CN117607442B - 一种预测乳腺癌免疫治疗效果的标志物、试剂盒与应用 - Google Patents
一种预测乳腺癌免疫治疗效果的标志物、试剂盒与应用 Download PDFInfo
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Abstract
本发明属于生物医药领域,具体涉及一种预测乳腺癌免疫治疗效果的标志物、试剂盒与应用。更具体地,本发明涉及一种预测乳腺癌免疫治疗效果的标志物,其中所述乳腺癌是HER2阳性乳腺癌或HER2低表达乳腺癌,所述免疫治疗是抗HER2抗体治疗,并且所述生物标志物是SHCBP1。本发明还涉及用于检测SHCBP1表达的试剂包含其的试剂盒及其用于预测乳腺癌免疫治疗效果的用途。本发明还涉及抑制SHCBP1表达的试剂用于增强抗HER2抗体治疗乳腺癌的效果的用途。本发明已发现SHCBP1的表达水平与抗HER2抗体治疗乳腺癌的效果相关,因而可以用于预测抗HER2抗体治疗乳腺癌的效果。本发明还进一步发现抑制SHCBP1的表达水平可以提升抗HER2抗体治疗乳腺癌尤其是HER2低表达乳腺癌的效果。
Description
技术领域
本发明属于生物医药领域,具体涉及一种预测乳腺癌免疫治疗效果的标志物、试剂盒与应用。
背景技术
HER2又称人表皮生长因子受体-2,是一种原癌基因,在细胞膜上表达HER2蛋白,当EGF与HER-2结合后,引起HER2由单体变成二聚体,随后引发自体磷酸化而传递细胞内信息,调控基因表达,最后维持正常的细胞生长与分裂。当HER2表达过多,细胞会因过度刺激而造成不正常的快速生长,最终造成癌症发生。
HER2阳性乳腺癌是恶性程度高的一类乳腺癌分型,乳腺癌HER2阳性大约占全部乳腺癌病例的15-20%。抗HER2抗体,例如曲妥珠单抗。是能与HER2特异结合的单克隆抗体,能专一性与HER2结合,阻止EGF与HER2结合,抑制癌细胞生长。另外,曲妥珠单抗还能降低癌细胞HER2的表达,抑制肿瘤血管新生的作用,诱发体内免疫作用,包括抗体依赖型细胞毒杀作用与补体毒杀作用,进而造成被单克隆抗体辨识的目标细胞死亡,从而发挥抗肿瘤的作用。
然而抗HER2抗体的治疗并非对所有的HER2阳性乳腺癌患者均有效,例如近30%的HER2阳性乳腺癌患者在使用曲妥珠单抗治疗的过程中对曲妥珠单抗产生耐药,从而无法曲妥珠单抗方案的治疗中获益。
此外,随着ADC药物的飞速发展,HER2低表达的定义被提出,即HER2 IHC 1+或2+且无HER2扩增。如果按比例划分的话,真正检测不到HER2的乳腺癌,在全部乳腺癌中大约占30-40%,而剩下45-55%左右的乳腺癌,就是所谓“HER2低表达乳腺癌”。
现有技术中存在针对乳腺癌的靶点研究,赵诗哲等于2020年发表在《生物信息学》上的研究论文“SHCBP1:乳腺癌治疗候选靶点和潜在预后标志物”揭示了利用SHCBP1作为乳腺癌治疗候选靶点的研究成果,但未针对HER2低表达乳腺癌进行研究。
兰州大学第二医院焦作义团队2021年在Nature Communications上发表的题为“Hyperactivation of HER2-SHCBP1-PLK1 axis promotes tumor cell mitosis andimpairs trastuzumab sensitivity to gastric cancer”的研究论文,发现了HER2下游存在一条新的信号通路,但同样缺乏针对HER2低表达乳腺癌的研究。
而抗HER2抗体在HER2低表达乳腺癌中的治疗效果存在显著差异。例如临床研究表明,曲妥珠单抗在HER2低表达乳腺癌中的治疗效果有限,而德卢替康-曲妥珠单抗(抗体-药物缀合物DS-8201)则显示良好的治疗效果。
