2,2,2,2,2,2,2,0,2,2,0

双部位骨髓涂片均未见肿瘤浸润
Bone marrow smears from both sites showed no tumor infiltration

颅脑MRI：右侧顶部硬脑膜局限增厚，考虑转移
Cranial MRI: limited thickening of the right parietal dura, considering metastasis

初诊肿瘤组织和复发时外周血NGS检测
Peripheral blood NGS testing at initial tumor tissue diagnosis and at recurrence

患者初诊时肿瘤组织和复发时外周血NGS检测结果
Patient's tumor tissue at initial diagnosis and peripheral blood NGS test results at recurrence

初诊时肿瘤组织中无ALK基因突变
No ALK gene mutation in tumor tissue at initial diagnosis

复发时外周血中测到ALK基因突变， 可匹配到劳拉替尼
ALK mutation detected in peripheral blood at relapse, matched to loratinib

第三次复发后的治疗
Treatment after the third relapse

2024-5-22至2024-9-3行CE方案(CBP+VP-16)联合劳拉替尼(150mg po QD)治疗6程
6 courses of CE regimen (CBP + VP-16) in combination with loratinib (150 mg po QD) from 2024-5-22 to 2024-9-3

4程后复查MIBG L1 L5局部MIBG可疑弱阳性(CRu) Curie评分2分(0,0,0,2,0,0,0,0,0,0)
Review of MIBG L1 L5 after 4 courses Localized MIBG suspiciously weakly positive (CRu) Curie score 2 (0,0,0,2,0,0,0,0,0,0)

第4程后外周血ctDNA基因数明显减少，峰度下降
The number of peripheral blood ctDNA genes decreased significantly after course 4, with a decrease in kurtosis

第4程后颅脑MRI原右侧顶部硬脑膜增厚消失
Disappearance of the original right parietal dural thickening on cranial MRI after the 4th procedure

第6程后外周血ctDNA清零
Peripheral blood ctDNA clearance after course 6

第6程后双部位骨穿未见肿瘤
No tumor was seen on double-site bone puncture after the 6th procedure

第6程后行L1 L5和右侧顶部硬脑膜TOMO放疗30G，/15f
L1 L5 and right parietal dural TOMO radiotherapy after course 6 30G,/15f

目前已行8程CE联合劳拉替尼治疗(末次化疗时间2024-11-12)
Currently on 8 courses of CE in combination with Loratinib (last chemotherapy 2024-11-12)

计划全面复查(MIBG 颅脑MRI 骨穿 ctDNA) 再决定下一步治疗……
A full review is planned (MIBG cranial MRI bone puncture ctDNA) before deciding on the next step in treatment ......

第三次复发后的治疗后疗效
Post-treatment outcome after the third relapse

治疗前 4程后治疗前 治疗前 4程后 6程后
Pre-treatment 4 post-treatment Pre-treatment 4 post-treatment 6 post-treatment

达妥昔单抗β联合PD-1抗体在初诊高危复发/难治NB中的应用
Daltuximab beta in combination with PD-1 antibody in primary diagnosis of high-risk relapsed/refractory NB

达妥昔单抗β联合PD-1抗体在初诊高危复发/难治NB中的应用 N=5
Daltuximab beta in combination with PD-1 antibody in primary high-risk relapsed/refractory NB N=5

2023年9月至2024年7月，N=5，包括：2例初诊高危NB，3例难治性高危NB；
September 2023 to July 2024, N=5, including: 2 primary high-risk NB, 3 refractory high-risk NB;

中位年龄5岁1-5岁，中位5程3-9程；
Median age 5 years 1-5 years, median 5 courses 3-9 courses;

疗效：3例无靶病灶患者持续CR，2例存在靶病灶疗效均SD；
Efficacy: 3 patients with no target lesions sustained CR and 2 with target lesions present efficacy were SD;

生存：从应用GD2单抗开始，截止2024年12月，中位PFS时间10个月5-13月，5例患者均存活；
SURVIVAL: Starting with the application of GD2 monoclonal antibody and ending in December 2024, with a median PFS time of 10 months 5-13 months, all 5 patients survived;

达妥昔单抗β不良反应：包括发热疼痛水肿皮疹等，轻微可控；
Daltuximab beta adverse reactions: including fever pain edema rash, etc., mild and controllable;

