CN107429253B - Cs1靶向性嵌合抗原受体修饰的t细胞 - Google Patents
Cs1靶向性嵌合抗原受体修饰的t细胞 Download PDFInfo
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Abstract
描述了用于治疗恶性黑素瘤和其它表达CS1的癌症的嵌合抗原受体。
Description
对相关申请的交叉引用
本申请要求2014年12月5日提交的题为“USE OF CENTRAL MEMORY DERIVED-CS1CHIMERIC ANTIGEN RECEPTORMODIFIED T CELLS TO TREAT MULTIPLE MYELOMA”的美国非临时申请No.62/088,423的权益,其内容完整并入本文。
发明背景
已经研究了基于肿瘤特异性T细胞的免疫疗法,包括采用工程化T细胞的疗法用于抗肿瘤治疗。在某些情况下,此类疗法中使用的T细胞在体内未保持有活性达足够长的时段。在某些情况下,T细胞的抗肿瘤活性相对较低。因此,本领域中需要具有较长期的抗肿瘤功能的肿瘤特异性癌症治疗。
利用嵌合抗原受体(CAR)工程化T细胞的过继性T细胞疗法(ACT)可以提供安全且有效的方式来降低各种癌症的复发率,因为CART细胞可以工程化为以不依赖于MHC的方式特异性识别抗原性独特的肿瘤群体。
多发性骨髓瘤(MM)是以浆细胞克隆扩增为特征的B细胞恶性。MM占所有癌症的约1%,并且在美国略微超过10%的血液恶性。仅在美国,今年将诊断出约20,000例新病例,并且超过11,000人将死于此疾病。MM目前的疗法通常会诱导消退,但几乎所有的患者最终复发并死亡。
CS1是信号传导淋巴细胞活化分子(SLAM)受体家族的细胞表面糖蛋白,其在正常浆细胞和MM细胞上高度且选择性表达,在NK细胞上较低表达,并且在正常组织上很少表达或无表达。在复发性MM患者中与硼替佐米(bortezomib)一起给予的人源化CS1单克隆抗体Elotuzumabc(HuLuc63)在48%的患者中产生≥PR。MM细胞上的CS1高表达以及其限于正常组织中的浆细胞使CS1成为CAR T细胞疗法的合理靶物(Hsi et al.2008Clin Cancer Res14:2775)。
发明概述
本文描述了包含胞外域、跨膜域和胞内信号传导域的CAR。胞外域包含CS1特异性scFv区或其变体以及任选地间隔物,其包含例如人Fc域的部分。胞外域使得CAR在T细胞表面上表达时能够将T细胞活性引导至表达CS1(在MM表面上的受体)的细胞。跨膜域包含例如CD4跨膜域、CD8跨膜域、CD28跨膜域、或CD3跨膜域。胞内信号传导域包含来自人CD3复合物的zeta链(CD3ζ)的信号传导域和一个或多个共刺激域,例如4-1BB共刺激域。在胞内区中包含与CD3ζ串联的共刺激域,如4-1BB(CD137)共刺激域使得T细胞能够接受共刺激信号。T细胞,例如,患者特异性的自体T细胞可以工程化为表达本文所述的CAR,并且可以扩充工程化细胞并且在ACT中使用。可以使用各种T细胞亚组,包括αβT细胞和γδT细胞两者。此外,CAR可以在其它免疫细胞如NK细胞中表达。在用表达本文所述的CAR的免疫细胞治疗患者的情况下,细胞可以是自体T细胞或同种异体T细胞。在一些情况下,使用的细胞是包括CD4+和CD8+中央记忆T细胞(TCM)的细胞群体,其是CD62L+,CCR7+,CD45RO+和CD45RA-。细胞群体也可以包含其它类型的T细胞。
本文描述的CS1 CAR具有某些有益的特征,例如过继转移后的持久性和增强的抗肿瘤活性。
表达靶向CS1的CAR的T细胞可用于治疗癌症诸如MM以及表达CS1的其它癌症。因此,本公开内容包括使用表达本文所述的CAR的T细胞治疗表达CS1的癌症的方法。
本文描述了编码CAR的核酸分子,所述CAR包含CS1 scFv(例如,EVQLVESGGGLVQPGGSLRLSCAASGFDFSRYWMSWVRQAPGKGLEWIGEINPDSSTINYAPSLKDKFIISRDNAKNSLYLQMNSLRAEDTAVYYCARPDGNYWYFDVWGQGTLVTVSSGSTSGGGSGGGSGGGGSSDIQMTQSPSSLSASVGDRVTITCKASQDVGIAVAWYQQKPGKVPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQPEDVATYYCQQYSSYPYTFGQGTKVEIK;SEQ ID NO:1)或其具有1-5个(例如,1或2个)氨基酸修饰(例如,取代)的变体;选自以下的跨膜域:CD4跨膜域或其具有1-5个(例如,1或2个)氨基酸修饰(例如取代)的变体,CD8跨膜域或其具有1-5个(例如1或2个)氨基酸修饰(例如取代)的变体,CD28跨膜域或其具有1-5个(例如,1或2个)氨基酸修饰(例如取代)的变体和CD3ζ跨膜域或其具有1-5个(例如,1或2个)氨基酸修饰(例如,取代)的变体;共刺激域(例如,CD28共刺激域或其具有1-5个(例如,例如,1或2个)氨基酸修饰(例如取代)的变体;或4-1BB共刺激域或其具有1-5个(例如,1或2个)氨基酸修饰(例如取代)的变体;或CD28共刺激域或其具有1-5个(例如,1或2个)氨基酸修饰(例如,取代)的变体和4-1BB共刺激域或其具有1-5个(例如,1或2个)氨基酸修饰(例如,取代)的变体两者);和CD3ζ信号传导域或其具有1-5个(例如,1或2个)氨基酸修饰的变体。
本公开还涉及编码本文所述的任何CAR的核酸分子(例如,包含编码CAR之一的核酸序列的载体)和表达本文所述的任何CAR的分离的T细胞。
本文描述了编码嵌合抗原受体的核酸分子,其中嵌合抗原受体包含:CS1 scFv;间隔区;CD28或CD4跨膜域,CD28共刺激域或4-1BB共刺激域,任选的GlyGlyGly接头和CD3ζ信号传导域。