因此,本领域急需可以判断乳腺癌尤其是HER2阳性乳腺癌和HER2低表达乳腺癌患者的治疗效果的标志物。
发明内容
为了解决上述问题,在第一方面,本发明提供了一种预测乳腺癌免疫治疗效果的标志物,其中所述乳腺癌是HER2阳性乳腺癌或HER2低表达乳腺癌,所述免疫治疗是抗HER2抗体治疗,并且所述生物标志物是SHCBP1。
进一步地,相较于正常组织,乳腺癌组织中较高水平的SHCBP1表达指示较差的乳腺癌免疫治疗效果;相较于正常组织,乳腺癌组织中较低水平或相似水平的SHCBP1表达指示较好的乳腺癌免疫治疗效果。
如本文所用,术语“HER2阳性乳腺癌”意指在免疫组织化学(IHC)中HER2表达评分为3+的乳腺癌,和在IHC中HER2表达评分为2+并在原位杂交(ISH)中HER2表达被确定为阳性的乳腺癌。所述原位杂交方法包括荧光原位杂交方法(FISH)和双色原位杂交方法(DISH)。
如本文所用,术语“HER2低表达乳腺癌”没有特别限制,只要它被本领域技术人员认为是低表达HER2的乳腺癌。HER2低表达乳腺癌的优选实例可包括在IHC法中HER2表达评分为2+且在原位杂交法中HER2表达被确定为阴性的乳腺癌,以及在IHC法中HER2表达评分为1+的乳腺癌。
如本文所用,术语“抗HER2抗体”是指与HER2(人表皮生长因子受体2型;ErbB-2)特异性结合的抗体,并且优选具有通过与HER2结合而在表达HER2的细胞中内化的活性。抗HER2抗体的具体实例可以包括曲妥珠单抗及其缀合物例如德卢替康-曲妥珠单抗(DS-8201)
在第二方面,本发明提供了一种预测乳腺癌免疫治疗效果的试剂盒,其中所述试剂盒包含检测SHCBP1表达的试剂,其中所述乳腺癌是HER2阳性乳腺癌或HER2低表达乳腺癌,所述免疫治疗是抗HER2抗体治疗。
进一步地,相较于正常组织,乳腺癌组织中较高水平的SHCBP1表达指示较差的乳腺癌免疫治疗效果;相较于正常组织,乳腺癌组织中较低水平或相似水平的SHCBP1表达指示较好的乳腺癌免疫治疗效果。
在第三方面,本发明提供了检测SHCBP1表达的试剂在制备用于预测乳腺癌免疫治疗效果的工具中的用途,其中所述乳腺癌是HER2阳性乳腺癌或HER2低表达乳腺癌,所述免疫治疗是抗HER2抗体治疗。
进一步地,相较于正常组织,乳腺癌组织中较高水平的SHCBP1表达指示较差的乳腺癌免疫治疗效果;相较于正常组织,乳腺癌组织中较低水平或相似水平的SHCBP1表达指示较好的乳腺癌免疫治疗效果。
如本文所用,“检测SHCBP1表达的试剂”没有特别限制,只要其能够检测SHCBP1蛋白或mRNA的表达量。例如,检测SHCBP1表达的试剂可以包括以下方法中使用的试剂:蛋白质印迹(Western Blot法)、酶联免疫吸附测定(ELISA)、放射性免疫测定(RIA)、夹心测定、免疫组织化学染色、质谱法、免疫沉淀分析法、补体结合分析法、流式细胞荧光分边技术和蛋白质芯片法。
在第四方面,本发明提供了抑制SHCBP1表达的试剂在制备用于增强抗HER2抗体治疗乳腺癌的效果的药物中的用途,其特征在于,所述乳腺癌是HER2低表达乳腺癌。
进一步地,所述抗HER2抗体选自曲妥珠单抗或德卢替康-曲妥珠单抗。
进一步地,所述抑制SHCBP1表达的试剂包括抑制SHCBP1表达的核酸分子、碳水化合物、脂、小分子化合物、抗体、多肽、蛋白、基因编辑载体、慢病毒或腺相关病毒中的一种或多种。