PD-1抗体不良反应：1例患者出现I度肝功能异常先甲亢后甲减甲亢时无心率快等症状
Adverse reactions to PD-1 antibody: 1 patient developed I degree liver function abnormality first hyperthyroidism and then hypothyroidism Hyperthyroidism without fast heart rate and other symptoms

案例：达妥昔单抗β联合PD-1抗体维持治疗改善难治性NB患儿生 存和提高生活质量
Case: Daltuximab Beta Combined with PD-1 Antibody Maintenance Therapy Improves Survival and Quality of Life for Children with Refractory NB

病史资料
medical history

基本情况：男, 3岁2月
Basic: Male, 3 years and 2 months old

主诉：低热腹痛下肢痛半月余
Complaint: low-grade fever, abdominal pain, lower extremity pain for more than half a month.

现病史：因无明显诱因间断低热腹痛左下肢痛 ，就诊于当地医院，行 CT检查发现右侧肾上腺区及邻近腹膜后见多发结节状肿块状软组织密度影，范围约28*64*57mm，与邻近肝实质分界不清，考虑神经母细胸瘤并周围转移瘤 2023年4月25日就诊于我院
Current medical history: due to intermittent low-grade fever and abdominal pain without obvious triggers, left lower extremity pain, consultation in the local hospital, CT examination found that the right adrenal region and adjacent retroperitoneum, multiple nodular mass-like soft tissue density shadow, the range of about 28 * 64 * 57mm, and the adjacent liver parenchyma demarcation is not clear, consider neuroblastoma and peripheral metastases on April 25, 2023, in our hospital

既往史：无特殊
Past history: none specific

个人史：无特殊
Personal history: none in particular

家族史：无特殊
Family history: none specific

辅助检查结果
Ancillary findings

血常规：WBC 5.6*109/L，HB 104g/L，PLT 484*109/L
Blood routine: WBC 5.6*109/L, HB 104g/L, PLT 484*109/L

NSE：216ng/mL
NSE: 216ng/mL

尿VMA：2.29mmol/ml正常＜14.9mmol/ml
Urine VMA: 2.29 mmol/ml normal <14.9 mmol/ml

尿HVA：15.1mmol/ml 正常＜28.8mol/ml
Urine HVA: 15.1 mmol/ml normal <28.8 mol/ml

病理右肾上腺肿物：神经母细胞瘤，分化差型
Pathologic right adrenal mass: neuroblastoma, poorly differentiated type

辅助检查结果
Ancillary findings

PET-CT示：右侧肾上腺病灶代谢活跃，考虑神经母细胞瘤，约3.0*3.1cm，病灶侵犯邻近肝组织：右侧腹膜后多发淋巴结代谢活跃，考虑转移：全身多发骨病灶代谢活跃，考虑转移，部分病灶侵犯双侧邻近胸膜
PET-CT: right adrenal lesion metabolically active, consider neuroblastoma, about 3.0*3.1cm, lesion invades adjacent liver tissue: right retroperitoneal multiple lymph nodes metabolically active, consider metastasis: systemic multiple bone lesions metabolically active, consider metastasis, part of the lesion invades adjacent pleura bilaterally

颅脑MRI：颅脑未见占位病变
Cranial MRI: no space-occupying lesion in the cranium

左后髂骨骨髓涂片：涂片偶见成堆分布的肿瘤细胞
Bone marrow smear of the left posterior ilium: occasional mounds of tumor cells in the smear

左后髂骨流式：0.165%的细胞表达GD2+CD56+CD81+
Left posterior iliac flow-through: 0.165% of cells expressing GD2+CD56+CD81+

胸骨骨髓涂片：涂片可见大量恶性肿瘤细胞
Bone marrow smear of sternum: large number of malignant tumor cells seen in the smear

胸骨流式：21.92%的细胞表达GD2+CD56+CD81+
Sternal flow: 21.92% of cells expressed GD2+CD56+CD81+

分子生物学检查
Molecular Biology

右肾上腺肿瘤组织标本WES结果示：
WES results of right adrenal tumor tissue specimen are shown:

染色体11q缺失，BRAF突变，BIRC5扩增4，RHBDF2扩增，RPTOR扩增，SO：9扩增，SPHK1扩增，SRSF2扩增ALK阴性，MYCN 3.2拷贝考虑获得状态
Chromosome 11q deletion, BRAF mutation, BIRC5 amplification 4, RHBDF2 amplification, RPTOR amplification, SO:9 amplification, SPHK1 amplification, SRSF2 amplification ALK-negative, MYCN 3.2 copies considered for acquisition of status

PDL122C3：CPS 10，
PDL122C3: CPS 10.