在一个实施方案中,CS1 CAR由SEQ ID NO:31、34、37、40、43和46(缺乏信号序列的成熟CAR)中任一项的氨基酸序列组成或包含SEQ ID NO:31、34、37、40、43和46(缺乏信号序列的成熟CAR)中任一项的氨基酸序列,或CS1 CAR由SEQ ID NO:30、33、36、39、42和45中任一项(具有GMCSFRa信号序列的未成熟CAR)的氨基酸序列组成或包含SEQ ID NO:30、33、36、39、42和45中任一项(具有GMCSFRa信号序列的未成熟CAR)的氨基酸序列。CAR可以以包含信号序列,例如人GM-CSF受体alpha信号序列(MLLLVTSLLLCELPHPAFLLIP;SEQ ID NO:26)的形式表达。CAR可以与另外的序列一起表达,所述另外的序列可用于监测表达,例如T2A跳跃(skip)序列和截短的EGFRt。因此,CAR可以包含SEQ ID NO:29-46中任一项的氨基酸序列或由SEQ ID NO:29-46中任一项的氨基酸序列组成,或者可以包含与SEQ ID NO:29-46中任一项至少95%,96%,97%,98%或99%相同的氨基酸序列或由与SEQ ID NO:29-46中任一项至少95%,96%,97%,98%或99%相同的氨基酸序列组成。CAR可以包含SEQ ID NO:29-46中任一项且具有多至1、2、3、4或5个氨基酸变化(优选保守氨基酸变化)的氨基酸序列或由SEQ ID NO:29-46中任一项且具有多至1、2、3、4或5个氨基酸变化(优选保守氨基酸变化)的氨基酸序列组成。
还公开了通过载体转导的人T细胞群体,所述载体包含编码本文所述的CS1嵌合抗原受体的表达盒(例如,包含SEQ ID NO:29-46中任一项的氨基酸序列或与SEQ ID NO:29-46中任一项至少95%,96%,97%,98%或99%相同的氨基酸序列或SEQ ID NO:29-46中任一项且具有多至1、2、3、4或5个氨基酸变化(优选保守氨基酸变化)的氨基酸序列或者由SEQID NO:29-46中任一项的氨基酸序列或与SEQ ID NO:29-46中任一项至少95%,96%,97%,98%或99%相同的氨基酸序列或SEQ ID NO:29-46中任一项且具有多至1、2、3、4或5个氨基酸变化(优选保守氨基酸变化)的氨基酸序列组成)。
在各种实施方案中:人T细胞群体是中央记忆T细胞(Tcm),例如CD8+/CD4+Tcm。
“氨基酸修饰”是指蛋白质或肽序列中的氨基酸取代、***和/或缺失。“氨基酸取代”或“取代”是指在亲本肽或蛋白质序列中的特定位置处的氨基酸用另一种氨基酸替换。可以进行取代从而以非保守的方式(即,通过将密码子从属于具有特定大小或特征的氨基酸分组的氨基酸改变为属于另一分组的氨基酸)或以保守的方式(即通过将密码子从属于具有特定大小或特征的氨基酸分组的氨基酸改变为属于相同分组的氨基酸)改变所得蛋白质中的氨基酸。此类保守变化通常导致所得蛋白质的结构和功能的变化较小。以下是氨基酸的各种分组的实例:1)具有非极性R基团的氨基酸:丙氨酸,缬氨酸,亮氨酸,异亮氨酸,脯氨酸,苯丙氨酸,色氨酸,甲硫氨酸;2)具有不带电荷的极性R基团的氨基酸:甘氨酸,丝氨酸,苏氨酸,半胱氨酸,酪氨酸,天冬酰胺,谷氨酰胺;3)具有带电荷的极性R基团的氨基酸(pH6.0时带负电荷):天冬氨酸,谷氨酸;4)碱性氨基酸(pH 6.0时带正电荷):赖氨酸,精氨酸,组氨酸(pH6.0)。另一分组可以是具有苯基基团的那些氨基酸:苯丙氨酸,色氨酸和酪氨酸。
CS1 scFv域
CS1ScFv域可以是结合CS1的任何ScFv。在一些情况下,CS1ScFv域包含与SEQ IDNO:1至少90%,至少95%,至少98%相同或与SEQ ID NO:1相同的序列。在一些情况下,与SEQ ID NO:1相比,CS1 scFv具有1、2、3、4或5个氨基酸变化(优选保守)(the CS1 scFv has1,2,3,4 of 5 amino acid changes(preferably conservative)compared to SEQ IDNO:1)。ScFv可以是人源化ScFv。
间隔区
本文描述的CAR可以包含位于CS1靶向域(即,aCS1 ScFv或其变体)和跨膜域之间的间隔区。可以使用多种不同的间隔物。它们中的一些包含人Fc区的至少部分,例如人Fc区的铰链部分或CH3域或其变体。下文表1提供了可用于本文所述的CAR的各种间隔物。
表1:间隔物的实例
一些间隔区包括整个或部分免疫球蛋白(例如,IgG1,IgG2,IgG3,IgG4)铰链区,即落在免疫球蛋白的CH1和CH2域之间的序列,例如IgG4Fc铰链或CD8铰链。一些间隔区包含免疫球蛋白CH3域或CH3域和CH2域两者。免疫球蛋白衍生的序列可以包含一个或多个氨基酸修饰,例如1、2、3、4或5个取代,例如减少脱靶结合的取代。
在某些实施方案中,间隔物是源自IgG1,IgG2,IgG3或IgG4的铰链/接头(linger),其包含用与未修饰铰链中存在的氨基酸残基不同的氨基酸残基取代的一个或多个氨基酸残基。一个或多个取代的氨基酸残基选自但不限于位置220,226,228,229,230,233,234,235,234,237,238,239,243,247,267,268,280,290,292,297,298,299,300,305,309,218,326,330,331,332,333,334,336,339或它们的组合处的一个或多个氨基酸残基。
在一些实施方案中,铰链/接头的经修饰的铰链源自IgG1,IgG2,IgG3或IgG4,其包括但不限于以下氨基酸残基取代中的一个或多个:C220S,C226S,S228P,C229S,P230S,E233P,V234A,L234V,L234F,L234A,L235A,L235E,G236A,G237A,P238S,S239D,F243L,P247I,S267E,H268Q,S280H,K290S,K290E,K290N,R292P,N297A,N297Q,S298A,S298G,S298D,S298V,T299A,Y300L,V305I,V309L,E318A,K326A,K326W,K326E,L328F,A330L,A330S,A331S,P331S,I332E,E333A,E333S,E333S,K334A,A339D,A339Q,P396L或它们的组合。