进一步地,所述抑制SHCBP1表达可通过本领域技术人员所熟知的基因突变、基因沉默、基因敲除、基因编辑或基因敲减技术实现。如利用RNA干扰(RNAi)技术可以特异性剔除或关闭特定基因的表达;利用基因编辑技术的工具可以是CRISPR/Cas9技术、锌指核酸酶(ZFNs)或转录激活因子样效应物核酸酶(TALENs)技术等,但不限于此。利用基因敲减(基因敲低)技术从转录后水平或翻译水平使ADAR1基因表达失活或基因沉默的技术为本领域技术人员所熟知。所述基因敲减技术包括RNA干扰、Morpholino干扰、反义核酸、核酶或显性负抑制突变但不限于此。利用病毒(如慢病毒、腺相关病毒)表达的shRNA或siRNA抑制基因的表达,进行基因的沉默是本领域技术人员所熟知的。
进一步地,所述核酸分子可包括shRNA、microRNA、siRNA和/或反义寡核苷酸。
进一步地,所述抑制SHCBP1表达的试剂包括shRNA,例如SEQ ID NO:1和2所示的shRNA。
发明的有益效果
本发明已发现SHCBP1的表达水平与抗HER2抗体治疗乳腺癌的效果相关,因而可以用于预测抗HER2抗体治疗乳腺癌的效果。
本发明还进一步发现抑制SHCBP1的表达水平可以提升抗HER2抗体治疗乳腺癌尤其是HER2低表达乳腺癌的效果。
附图说明
图1显示了MCF10A细胞、T47D细胞、ZR-75-1细胞和BT474细胞中HER2表达水平的验证以及在SHCBP1敲低后HER2的表达水平情况。
图2显示了SHCBP1在MCF10A细胞、T47D细胞、ZR-75-1细胞和BT474细胞中的表达水平以及在shRNA敲低后其表达水平的变化。
图3显示了在SHCBP1敲低前后,曲妥珠单抗和德卢替康-曲妥珠单抗在体外对T47D细胞、ZR-75-1细胞和BT474细胞的生长抑制情况。
图4显示了经曲妥珠单抗或德卢替康-曲妥珠单抗治疗后,T47D细胞、ZR-75-1细胞和BT474细胞中SHCBP1表达情况的变化。
图5显示了在SHCBP1敲低前后,曲妥珠单抗和德卢替康-曲妥珠单抗在体内对移植了ZR-75-1细胞或BT474细胞的荷瘤小鼠的肿瘤抑制情况。
图6显示了经曲妥珠单抗或德卢替康-曲妥珠单抗治疗后,荷瘤小鼠的肿瘤组织中SHCBP1表达水平的变化情况。
具体实施方式
以下结合具体实施例来进一步说明本发明,但实施例并不对本发明做任何形式的限定。除非特别说明,本发明采用的试剂、方法和设备为本技术领域常规试剂、方法和设备。
实施例1:SHCBP1在HER2低表达和HER2阳性乳腺癌细胞中高表达
本发明人先前已证实SHCBP1在乳腺癌细胞中的表达依赖于HER2的表达。与此一致地,分别采用MCF10A细胞(人正常乳腺上皮细胞)、T47D细胞(HER2阴性乳腺癌细胞)、ZR-75-1细胞(HER2低表达乳腺癌细胞)和BT474细胞(HER2阳性乳腺癌细胞),用含10%胎牛血清的RPMI-1640培养基(含100U·mL-1青霉素和0.1mg·mL-1链霉素),于37℃,5%CO2恒温培养箱中培养,细胞呈单层生长,达80%左右融合时收获细胞,提取RNA,利用RT-qPCR验证HER2表达量并检测SHCBP1表达水平。结果如图1和图2所示,其显示SHCBP1在BT474细胞和ZR-75-1细胞中的表达水平相较于MCF10A细胞和T47D细胞均升高。而当用shRNA(SEQ ID NO:1)敲低SHCBP1表达时,HER2表达量不受影响,表明SHCBP1在HER2下游发挥作用。
实施例2:SHCBP1在HER2低表达和HER2阳性乳腺癌细胞中的表达水平与治疗效果相关
分别采用T47D细胞、ZR-75-1细胞和BT474细胞,用含10%胎牛血清的RPMI-1640培养基(含100U·mL-1青霉素和0.1mg·mL-1链霉素),于37℃,5%CO2恒温培养箱中培养,细胞呈单层生长,达80%左右融合时传代。将对数生长期的各细胞用0.25%的胰酶和0.02%EDTA混合消化液消化收集后,进行细胞计数,用新鲜的RPMI-1640培养基稀释各细胞至2×104个/mL,500μL/孔接种于24孔板,置于37℃,5%CO2的饱和湿度培养箱中培养,培养24h后,更换培养液,进行给药,每种细胞均设置一式两份的空白对照组(无细胞)、阴性对照组(细胞培养液)、曲妥珠单抗治疗组(40μg/mL)和德卢替康-曲妥珠单抗(抗体-药物缀合物DS-8201)治疗组(40μg/mL)。