TMB-L：0.6 Muts/Mb
TMB-L: 0.6 Muts/Mb

MSI：MSS
MSI: MSS

MYCN扩增阴性
MYCN amplification negative

ALK突变阴性
ALK mutation negative

临床诊断及危险度分层参考NCCN指南 2024.01
Refer to NCCN guidelines for clinical diagnosis and risk stratification 2024.01

临床诊断：右肾上腺神经母细胞瘤 M期 高危组 分化差型 MYCN阴性 11q缺失
Clinical diagnosis: right adrenal neuroblastoma, stage M, high-risk group, poorly differentiated, MYCN-negative, 11q-deficient

治疗过程总结
Summary of the treatment process

确诊
make a definite diagnosis

诱导化疗C1-3
Induction chemotherapy C1-3

2022-8-20至 2022-10-01 行CAV/VIP交替治疗
2022-8-20 to 2022-10-01 Perform CAV/VIP alternating therapy

2程化疗后骨髓CR、原发肿瘤PR
Bone marrow CR, primary tumor PR after 2 courses of chemotherapy

手术
surgeries

右侧后腹膜肿瘤切除+右侧肾上腺切除+右肾固定+后腹膜淋巴结清扫术
Right retroperitoneal tumor resection + right adrenalectomy + right kidney fixation + retroperitoneal lymph node dissection

C4

2022-11-04行VIP(IFO0.9g+DDP15mg+VP16 0.06g)
2022-11-04 line VIP (IFO 0.9g + DDP 15mg + VP16 0.06g)

术后复查CT：未见肿瘤残留；骨髓持续CR；ECT示：全身多发骨代谢仍较活跃
Postoperative follow-up CT: no tumor residue seen; bone marrow persistent CR; ECT: systemic multiple bone metabolism still active

诱导治疗C5-8
Induction therapy C5-8

2022-11-25 至2023-1-30 行CAV/VIP交替治疗
2022-11-25 to 2023-1-30 Perform CAV/VIP alternating therapy

诱导治疗后：L4代 谢活跃，考虑肿瘤 残留；骨髓CR 总疗效PR，定为难治性
After induction therapy: L4 metabolism is active, consider residual tumor; bone marrow CR total efficacy PR, classified as refractory

桥接治疗 C1-2
Bridging therapy C1-2

2023-2-28至2023-3-18 行阿帕替尼+IT方案
2023-2-28 to 2023-3-18 Perform Apatinib + IT regimen

2程IT方案后123I-M -IB结果提示骨转 移，Curie评分：12 分(2,3,2,2,2,0,01,0)
123I-M-IB results suggestive of bone metastasis after 2 courses of IT program, Curie score: 12 (2,3,2,2,2,2,0,01,0)

挽救化疗共6线方案
Rescue chemotherapy total 6 line regimen

包括：ICE HITS+PD-1抗体 多美素+奈达铂 CT：+TOPO多 美素+奈达铂+达 妥昔单抗β DDP+VM-26 方 案
Includes: ICE HITS+PD-1 antibody Dormin+Nedaplatin CT: +TOPO Dormin+Nedaplatin+Datuximab beta DDP+VM-26 Programs

123I-MIBG显像阳 性，提示骨转移， Curie评分14-15 分，疗效SD。
Positive 123I-MIBG image suggests bone metastasis, Curie score 14-15, efficacy SD.

多线挽救化疗后 疗效SD，2024 年1月至今行达 妥昔单抗β+PD -1抗体维持治疗， 目前已9程
SD after multiple lines of salvage chemotherapy, maintenance therapy with dalcolizumab beta + PD-1 antibody from January 2024 to present, currently on 9 courses

定期复查123IMIBG检查， Curie评分15分左 右，疗效持续SD， 患儿目前无症状， 一般情况良好
Regularly reviewed 123 IMIBG, Curie score of 15, persistent SD, child is asymptomatic and in good general condition.