在一些实施方案中,经修饰的铰链源自具有SEQ ID NO:10或11的氨基酸序列或与SEQ ID NO:10或11至少90%,至少95%,至少98%相同的氨基酸序列的人IgG4铰链/CH2/CH3区。
在某些实施方案中,经修饰的铰链源自IgG4,其包含用与未修饰铰链中存在的氨基酸残基不同的氨基酸残基取代的一个或多个氨基酸残基。所述一个或多个取代的氨基酸残基选自但不限于位置220,226,228,229,230,233,234,235,234,237,238,239,243,247,267,268,280,290,292,297,298,299,300,305,309,218,326,330,331,332,333,334,336,339或它们的组合处的一个或多个氨基酸残基。
在一些实施方案中,经修饰的铰链源自IgG4,其包括但不限于以下氨基酸残基取代中的一个或多个:220S,226S,228P,229S,230S,233P,234A,234V,234F,234A,235A,235E,236A,237A,238S,239D,243L,247I,267E,268Q,280H,290S,290E,290N,292P,297A,297Q,298A,298G,298D,298V,299A,300L,305I,309L,318A,326A,326W,326E,328F,330L,330S,331S,331S,332E,333A,333S,333S,334A,339D,339Q,396L或其组合,其中未修饰铰链中的氨基酸在指定位置处用上文鉴定的氨基酸取代。在一个实例中,序列包括以下氨基酸变化S228P,L235E和N297Q。
对于本文中讨论的免疫球蛋白中的氨基酸位置,编号方式根据EU索引或EU编号方案(Kabat et al.1991 Sequences of Proteins of Immunological Interest,5th Ed.,United States Public Health Service,National Institutes of Health,Bethesda,在此通过引用并入)。EU索引或如Kabat中的EU索引或EU编号方案是指EU抗体的编号(Edelman等1969 Proc Natl Acad Sci USA 63:78-85)。
铰链/接头区还可以包含具有序列ESKYGPPCPSCP(SEQ ID NO:4)或ESKYGPPCPPCP(SEQ ID NO:3)的IgG4铰链区。
铰链/接头区还可以包含序列ESKYGPPCPPCP(SEQ ID NO:3),随后是接头序列GGGSSGGGSG(SEQ ID NO:2),随后是IgG4CH3序列GQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK(SEQID NO:12)。因此,整个接头/间隔区可包含序列:ESKYGPPCPPCPGGGSSGGGSGGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK(SEQ ID NO:11)。在一些情况下,与SEQ ID NO:11相比,间隔物具有1、2、3、4或5个单个氨基酸变化(例如保守变化)。在一些情况下,以减少Fc受体(FcR)的结合的方式在两个位置处突变的IgG4Fc铰链/接头区(L235E;N297Q)。
跨膜区
可以使用多种跨膜域。表2包括合适的跨膜域的实例。在存在间隔区的情况下,跨膜域位于间隔区的羧基末端。
表2:跨膜域的实例
共刺激域
共刺激域可以是适用于与CD3ζ信号传导域一起使用的任何域。在一些情况下,共刺激域是CD28共刺激域,其包含与以下序列至少90%,至少95%,至少98%相同或与以下序列相同的序列:RSKRSRHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS(SEQ ID NO:23;LL至GG氨基酸变化加双下划线)。在一些情况下,与SEQ ID NO:23相比,CD28共信号传导域具有1、2、3、4或5个氨基酸变化(1,2,3,4 of 5 amino acid)(优选保守,优选不在下划线的GG序列中)。在一些情况下,共信号传导域是4-1BB共信号传导域,其包含与以下的序列至少90%,至少95%,至少98%相同或与以下的序列相同的序列:KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL(SEQ ID NO:24)。在一些情况下,与SEQ ID NO:24相比,4-1BB共信号传导域具有1、2、3、4或5个氨基酸变化(优选保守的)。
共刺激域位于跨膜域和CD3ζ信号传导域之间。表3包括合适的共刺激域以及CD3ζ信号传导域序列的实例。
表3:CD3ζ域和共刺激域的实例
在各种实施方案中:共刺激域选自:表3中描述的共刺激域或其具有1-5个(例如1或2个)氨基酸修饰的变体,CD28共刺激域或其具有1-5个(例如,1或2个)氨基酸修饰的变体,4-1BB共刺激域或其具有1-5个(例如1或2个)氨基酸修饰的变体和OX40共刺激域或其具有1-5个(例如1或2个)氨基酸修饰的变体。在某些实施方案中,存在4-1BB共刺激域或其具有1-5个(例如,1或2个)氨基酸修饰的变体。在一些实施方案中,存在有两个共刺激域,例如CD28共刺激域或其具有1-5个(例如,1或2个)氨基酸修饰(例如取代)的变体和4-1BB共刺激域或其具有1-5个(例如,1或2个)氨基酸修饰(例如,取代)的变体。在各种实施方案中,1-5个(例如,1或2个)氨基酸修饰是取代。共刺激域在CD3ζ信号传导域的氨基末端,并且在一些情况下,由2-10,例如3个氨基酸(例如,GGG)组成的短接头位于共刺激域和CD3ζ信号传导域之间。