培养24h后,弃去各孔培养液,向一式两份中的一份的每孔加入20μL 5mg/mL的MTT溶液和80μL培养基,继续培养4h。弃去各孔残液,每孔加入二甲基亚砜100μL,置摇床上低速振荡30min,使结晶物充分溶解。用酶标仪检测490nm波长处测量各孔的吸光值(A490)。
细胞生长抑制率(%)=[(阴性对照组A490-空白组A490)-(给药组A490-空白组A490)]/(阴性对照组A490-空白组A490)×100%。
同时将一式两份中的另一份的每孔的细胞合并以提取RNA,利用RT-qPCR检测SHCBP1表达水平。
结果如图3所示,其显示曲妥珠单抗仅能够抑制BT474细胞的生长,而对T47D细胞和ZR-75-1细胞的生长没有抑制作用,与此相应地,如图4所示,经曲妥珠单抗治疗的BT474细胞中SHCBP1的表达水平降低,而ZR-75-1细胞中的SHCBP1表达水平则仍然较高;而德卢替康-曲妥珠单抗能够抑制BT474细胞和ZR-75-1细胞的生长(图3),而对T47D细胞的生长没有抑制作用,与此相应地,经德卢替康-曲妥珠单抗治疗的BT474细胞和ZR-75-1细胞中SHCBP1的表达水平降低(图4)。
根据本实施例的结果可知,SHCBP1的表达水平与抗HER2抗体的治疗效果相关,因而可用于预测抗HER2抗体治疗HER2阳性和HER2低表达乳腺癌的效果。
实施例3:SHCBP1抑制剂增强抗HER2抗体的治疗效果
构建靶向SHCBP1的shRNA序列(SEQ ID NO:1),连接于稳定表达质粒并进行病毒包装,病毒分别转染BT474细胞和ZR-75-1细胞后经过嘌呤霉素筛选和单克隆筛选,得到SHCBP1敲低的BT474细胞和ZR-75-1细胞。
如实施例2所述,分别用曲妥珠单抗和德卢替康-曲妥珠单抗处理SHCBP1敲低或未敲低的BT474细胞和ZR-75-1细胞,测试细胞生长抑制率。结果如图3所示,其显示敲低SHCBP1前后,曲妥珠单抗和德卢替康-曲妥珠单抗对BT474细胞的增殖抑制作用变化不大,然而敲低SHCBP1之后曲妥珠单抗对ZR-75-1细胞的增殖抑制作用显著增加,德卢替康-曲妥珠单抗对ZR-75-1细胞的增殖抑制作用也有一定增加,因此提示抑制SHCBP1的表达可以辅助增强抗HER2抗体对HER2低表达乳腺癌的治疗效果。
实施例4:SHCBP1的表达水平在体内动物模型中与抗HER2抗体治疗效果相关
取50只雌性裸鼠麻醉后,于裸鼠左前肢腋窝皮下注射200μL乳腺癌细胞株ZR-75-1细胞或BT474细胞,细胞数浓度为1×107/mL,接种后每两天观察裸鼠肿瘤生长情况及其全身状况,掌握肿瘤的形成时间及生长情况,观察小鼠一般活动及营养状态等。每两天用游标卡尺测量肿瘤长径,待肿瘤的直径长至5-7mm,即表明造模成功。筛选造模成功的裸鼠45只,随机分为3组,每组15只,分别为模型组、曲妥珠单抗组、德卢替康-曲妥珠单抗组。另取剩余15只正常裸鼠作为空白对照组,测定给药前各组裸鼠的体重。
各组给药剂量如下:
空白对照组:腹腔注射等体积的生理盐水;
模型组:腹腔注射等体积的生理盐水;
曲妥珠单抗组:腹腔注射50mg/kg的曲妥珠单抗;
德卢替康-曲妥珠单抗组:腹腔注射50mg/kg的德卢替康-曲妥珠单抗;