达妥昔单抗β联合PD-1抗体治疗期间疗效评估
Evaluation of efficacy during treatment with daltuximab beta in combination with PD-1 antibodies

2024年1月至2024年12月，行达妥昔单抗β联合纳武利尤单抗9程，定期复查MIBG，疗效SD
9 courses of daltuximab beta in combination with natalizumab from January 2024 to December 2024, with regular follow-up MIBG, efficacy SD

患者应用那西妥单抗β维持治疗前常诉下肢痛，维持治疗后，下肢疼痛症状消失，肿瘤稳定控制11个月，生活质量良好
The patient often complained of lower limb pain before applying nacitumumab beta maintenance therapy, and after the maintenance therapy, the lower limb pain symptoms disappeared, and the tumor has been under stable control for 11 months, with a good quality of life

治疗前：123I-MIBG检查Curie评分15分
Prior to treatment: 123I-MIBG exam Curie score 15

5轮治疗后：123I-MIBG检查Curie评分15分
After 5 rounds of treatment: 123I-MIBG exam Curie score 15

NK细胞输注治疗NB的转化与临床研究
Translational and Clinical Studies of NK Cell Infusion for the Treatment of NB

临床问题：NK细胞治疗可否应用于神经母细胞瘤
Clinical question: can NK cell therapy be applied to neuroblastoma?

神经母细胞瘤：冷肿瘤
Neuroblastoma: a cold tumor

NK细胞输注在NB中开展的前瞻性临床研究
Prospective clinical study of NK cell infusion performed in NB

PD-L1+NK细胞预测神经母细胞瘤预后及PD-1/PD-L1抗体疗效
PD-L1+NK cells predict neuroblastoma prognosis and PD-1/PD-L1 antibody efficacy

NK细胞浸润预示良好预后
NK cell infiltration predicts good prognosis

PD-L1+NK细胞处于免疫激活状态
PD-L1+ NK cells in an immune-activated state

PD-L1+NK细胞预测神经母细胞瘤预后及PD-1/PD-L1抗体疗效
PD-L1+NK cells predict neuroblastoma prognosis and PD-1/PD-L1 antibody efficacy

PD-L1+NK细胞比例高、功能强
High proportion of PD-L1+ NK cells with high function

PD-L1+NK细胞浸润预示良好预后
PD-L1+ NK cell infiltration predicts good prognosis

PD-L1+NK细胞预测神经母细胞瘤预后及PD-1/PD-L1抗体疗效
PD-L1+NK cells predict neuroblastoma prognosis and PD-1/PD-L1 antibody efficacy

体内外实验验证阻断PD-1/PD-L1增强NK细胞对神经母细胞瘤的杀伤作用
In vitro and in vivo experiments validate that blocking PD-1/PD-L1 enhances the killing effect of NK cells on neuroblastoma

脐带血NK细胞联合GD2抗体在神经母细胞瘤中发挥强效作用
Umbilical cord blood NK cells combined with GD2 antibody exert potent effects in neuroblastoma

神经母细胞瘤GD2抗体治疗获得CR的患者，NK细胞浸润的比例更高，PFS更佳
Patients who achieved CR on neuroblastoma GD2 antibody therapy had a higher percentage of NK cell infiltration and better PFS

脐带血NK细胞联合GD2抗体在神经母细胞瘤中发挥强效作用
Umbilical cord blood NK cells combined with GD2 antibody exert potent effects in neuroblastoma

相比外周血，脐带血NK细胞联合GD2抗体在体内抗肿瘤作用更强，显著改善生存
Cord blood NK cells combined with GD2 antibody have stronger anti-tumor effects in vivo compared to peripheral blood, significantly improving survival

脐带血NK细胞联合GD2抗体在神经母细胞瘤中发挥强效作用
Umbilical cord blood NK cells combined with GD2 antibody exert potent effects in neuroblastoma

相比外周血，脐带血NK细胞明显活化，细胞毒性更强，体外联合GD2抗体杀伤作用更强
Compared to peripheral blood, cord blood NK cells were significantly activated, more cytotoxic, and more potent in vitro combined with GD2 antibody killing