CD3ζ信号传导域
CD3ζ信号传导域可以是适用于与CD3ζ信号传导域一起使用的任何域。在一些情况下,CD3ζ信号传导域包含与以下序列至少90%,至少95%,至少98%相同或与以下序列相同的序列:RVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR(SEQ ID NO:21)。在一些情况下,与SEQID NO:21相比,CD3ζ信号传导具有1、2、3、4、或5个氨基酸变化(优选保守的)。
截短的EGFR
CD3ζ信号传导域之后可以是核糖体跳跃序列(例如,LEGGGEGRGSLLTCGDVEENPGPR;SEQ ID NO:27)和截短的EGFR,其具有与以下序列至少90%,至少95%,至少98%相同或与以下序列相同的序列:LVTSLLLCELPHPAFLLIPRKVCNGIGIGEFKDSLSINATNIKHFKNCTSISGDLHILPVAFRGDSFTHTPPLDPQELDILKTVKEITGFLLIQAWPENRTDLHAFENLEIIRGRTKQHGQFSLAVVSLNITSLGLRSLKEISDGDVIISGNKNLCYANTINWKKLFGTSGQKTKIISNRGENSCKATGQVCHALCSPEGCWGPEPRDCVSCRNVSRGRECVDKCNLLEGEPREFVENSECIQCHPECLPQAMNITCTGRGPDNCIQCAHYIDGPHCVKTCPAGVMGENNTLVWKYADAGHVCHLCHPNCTYGCTGPGLEGCPTNGPKIPSIATGMVGALLLLLVVALGIGLFM(SEQ IDNO:28)。在一些情况下,与SEQ ID NO:28相比,截短的EGFR具有1、2、3、4或5个氨基酸变化(优选保守的)。
CS1 CAR
CS1 CAR可以包含与图2,图6,图7,图8,图9或图10中描绘的氨基酸序列(SEQ IDNO:29-46;包括或排除GMCSFRa信号序列并且包括或排除T2A核糖体跳跃序列和截短的EGFRt)至少90%,至少95%,至少98%相同或相同的序列。
在本文描述的靶向CS1的CAR中的是表4中汇总的那些,其中指示每个CAR的间隔区,跨膜域和共刺激域。
表4:靶向CS1的CAR的实例
*SEQ ID NO代表:包含GMCSFRa信号序列,T2A和EGFRt的描绘的整个序列//包含GMCSFRa信号序列但是排除T2A和EGFRt的序列//排除GMCSFRa信号序列,T2A和EGFRt的序列的序列。
附图简述
图1是表达慢病毒载体(CS1scFv-IgG4(HL-CH3)-CD28gg-Zeta(CO)-T2A-EGFRt_epHIV7)的CS1 CAR的示意图。CS1 CAR构建体包含:GMCSF信号序列,CS1 scFv,IgG4铰链区,接头,CH3域,CD28共刺激域和CD3ζ信号传导域。CAR构建体之后是T2A核糖体跳跃序列,然后是***基因EGFRt编码序列。CAR和EGFRt分子由单一转录物表达。
图2描绘了CS1 CAR的氨基酸序列,其包括信号肽,核糖体跳跃序列和EGFRt(SEQID NO:29)。
图3是描述研究结果的一对图,其显示CS1 CAR重新导向的Tcm(CS1 CAR re-directed Tcm)表现出针对MM细胞的细胞毒性。在与51Cr标记的靶细胞共培养后,使用4小时51Cr释放测定法评估增殖的CS1 CAR T细胞的细胞毒性。使用表达OKT3的LCL作为阳性对照,因为它们结合所有TCR,并且CS1阴性AML细胞(KG1a)用作阴性对照。CS1 CAR,而非未工程化的模拟(mock)T细胞显示针对MM细胞的特异性细胞毒性。
图4描述了研究的结果,显示CS1 CAR重新导向的Tcm细胞响应于MM细胞的刺激表现出效应器功能。在96孔组织培养板中将CS1 CAR T细胞(105)与作为刺激剂的105个MM.1S细胞共培养6小时。用流式细胞术分析107a脱粒和胞内IFNγ产生。在与MM细胞结合后诱导由爱必妥(Erbitux)鉴定的大多数CAR T细胞脱粒,并且响应抗原刺激检测到IFNγ阳性细胞。
图5描绘了研究的结果,其显示CS1 CAR重新导向的Tcm细胞在体内消除多发性骨髓瘤。通过胫骨内注射将约2x106个表达萤火虫萤光素酶的MM.1S细胞接种到NSG小鼠中。肿瘤接种后7天,通过静脉内注射将1x106个CS-1 CAR T细胞输注到携带肿瘤的小鼠中。一周一次用成像监测肿瘤负荷。使用接受未工程化细胞的小鼠作为对照。CS1 CART细胞在T细胞输注后14天完全消除MM肿瘤,而未工程化的T细胞对肿瘤抑制没有影响。
图6描绘了CS1scFv-IgG4(HL-CH3)-CD4tm-41BB-Zeta-T2A-EGFRt的氨基酸序列(SEQ ID NO:32)。
图7描述了CS1scFv-IgG4(L235E,N297Q)-CD4tm-41BB-Zeta-T2A-EGFRt的氨基酸序列(SEQ ID NO:35)。
图8描述了CS1scFv-IgG4(L235E,N297Q)-CD28tm-CD28gg-Zeta-T2A-EGFRt的氨基酸序列(SEQ ID NO:38)。
图9描述了CS1scFv-Linker-CD4tm-41BB-Zeta-T2A-EGFRt的氨基酸序列(SEQ IDNO:41)。
图10描述了CS1scFv-Linker-CD28tm-CD28gg-Zeta-T2A-EGFRt的氨基酸序列(SEQID NO:44)。
图11是CS1scFv-IgG4(HL-CH3)-CD28gg-Zeta-T2A-EGFRt_epHIV7的完整核苷酸序列(SEQ ID NO:47)。