各组每周给药3次,连续给药4周。每天观察裸鼠的状态、如体重、食欲和精神等,4周后称重,处死裸鼠,取出肿瘤并称重,计算抑瘤率=(模型组瘤重-用药组瘤重)/模型组瘤重×100%,采用免疫组化方法检测肿瘤组织中SHCBP1的表达,并对裸鼠尸体进行全面尸检,肉眼观察裸鼠心、肝、肾、肺、脾、胸腺、肠道等主要器官的变化。
结果如图5和图6所示,其显示曲妥珠单抗的治疗对肿瘤的抑制作用有限(图5)且肿瘤组织中SHCBP1的表达较高(图6),而德卢替康-曲妥珠单抗的治疗对肿瘤有明显的抑制作用(图5)且肿瘤组织中SHCBP1的表达较低(图6)。这进一步证明了SHCBP1的表达水平与抗HER2抗体的治疗效果相关,因而可用于预测抗HER2抗体治疗HER2阳性和HER2低表达乳腺癌的效果。
实施例5:SHCBP1的抑制在体内动物模型中增强抗HER2抗体的治疗效果
如实施例4所述构建HER2低表达乳腺癌小鼠模型,不同之处在于注射SHCBP1敲低的ZR-75-1细胞或BT474细胞。分别用曲妥珠单抗和德卢替康-曲妥珠单抗对其进行治疗,计算抑瘤率。结果如图5所示,其显示经SHCBP1敲低后,曲妥珠单抗和德卢替康-曲妥珠单抗治疗的抑瘤率均有提升。这进一步证明抑制SHCBP1的表达可以显著增强抗HER2抗体对HER2低表达乳腺癌的治疗效果。
需要说明的是,本发明的说明书及其附图中给出了本发明的较佳的实施例,但是,本发明可以通过许多不同的形式来实现,并不限于本说明书所描述的实施例,这些实施例不作为对本发明内容的额外限制,提供这些实施例的目的是使对本发明的公开内容的理解更加透彻全面。并且,上述各技术特征继续相互组合,形成未在上面列举的各种实施例,均视为本发明说明书记载的范围;进一步地,对本领域普通技术人员来说,可以根据上述说明加以改进或变换,而所有这些改进和变换都应属于本发明所附权利要求的保护范围。
Claims (2)
1.抑制SHCBP1表达的试剂在制备用于增强抗HER2抗体治疗乳腺癌的效果的药物中的用途,其特征在于,所述乳腺癌是HER2低表达乳腺癌,所述抑制SHCBP1表达的试剂包括抑制SHCBP1表达的核酸分子,所述核酸分子包括shRNA,所述shRNA具有如SEQ ID NO:1所示的序列。
2.根据权利要求1所述的用途,其特征在于,所述抗HER2抗体选自曲妥珠单抗或德卢替康-曲妥珠单抗。
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Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110791566A (zh) * | 2019-10-29 | 2020-02-14 | 徐州市中心医院 | 人shcbp1基因的用途及相关产品 |
CN113785076A (zh) * | 2019-05-03 | 2021-12-10 | 株式会社递希真 | 预测癌症预后的方法及其组合物 |
CN114712512A (zh) * | 2021-01-06 | 2022-07-08 | 兰州大学第二医院 | 一种用于治疗her2阳性癌症的新型联合疗法 |
CN114736966A (zh) * | 2022-05-07 | 2022-07-12 | 中山大学肿瘤防治中心(中山大学附属肿瘤医院、中山大学肿瘤研究所) | 逆转乳腺癌耐药性的组合制剂及标志物应用 |
KR20230029059A (ko) * | 2021-08-23 | 2023-03-03 | 사회복지법인 삼성생명공익재단 | 트라스투주맙 저항성 예측용 신규 바이오마커 및 이의 용도 |