脐带血NK细胞联合GD2抗体在神经母细胞瘤中发挥强效作用
Umbilical cord blood NK cells combined with GD2 antibody exert potent effects in neuroblastoma

脐带血NK细胞回输在小鼠模型中安全性可控，未出现明显毒性
Umbilical cord blood NK cell transfusion has a manageable safety profile in a mouse model without significant toxicity

一项脐带血自然杀伤(NK)细胞治疗儿童高危复发/难治性神经母细胞瘤的I期临床研究
A Phase I Clinical Study of Umbilical Cord Blood Natural Killer (NK) Cells for the Treatment of High-Risk Relapsed/Refractory Neuroblastoma in Children

伦理获批
Ethics approved

招募患者
Recruitment of patients

NK细胞准备
NK cell preparation

NK细胞输注
NK cell infusion

中山大学肿瘤防治中心体细胞临床研究开展情况
Development of clinical research on somatic cells at Sun Yat-sen University Cancer Center

全国首批国家卫健委体细胞临床研究机构资质
Qualification of the First National Health Commission Body Cell Clinical Research Institutes

制定体细胞临床研究质量管理相关制度和SOP
Development of systems and SOPs related to quality management of somatic cell clinical studies

建立体细胞临床研究全流程数字化平台
Establishment of a digital platform for the whole process of somatic cell clinical research

密闭系统生产： C 级背景下的细胞生产系统
Closed System Production: Cell Production Systems in a Class C Context

达妥昔单抗β联合脐血NK细胞在高危难治/复发NB中的应用 N=4
Daltuximab beta combined with cord blood NK cells in high-risk refractory/relapsed NB N=4

2024年11月7日至2024年12月12日，4例难治性NB患者接受达妥昔单抗β联合脐血NK细胞输注；
Between November 7, 2024 and December 12, 2024, four patients with refractory NB received daltuximab beta in combination with cord blood NK cell infusion;

中位年龄5岁（4-5岁），中位既往化疗方案线数3线（2-7线）
Median age 5 years (4-5 years), median number of lines of prior chemotherapy regimens 3 lines (2-7 lines)

3例存在靶病灶，1例无靶病灶；
Target lesions were present in 3 cases and absent in 1 case;

4例患者均尚未评估疗效；
Efficacy has not been assessed in any of the 4 patients;

4例患者脐血NK细胞输注过程中未出现不良反应，安全性良好
No adverse reactions occurred during cord blood NK cell infusion in 4 patients, with a favorable safety profile

案例：
Case in point:

病史资料
medical history

基本情况：
The basics:

姓名：李××
Name：Li××

性别：男
Sex: Male

诊断时年龄：3岁
Age at diagnosis: 3 years

诊断：腹膜后神经母细胞瘤伴骨髓多发骨转移 M期 高危 难治性 NM，C基因扩增阴性
Diagnosis: retroperitoneal neuroblastoma with multiple bone metastases in the bone marrow Stage M High-risk Refractory NM with negative C gene amplification

一线治疗
front-line treatment

2023.2至 2023.6
2023.2 to 2023.6

行三氧化二砷+CAV/EP 交替化疗C1-C4
Arsenic trioxide + CAV/EP alternating chemotherapy C1-C4

软组织病灶疗效：2程后4程后：SD； 骨髓疗效：2程后4程后骨髓：CR
Soft tissue lesion efficacy: after 2 courses after 4 courses: SD; Bone marrow efficacy: after 2 courses after 4 courses bone marrow: CR

2023.7.7腹膜后肿瘤切除术
2023.7.7 Resection of retroperitoneal tumors

二倍体核型 SCA-
Diploid karyotype SCA-

2023.7至 2023.10
2023.7 to 2023.10

行三氧化二砷+EP化疗C5
Performed arsenic trioxide + EP chemotherapy C5

行CAV化疗C6
CAV chemotherapy C6

行EP化疗C7
EP chemotherapy C7

行CT化疗C8-C9
Perform CT chemotherapy C8-C9

2023.12原发灶放疗
2023.12 Radiotherapy to primary site

总疗效：PR 残留S1及左侧胫骨局部病灶
Overall outcome: PR Residual S1 and localized left tibial lesions