图12描述了研究结果,其显示CS1 CAR重新导向的Tcm细胞在体内消除多发性骨髓瘤。在第-7天通过胫骨内注射将2x106个GFPffluc+MM.1S个细胞接种到NSG小鼠中。将1x106个中央记忆T细胞(Tcm)衍生的CS1 CAR+T细胞在第0天静脉内输注入携带肿瘤的小鼠中。未接受来自相同供体的T细胞或未转导的Tcm的小鼠用作阴性对照。通过生物光子成像(biophotonic imaging)监测肿瘤信号。描绘了来自多只小鼠的光子/秒的平均值±SEM。CAR是图2(CH2CD28);图6(CH2 4IBB);图8(EQ CD28);图7(EQ 4IBB);图10(L CD28)和图9(LCD4 IBB)的CAR。
发明详述
下文描述了几种CS1特异性嵌合抗原受体(“CAR”)的结构、构建和表征。CAR是包含胞外识别域、跨膜区和胞内信号传导域的重组生物分子。因此,术语“抗原”不限于结合抗体的分子,而是可以特异性结合任何受体的任何分子。因此,“抗原”是指CAR的识别域。胞外识别域(也称为胞外域或仅仅通过其含有的识别元件提及)包含特异性结合存在于靶细胞的细胞表面上的分子的识别元件。跨膜区将CAR锚定在膜中。胞内信号传导域包含来自人CD3复合物的zeta链的信号传导域,并任选地包含一个或多个共刺激信号传导域。CAR既可以结合抗原又能转导T细胞活化,不依赖于MHC限制。因此,CAR是“通用”免疫受体,其可以治疗具有抗原阳性肿瘤的患者群体,而不管其HLA基因型如何。使用表达肿瘤特异性CAR的T淋巴细胞的过继免疫疗法可以是治疗癌症的有力治疗策略。
在一些情况下,可以使用载体产生CS1 CAR,其中CAR可读框之后是T2A核糖体跳跃序列和截短的EGFR(EGFRt),其缺少细胞质信号传导尾部。在此排列中,EGFRt的共表达提供了惰性的、非免疫原性的表面标志物,其允许准确测量基因修饰的细胞,并且实现基因修饰的细胞的阳性选择,以及过继转移后治疗性T细胞的体内有效细胞追踪。有效控制增殖以避免细胞因子风暴和脱靶毒性是T细胞免疫疗法成功的重要障碍。在CS1CAR慢病毒载体中掺入的EGFRt可以起***基因作用以在治疗相关毒性的情况下消融CAR+T细胞。
本文描述的CAR可以通过本领域已知的任何手段产生,尽管优选使用重组DNA技术产生它。编码嵌合受体的几个区域的核酸可通过本领域已知的分子克隆的标准技术(基因组文库筛选,重叠PCR,引物辅助连接,定点诱变等)(其是常规的)制备并组装成完整的编码序列。优选地,将所得的编码区***表达载体中并用于转化合适的表达宿主细胞系,优选T淋巴细胞细胞系,最优选自体T淋巴细胞细胞系。
可以用用于CAR表达的载体转导从患者分离的各种T细胞亚群。中央记忆T细胞是一个有用的T细胞亚组。通过选择CD45RO+/CD62L+细胞,可以使用例如免疫磁性选择表达期望受体的细胞,从外周血单核细胞(PBMC)分离中央记忆T细胞。针对中央记忆T细胞富集的细胞可用抗CD3/CD28活化,用例如慢病毒载体转导,所述慢病毒载体指导表达CS1 CAR以及非免疫原性表面标志物,用于体内检测,消融,和潜在的离体选择。可以用IL-2/IL-15体外扩增活化/遗传修饰的CS1中央记忆T细胞,然后冷冻保存。
实施例1:用于表达CS1特异性CAR的epHIV7的构建和结构
pHIV7质粒是可以衍生表达CS1 CAR的临床载体的亲本质粒。用于表达CAR的epHIV7载体由pHIV7载体产生(Wang et al.2011 Blood 118:1255)。重要的是,该载体使用人EF1启动子来驱动CAR的表达。载体的5’和3’序列两者均源自pv653RSN,如先前源自HXBc2前病毒。多聚嘌呤段DNA瓣序列(cPPT)源自来自NIH AIDS试剂库(NIH AIDSReagentRepository)的HIV-1株pNL4-3。
如下进行pHIV7的构建。简言之,如下将含有来自gag-pol的653bp加5’和3’长末端重复(LTR)及居间SL3-新霉素磷酸转移酶基因(Neo)的pv653RSN亚克隆到pBluescript中:在步骤1中,从5’LTR到转录响应元件(RRE)的序列制成p5’HIV-151,然后通过除去TATA盒上游的序列修饰5’LTR,并先连接到CMV增强子,然后连接到SV40复制起点(p5'HIV-2)。在步骤2中,将3'LTR克隆到pBluescript中以制备p3’HIV-1后,在3'LTR增强子/启动子中进行400bp的缺失以除去HIV U3中的顺式调节元件并形成p3'HIV-2。在步骤3中,将从p5'HIV-3和p3'HIV-2分离的片段连接以制备pHIV-3。在步骤4中,通过除去额外的上游HIV序列进一步修饰p3'HIV-2以产生p3’HIV-3,并将含有WPRE的600bp BamHI-SalI片段添加到p3’HIV-3以制备p3'HIV-4。在步骤5中,通过PCR在大小中降低pHIV-3RRE,并连接到来自pHIV-3(未显示)的5’片段并且连接到p3’HIV-4,以制备pHIV-6。在步骤6中,从pNL4-3扩增含有来自HIV-1pNL4-3(55)的cPPT DNA瓣序列的190bp的BglII-BamHI片段,并将其置于pHV6中的RRE和WPRE序列之间以制备pHIV-7。使用此亲本质粒pHIV7-GFP(GFP,绿色荧光蛋白)用四质粒***包装亲本载体。
包装信号PSIψ是将病毒基因组有效包装到载体中需要的。RRE和WPRE增强RNA转录物转运和转基因的表达。已经证实了与WPRE组合的瓣序列(flap sequence)增强慢病毒载体在哺乳动物细胞中的转导效率。
对于产生病毒载体所需要的辅助功能分为三种不同的质粒,以减少通过重组产生有复制能力的慢病毒的可能性:1)pCgp编码病毒载体组装所需要的gag/pol蛋白;2)pCMV-Rev2编码Rev蛋白,其作用于RRE序列以帮助病毒基因组的运输,用于有效的包装;和3)pCMV-G编码水疱-口炎病毒(VSV)的糖蛋白,其是病毒载体的感染性所需要的。
在pHIV7编码的载体基因组和辅助质粒之间存在最小的DNA序列同源性。同源性区包括位于pCgp辅助质粒的gag/pol序列中的约600个核苷酸的包装信号区;所有三个辅助质粒中的CMV启动子序列;和辅助质粒pCgp中的RRE序列。高度不可能的是,有复制能力的重组病毒可以由于这些区域中的同源性而产生,因为它将需要多个重组事件。另外,任何得到的重组体将缺少慢病毒复制所需要的功能性LTR和tat序列。