CN117100866A (zh) * | 2023-03-31 | 2023-11-24 | 兰州大学第二医院 | 一种抗肿瘤血管生成药物增效剂、抗肿瘤药物组合物及应用 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2008037700A2 (en) * | 2006-09-27 | 2008-04-03 | Siemens Healthcare Diagnostics Gmbh | Methods for breast cancer prognosis |
-
2024
- 2024-01-23 CN CN202410089744.9A patent/CN117607442B/zh active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113785076A (zh) * | 2019-05-03 | 2021-12-10 | 株式会社递希真 | 预测癌症预后的方法及其组合物 |
CN110791566A (zh) * | 2019-10-29 | 2020-02-14 | 徐州市中心医院 | 人shcbp1基因的用途及相关产品 |
CN114712512A (zh) * | 2021-01-06 | 2022-07-08 | 兰州大学第二医院 | 一种用于治疗her2阳性癌症的新型联合疗法 |
KR20230029059A (ko) * | 2021-08-23 | 2023-03-03 | 사회복지법인 삼성생명공익재단 | 트라스투주맙 저항성 예측용 신규 바이오마커 및 이의 용도 |
CN114736966A (zh) * | 2022-05-07 | 2022-07-12 | 中山大学肿瘤防治中心(中山大学附属肿瘤医院、中山大学肿瘤研究所) | 逆转乳腺癌耐药性的组合制剂及标志物应用 |
CN117100866A (zh) * | 2023-03-31 | 2023-11-24 | 兰州大学第二医院 | 一种抗肿瘤血管生成药物增效剂、抗肿瘤药物组合物及应用 |
Non-Patent Citations (5)
Title |
---|
A Novel 4-gene Score to Predict Survival, Distant Metastasis and Response to Neoadjuvant Therapy in Breast Cancer;Oshi M et al.;Cancers.;20200502;第12卷(第5期);全文 * |
Hyperactivation of HER2-SHCBP1-PLK1 axis promotes tumor cell mitosis and impairs trastuzumab sensitivity to gastric cancer;Wengui Shi;Nat Commun.;20210514;第12卷(第1期);全文 * |
SHCBP1 is over-expressed in breast cancer and is important in the proliferation and apoptosis of the human malignant breast cancer cell line;Wen Feng;Gene;20160801;第587卷(第1期);全文 * |
SHCBP1在非小细胞肺癌发生发展中的作用;罗小宁 等;现代医学;20210630;第49卷(第06期);全文 * |
乳腺癌CDK12伴随ERBB2基因共扩增对拉帕替尼治疗耐药的影响及机制研究;李慧;CNKI硕士电子期刊;20190215;第2019年卷(第02期);全文 * |
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