2021.1

全面复查
full review

MIBG：0分
MIBG: 0 points

PET/CT：残留
PET/CT: residual

2024-3

入组甲磺酸阿帕替尼联合IT方案治疗复发难治儿童神经母细胞瘤的多中心单臂II期临床研究 行AIT阿帕替
Enrollment in a multicenter, single-arm, phase II clinical study of apatinib mesylate in combination with an IT regimen for the treatment of relapsed, refractory pediatric neuroblastoma Line AIT apatinib

尼+伊立替康+替莫唑胺方案化疗C1-5；
Ni + irinotecan + temozolomide regimen for chemotherapy C1-5;

复查PET/CT：疗效CR
Repeat PET/CT: efficacy CR

2024.7

2024.7.10 行自体造血干细胞移植
2024.7.10 Autologous Hematopoietic Stem Cell Transplantation

2024-10

全面复查：CR
Comprehensive review: CR

行达妥昔单抗β联合GM-CSF免疫治疗C1-C2
Performed daltuximab beta combined with GM-CSF immunotherapy C1-C2

2024-11

行脐带血NK细胞输注C1
Perform cord blood NK cell infusion C1

继续达妥昔单抗β联合GM-CSF联合NK细胞治疗中
Continued daltuximab beta in combination with GM-CSF in combination with NK cell therapy

表观遗传调控在NB发病机制中作用的一项基础研究
A basic study of the role of epigenetic regulation in the pathogenesis of NB

研究背景
Background of the study

高危神经母细胞瘤NB患者预后差，生存率低，缺乏有效治疗手段，表观遗传疗法可能是一种有前途的替代方案
Patients with high-risk neuroblastoma NB have a poor prognosis, low survival rates, and lack of effective treatments; epigenetic therapy may be a promising alternative

SUV39H1是一种含有SET结构域的组蛋白甲基转移酶，负责组蛋白H3上第9位赖氨酸K9 的3甲基化H3K9me3 ，以H3K9me3依赖一种表观遗传修饰方式的方式，介导对靶基因的转录抑制
SUV39H1 is a histone methyltransferase containing a SET structural domain responsible for the 3-methylation of lysine K9 at position 9 on histone H3 H3K9me3 , which mediates transcriptional repression of target genes in a manner that is dependent on an epigenetic modification of H3K9me3

研究背景
Background of the study

越来越多的研究表明，SUV39H1在许多肿瘤中表达上调，包括人类结肠癌，膀胱癌和肝细胞癌，黑色素瘤等，并与肿瘤的增殖，侵袭，迁移，凋亡抵抗及预后相关，发挥促肿瘤作用，目前, SUV39H1在神经母细胞瘤NB中的作用及调控机制尚处于未知
More and more studies have shown that SUV39H1 is up-regulated in many tumors, including human colon, bladder and hepatocellular carcinomas, melanoma, etc., and is associated with tumor proliferation, invasion, migration, apoptosis resistance and prognosis, and plays a pro-tumorigenic role, Currently, the roles of SUV39H1 and its regulatory mechanisms in neuroblastoma NB are still unknown.

chaetocin毛壳素，一种从真菌中分离出来的天然代谢产物，是SUV39H1的高选择性抑制剂，具有多种生物活性和药理功能，在黑色素瘤，非小细胞肺癌，卵巢癌，胃癌，胶质瘤，肝癌，乳腺癌等多种肿瘤中发挥抗肿瘤的作用，但其在NB中的作用及机制尚未有研究
chaetocin trichothecene, a natural metabolite isolated from fungi, is a highly selective inhibitor of SUV39H1 with a variety of bioactivities and pharmacological functions, exerting antitumor effects in a variety of tumors, including melanoma, NSCLC, ovarian cancer, gastric cancer, glioma, hepatocellular carcinoma, and breast cancer; however, its role in NB and the mechanism have not yet been investigated

技术路线
technological route

NB细胞系中小分子药库高通量药筛
High-throughput drug screen for small and medium molecule drug libraries in NB cell lines

chaetocin药效显著（毛壳素：SUV39H1高选择抑制剂）
Chaetocin is highly potent (trichothecenes: highly selective inhibitor of SUV39H1)

体外细胞系及小鼠体内明确SUV39H1的促肿瘤功能
Defining the tumor-promoting function of SUV39H1 in an in vitro cell line and in vivo in mice