通过EF1α-HTLV启动子(EF1p)替换CMV启动子,并且新质粒命名为epHIV7。EF1p具有563bp,并且在切除CMV启动子后,使用NruI和NheI引入epHIV7中。
已经从该***中除去了慢病毒基因组,其排除对于野生型病毒的致病性所必需,并且是靶细胞的生产性感染所需要的gag/pol和rev。此外,epHIV7载体构建体不含完整的3’LTR启动子,因此靶向细胞中得到的表达和逆转录的DNA原病毒基因组将具有无活性的LTR。由于此设计,无HIV-1衍生的序列将从原病毒转录,并且仅治疗序列将从它们各自的启动子表达。预期SIN载体中LTR启动子活性的除去显著降低宿主基因的无意激活的可能性。表5汇总了epHIV7中存在的各种调节物元件。
图1是CS1 CAR(CS1scFv-IgG4(HL-CH3)-CD28gg-Zeta(CO)-T2A-EGFR t_epHIV7),含有由CS1 scFv,IgG4铰链区,接头,CD28共刺激域和CD3ζ信号传导域构成的CAR构建体的慢病毒载体的示意图。CAR构建体之后是T2A核糖体跳跃序列,并且然后是***基因EGFRt编码序列。CAR和EGFRt分子由单一转录物表达。载体的全部核苷酸序列示于图11中,并且表5呈现了载体的各种元件的位置。
实施例2:用于转导患者T细胞的载体的产生
对于每个质粒(CS1 CAR_epHIV7;pCgp;pCMV-G;和pCMV-Rev2),产生种子库,其用于接种发酵罐以产生足够量的质粒DNA。对质粒DNA测试身份、不育性(sterility)和内毒素,之后其在产生慢病毒载体中使用。
简言之,从293T工作细胞(WCB)扩增细胞,所述293T工作细胞已经经过测试以确认不育性和病毒污染的缺乏。将来自293T WCB的293T细胞小瓶解冻。培养细胞并且扩增,直到存在足够数量的细胞以铺板合适数目的10层细胞工厂(CF)用于载体生产和细胞训练(celltrain)维持。细胞的单训练可用于生产。
以多至10CF的分批次生产慢病毒载体。可以在同一周生产两个分批次,导致大约20L慢病毒上清液/周的生产。在下游处理阶段期间,将所有分批次生产的材料合并,以便产生一批产物。将293T细胞铺板于293T培养基(含有10%FBS的DMEM)中的CF中。将工厂放置在37℃培养器中并水平调平,以获得CF的所有层上的细胞的均匀分布。两天后,使用CaPO4方法用上述四种慢病毒质粒转染细胞,所述CaPO4方法牵涉Tris:EDTA、2M CaCl2、2XHBS和四种DNA质粒的混合物。转染后第3天,收集含有分泌性慢病毒载体的上清液,纯化并浓缩。从CF中除去上清液后,从每个CF中收集生产终期细胞(End-of-Production Cell)。将细胞从每个工厂进行胰蛋白酶处理并通过离心收集。将细胞重悬于冷冻培养基中并冷冻保存。随后,将这些细胞用于有复制能力的慢病毒(RCL)检测。
为了纯化和配制载体,通过膜过滤澄清粗制上清液,以除去细胞碎片。通过内切核酸酶消化降解宿主细胞DNA和残留质粒DNA。病毒上清液用0.45μm滤器澄清细胞碎片。将澄清的上清液收集到添加有(终浓度50U/mL)的预先称重的容器中。残留质粒DNA和宿主基因组DNA的核酸内切酶消化在37℃下进行6小时。使用内切核酸酶处理的上清液的初始切向流超滤(TFF)浓度从粗制上清液中除去残留的低分子量组分,同时浓缩病毒约20倍。将澄清的内切核酸酶处理的病毒上清液通过具有500kD的NMWCO的中空纤维筒以设计为将剪切速率维持于约4,000sec-1或更小的流速循环,同时最大化流量速率。在浓缩过程期间启动核酸酶处理的上清液的渗滤以保持筒性能。使用PBS中的4%乳糖作为渗滤缓冲液,建立了80%渗透物置换率。病毒上清液达到目标体积,代表粗制上清液的20倍浓度,并且渗滤继续再进行4个交换体积,渗透物替换率为100%。
通过使用高速离心技术实现病毒产物的进一步浓缩。使用Sorvall RC-26 plus离心机以6000RPM(6,088RCF)在6℃将每个分批次的慢病毒沉淀16-20小时。然后,将来自每个分批的病毒团粒用PBS中的4%乳糖在50mL体积中重建。该缓冲液中重建的团粒代表病毒制备的最终制剂。整个载体浓缩过程导致大约200倍体积降低。在完成所有分批次之后,然后将材料置于-80℃,期间对来自每个分批次的样品测试不育性。在确认样品不育性后,在频繁搅拌的情况下在37℃迅速解冻分批次。然后,将材料合并并在病毒区套间中在II类A/B3生物安全柜中手动等分取样。使用在无菌USP 6类,外螺纹O-环冷冻小瓶中1mL浓缩慢病毒的填充构造。在COH的应用技术开发中心(Center for Applied Technology Development,CATD)的质量体系(Quality Systems,QS)根据CBG的政策和标准操作程序(Policies andStandard Operating Procedures for the CBG)并且符合现行的“良好制造实践”(GoodManufacturing Practices cGMPs),发布了所有材料。
为了确保慢病毒载体制剂的纯度,对它测试残留的宿主DNA污染物,以及残留宿主和质粒DNA的转移。在其它测试中,通过RT-PCR评估载体身份,以确保存在正确的载体。意图用于在本研究中使用的载体满足所有释放标准。
实施例3:适合于在ACT中使用的Tcm细胞的制备
通过白细胞单采术(leukopheresis)从患者获得T淋巴细胞,并且将适当的同种异体或自体T细胞亚组(例如中央记忆T细胞(Tcm))遗传改变以表达CAR,然后通过任何临床可接受的手段施用回患者,以实现抗癌疗法。
基本上如Wang等人(J Immunology 35:689,2012)中描述的那样分离CD8+的Tcm。简言之,在白细胞单采术当天,通过经Ficoll-Paque的密度梯度离心分离PBMC,随后在PBS/EDTA中清洗2次。然后,将PBMC在PBS中清洗一次,重悬于含有10%胎牛血清(FCS)的XVivo15培养基中,转移到300cc转移袋中,并在室温(RT)下储存在3-D旋转器上过夜。次日,在RT在含有10%FCS的X Vivo15中在具有临床级抗CD4(2.5mL)、抗CD14(1.25mL)、和抗CD45RA(2.5mL)微珠(Miltenyi Biotec)的300cc转移袋中将多至5x109个PBMC温育30分钟。然后根据制造商的说明书(Miltenyi Biotec),使用CliniMACSTM耗尽模式立即耗尽CD4+,CD14+和CD45RA+细胞。离心后,将细胞的未标记的阴性级份重悬于含有0.5%人血清白蛋白(HSA)的CliniMACSTMPBS/EDTA缓冲液(Miltenyi Biotec)中,然后在RT以0.1mg/106个细胞用临床级生物素化DREG56mAb(COHNMC CBG)标记30分钟。然后将细胞洗涤并重悬于含有0.5%HSA的100mL CliniMACSTM PBS/EDTA的终体积中,并转移到新的300cc转移袋中。与1.25mL抗生物素微珠(Miltenyi Biotec)温育30分钟后,根据制造商的说明书,用CliniMACSTM进行阳性选择纯化PBMC(CD8+TCM)的CD62L+级分,并重悬于含有10%FCS的XVivo15中。
通过使用前述方法的修改(modification),将CD4+,CD14+和CD45RA+选择修改为CD14+和CD45RA+选择,来制备CD8+/CD4+的Tcm。方法在CliniMACSTM设备上使用两步法首先消耗CD14+和CD45RA+细胞,然后阳性选择CD62L+细胞。该修改的平台从单一白细胞单采术产生50x106体积Tcm。
富集后,在完全X-Vivo15plus 50IU/mL IL-2和0.5ng/mL IL-15中配制Tcm细胞,并且转移到Teflon细胞培养袋,在那里用Dynal ClinExTMVivo CD3/CD28珠刺激它们。刺激后多至5天,以约3的感染复数(MOI)用编码CS1 CAR的慢病毒载体转导细胞。将培养物维持长达42天,添加完全X-Vivo15和IL-2和IL-15细胞因子,如细胞扩增需要的(保持细胞密度在3x105和2x106个活细胞/mL之间,并且培养的每周一、周三和周五补充细胞因子)。在21天内在这些条件下细胞通常扩增至约109个细胞。培养期结束时,收获细胞,洗涤两次,并且在临床级冷冻保存培养基中配制。
在T细胞输注当天,将冷冻保存且释放的产物解冻,清洗,并配制用于再输注。从液氮贮存中取出含有释放的细胞产物的冷冻保存的小瓶,解冻,冷却,并用PBS/2%人血清白蛋白(HSA)清洗缓冲液清洗。离心后,除去上清液,并且将细胞重悬于无防腐正常盐水(PFNS)/2%HSA输注稀释液中。除去样品以进行质量控制测试。
实施例4:CS1 CAR(CS1scFv-IgG4(HL-CH3)-CD28tm-CD28gg-Zeta-T2
A-EGFRt)的
氨基酸序列
图2中描绘了CS1scFv-IgG4(HL-CH3)-CD28tm-CD28gg-Zeta-T2A-EGF Rt的完整的氨基酸序列。整个序列(SEQ ID NO:29)。包含:22个氨基酸的GMCSF信号肽(SEQ ID NO:26)、CS1 scFv序列(SEQ ID NO:1);IgG4铰链序列(SEQ ID NO:3;氨基酸取代S至P有阴影);10个氨基酸的接头(SEQ ID NO:2);IgG4CH3序列(SEQ ID NO:12);28个氨基酸的CD28跨膜域序列(SEQ ID NO:14);CD28gg共刺激域序列(SEQ ID NO:23;LL至GG氨基酸变化突出显示);3个氨基酸的Gly接头;112个氨基酸的CD3ζ序列(SEQ ID NO:21);24个氨基酸的T2A跳跃序列(SEQ ID NO:27);和EGFRt序列(SEQ ID NO:28)。
实施例5:CS1 CAR的活性
在与51Cr标记的MM细胞(MM.1S)共培养后,使用4小时51Cr释放测定法评估表达图2所示的CAR的增殖CS1 CAR T细胞的细胞毒性。如图3中所示,工程化CS1 CAR T细胞表现出对MM细胞的特异性且有效的杀伤,而未转导的模拟T细胞对MM细胞没有细胞毒性。当与MM细胞共培养时,工程化的CS1 CAR Tcm介导的强效应器功能如107a脱粒和IFNgamma指示,如图4中所示。在过继性转移到携带MM肿瘤的NSG小鼠中时,CS1特异性T细胞表现出有效的抗肿瘤活性,如图5中所示。
在另一项用另外的CS1 CAR的研究中(图2和图6-10),在第7天通过胫骨内注射将2x106个GFPffluc+MM.1S细胞接种入NSG小鼠中。将1x106个中央记忆T细胞(Tcm)衍生的CS1CAR+T细胞在第0天静脉内输注入携带肿瘤的小鼠中。未接受来自相同供体的T细胞或未转导的Tcm的小鼠用作阴性对照。通过生物光子成像监测肿瘤信号。描绘了来自多只小鼠的光子/秒的平均值±SEM。图12中显示了此分析的结果。
Claims (29)
1.核酸分子,其编码包含SEQ ID NO:29、30、31、32、33、34、38、39和40中任一个的氨基酸序列的多肽。
2.权利要求1的核酸分子,其中所述多肽包含SEQ ID NO:29、30和31中任一个的氨基酸序列。
3.权利要求1的核酸分子,其中所述多肽包含SEQ ID NO:32、33和34中任一个的氨基酸序列。
4.权利要求1的核酸分子,其中所述多肽包含SEQ ID NO:38、39和40中任一个的氨基酸序列。
5.由包含编码多肽的表达盒的载体转导的人T细胞群体,所述多肽包含SEQ ID NO:29、30、31、32、33、34、38、39和40中任一个的氨基酸序列。
6.权利要求5的人T细胞群体,其中至少20%的经转导的人T细胞是中央记忆T细胞。