体外细胞系及小鼠体内验证毛壳素药效
In vitro cell line and in vivo mouse validation of hairy chitin efficacy

RNA-seq转录组 测 序 寻 找 下游 调 控 的 靶 基因MCPIP1及被显 著 改 变 的 周期通路
RNA-seq transcriptome sequencing to identify downstream-regulated target gene MCPIP1 and significantly altered cyclic pathways

体外细胞系中 明 确MCPIP1的抑癌功能
Confirmation of the oncogenic function of MCPIP1 in in vitro cell lines

明确SUV39H1 通过H3K9三甲 基化沉默 MCPIP1，影响 下游周期通路 的调节机制
Defining the regulatory mechanism of SUV39H1 that affects the downstream cycling pathway by silencing MCPIP1 through H3K9 trimethylation

研究结果1. 高通量药物筛选发现SUV39H1抑制剂毛壳素具有显 著抗NB作用
Findings 1. High-throughput drug screening reveals that the SUV39H1 inhibitor trichothecene has a significant anti-NB effect

A-B: 小分子药库筛选，发现毛壳素对NB细胞系具有显著杀伤作用；
A-B: Screening of a small molecule drug library revealed significant killing effects of trichothecenes on NB cell lines;

C-D: 数据库生信分析发现SUV39H1在NB中高表达，与NB患者不良预后相关；
C-D: Database raw letter analysis revealed that SUV39H1 is highly expressed in NB and is associated with poor prognosis in NB patients;

E-I: NB细胞系及本中心NB肿瘤组织中验证SUV39H1高表达，与NB患者不良预后相关；
E-I: High expression of SUV39H1 was verified in NB cell lines and NB tumor tissues from our center and correlated with poor prognosis in NB patients;

研究结果2. 毛壳素在体外和体内对NB细胞均显示抗肿瘤作用
Findings 2. Trichothecenes show anti-tumor effects on NB cells in vitro and in vivo

A-H: 体外功能实验表明毛壳素的抗NB作用：抑制NB细胞增殖，迁移，促进细胞凋亡，介导细胞周期阻滞在G2期；
A-H: In vitro functional assays demonstrated the anti-NB effects of trichothecenes: inhibition of NB cell proliferation, migration, promotion of apoptosis, and mediation of cell cycle arrest in G2 phase;

I-M: 小鼠体内实验显示毛壳素的抗NB作用：抑制肿瘤细胞在小鼠体内的生长；
I-M: In vivo experiments in mice show the anti-NB effect of trichothecenes: inhibition of tumor cell growth in mice;

研究结果3. SUV39H1在体外和体内均显示对NB细胞的促肿瘤作用
Finding 3. SUV39H1 shows tumor-promoting effects on NB cells in vitro and in vivo

A-H: 体外功能实验表明SUV39H1的促NB作用：促进NB细胞增殖，迁移，抑制细胞凋亡，敲低SUV39H1，可介导细胞周期阻滞在G2期；
A-H: In vitro functional assays demonstrated the pro-NB effects of SUV39H1: promoted NB cell proliferation, migration, inhibited apoptosis, and knockdown of SUV39H1 mediated cell cycle arrest in G2 phase;

I-M: 小鼠体内实验表明SUV39H1的促NB作用：促进NB肿瘤细胞在小鼠体内的生长；
I-M: In vivo experiments in mice demonstrated the pro-NB effect of SUV39H1: promotion of NB tumor cell growth in mice;

研究结果4. SUV39H1在NB中激活细胞周期通路
Finding 4. SUV39H1 activates cell cycle pathway in NB

A-B: RNA-seq测序结果表明，在NB细胞系中抑制SUV39H1后，细胞周期通路显著改变；
A-B: RNA-seq sequencing results showed that cell cycle pathways were significantly altered upon inhibition of SUV39H1 in NB cell lines;

C-F: 在NB细胞系中，抑制SUV39H1 (chaetocin/shRNA敲低)后，G2期周期转换蛋白显著变化；
C-F: Significant changes in G2 phase cycle transition proteins after inhibition of SUV39H1 (chaetocin/shRNA knockdown) in NB cell lines;

G: SUV39H1与AURKA呈正相关；
G: SUV39H1 was positively correlated with AURKA;

H: 周期回复实验表明, SUV39H1对NB细胞的周期调控作用，依赖于AURKA的表达；
H: Cycle-response experiments indicate that the cycle-regulatory effect of SUV39H1 on NB cells is dependent on AURKA expression;

（AURKA是SUV39H1对NB周期调控的关键分子）
(AURKA is a key molecule in the regulation of the NB cycle by SUV39H1)

研究结果5. MCPIP1是SUV39H1的潜在下游调节因子
Finding 5. MCPIP1 is a potential downstream regulator of SUV39H1

A-B: RNA-seq测序结果表明，在NB细胞系中抑制SUV39H1后，细胞周期通路显著改变；
A-B: RNA-seq sequencing results showed that cell cycle pathways were significantly altered upon inhibition of SUV39H1 in NB cell lines;

A-D: 在NB细胞系中，抑制SUV39H1 (chaetocin/shRNA敲低)后，MCPIP1显著上调；
A-D: MCPIP1 was significantly upregulated after inhibition of SUV39H1 (chaetocin/shRNA knockdown) in NB cell lines;

E: SUV39H1与MCPIP1呈正相关；
E: SUV39H1 is positively correlated with MCPIP1;

F-M: MCPIP1在NB中低表达，具有抑制NB细胞生长的作用；
F-M: MCPIP1 is lowly expressed in NB and has a role in inhibiting NB cell growth;

研究结果6. MCPIP1在NB中下调AURKA的表达
Finding 6. MCPIP1 downregulates AURKA expression in NB

A-B: 在NB细胞系中，过表达MCPIP1后，AURKA显著上调；
A-B: AURKA was significantly upregulated after overexpression of MCPIP1 in NB cell lines;

C-F: 过表达AURKA后，可以逆转由过表达MCPIP1引起的对NB增殖的抑制效应；
C-F: Overexpression of AURKA reversed the inhibitory effect on NB proliferation induced by overexpression of MCPIP1;

MCPIP1在NB中下调AURKA,且其对NB细胞的抑制作用，依赖于其对AURKA的表达调控作用；
MCPIP1 downregulates AURKA in NB, and its inhibitory effect on NB cells is dependent on its role in regulating AURKA expression;

研究结果7. SUV39H1通过MCPIP1-AURKA信号轴调控NB细胞的细胞周期进程
Finding 7. SUV39H1 regulates cell cycle progression in NB cells through the MCPIP1-AURKA signaling axis

A-F: CHIP-PCR及回复实验表明：SUV39H1在NB中，通过MCPIP1-AURKA信号轴发挥促肿瘤作用
A-F: CHIP-PCR and replication experiments show that SUV39H1 exerts a pro-tumorigenic effect through the MCPIP1-AURKA signaling axis in NB

总结
summarize

SUV39H1在NB细胞中显著高表达并起促肿瘤作用；
SUV39H1 is significantly overexpressed in NB cells and plays a tumor-promoting role;

从机制上讲，NB细胞中高表达的SUV39H1通过表观遗传修饰转录沉默其靶基因MCPIP1具体表现为：SUV39H1使H3K9me3在MCPIP1启动子区域的富集增加，进而减少MCPIP1介导的AURKA降解；导致AURKA累积增多，促进细胞周期进程，引起NB肿瘤恶性进展；
Mechanistically, SUV39H1, which is highly expressed in NB cells, transcriptionally silences its target gene MCPIP1 through epigenetic modification as follows: SUV39H1 increases the enrichment of H3K9me3 in the promoter region of MCPIP1, which in turn reduces the MCPIP1-mediated degradation of AURKA; leading to increased accumulation of AURKA, which promotes cell cycle progression and causes NB tumor malignant progression;

(SUV39H1-MCPIP1-AURKA调控轴)
(SUV39H1-MCPIP1-AURKA regulation axis)

毛壳素可以抑制SUV39H1的H3K9甲基转移酶活性以及NB细胞周期进展，可作为一种有效，安全的表观遗传疗法，对NB患者产生潜在临床益处
Trichothecenes inhibit H3K9 methyltransferase activity of SUV39H1 and NB cell cycle progression, and could be an effective, safe epigenetic therapy with potential clinical benefit for NB patients

谢谢 Q&A
Thanks Q&A