7.权利要求5的人T细胞群体,其中至少30%的经转导的人T细胞是中央记忆T细胞。
8.权利要求5的人T细胞群体,其中至少40%的经转导的人T细胞是中央记忆T细胞。
9.权利要求5的人T细胞群体,其中至少50%的经转导的人T细胞是中央记忆T细胞。
10.权利要求5的人T细胞群体,其中至少60%的经转导的人T细胞是中央记忆T细胞。
11.权利要求5的人T细胞群体,其中至少70%的经转导的人T细胞是中央记忆T细胞。
12.权利要求5的人T细胞群体,其中至少80%的经转导的人T细胞是中央记忆T细胞。
13.权利要求5的人T细胞群体,其中至少10%的经转导的中央记忆T细胞是CD4+。
14.权利要求5的人T细胞群体,其中至少20%的经转导的中央记忆T细胞是CD4+。
15.权利要求5的人T细胞群体,其中至少10%的经转导的中央记忆T细胞是CD8+。
16.权利要求5的人T细胞群体,其中至少20%的经转导的中央记忆T细胞是CD8+。
17.权利要求5的人T细胞群体,其中至少10%的中央记忆T细胞是CD4+并且至少10%是CD8+。
18.权利要求5的人T细胞群体,其中所述多肽包含SEQ ID NO:29、30和31中任一个的氨基酸序列。
19.权利要求5的人T细胞群体,其中所述多肽包含SEQ ID NO:32、33和34中任一个的氨基酸序列。
20.权利要求5的人T细胞群体,其中所述多肽包含SEQ ID NO:38、39和40中任一个的氨基酸序列。
21.权利要求5的人T细胞群体,其中所述多肽由SEQ ID NOs: 31、34和40中的任一个氨基酸序列组成。
22.在治疗表达CS-1的癌症的方法中使用的组合物,所述方法包括对有此需要的患者施用包含权利要求5-21中任一项的人T细胞群体的药物组合物。
23.权利要求22中使用的组合物,其中所述人T细胞群体对于所述患者是自体的。
24.权利要求22中使用的组合物,其中所述人T细胞群体对于所述患者是同种异体的。
25.权利要求22-24中任一项使用的组合物,其中所述癌症是多发性骨髓瘤。
26.包含权利要求5-21中任一项的人T细胞群体的药物组合物在制备组合物中的用途,所述组合物用于对有此需要的患者治疗表达CS-1的癌症。
27.权利要求26的用途,其中所述人T细胞群体对于所述患者是自体的。
28.权利要求26的用途,其中所述人T细胞群体对于所述患者是同种异体的。
29.权利要求26-28中任一项的用途,其中所述癌症是多发性骨髓瘤。
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Families Citing this family (40)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107429253B (zh) | 2014-12-05 | 2021-11-05 | 希望之城公司 | Cs1靶向性嵌合抗原受体修饰的t细胞 |
BR112017015453A2 (pt) | 2015-01-26 | 2018-01-23 | Cellectis | células imunes modificadas knockout para receptor de células t, dotadas de receptores quiméricos de antígeno, de ligação a cd123 para o tratamento de linfoma mieloide agudo recorrente/refratário ou neoplasia blástica de células dendríticas plasmacitoides |
EP3408297A2 (en) | 2016-01-29 | 2018-12-05 | Med Manor Organics, (P) Ltd | A chimeric antigen receptor specific to b-cell maturation antigen, a recombinant expression vector and a method thereof |
CU20180120A7 (es) | 2016-04-01 | 2019-05-03 | Kite Pharma Inc | Moléculas de unión a bcma |
US10603380B2 (en) | 2016-04-01 | 2020-03-31 | Kite Pharma, Inc. | Chimeric antigen and T cell receptors and methods of use |
HUE060645T2 (hu) | 2016-04-01 | 2023-04-28 | Kite Pharma Inc | Kiméra receptorok és felhasználásuk módszerei |
EP3478722A1 (en) | 2016-06-30 | 2019-05-08 | F. Hoffmann-La Roche AG | Improved adoptive t-cell therapy |
CN105949324B (zh) * | 2016-06-30 | 2019-08-27 | 上海恒润达生生物科技有限公司 | 靶向gpc3的嵌合抗原受体及其用途 |
US20190263928A1 (en) * | 2016-09-30 | 2019-08-29 | Baylor College Of Medicine | Adaptive chimeric antigen receptor t-cell design |
WO2018156711A1 (en) * | 2017-02-22 | 2018-08-30 | H. Lee Moffitt Cancer Center And Research Institute, Inc. | Il13ra2-binding chimeric antigen receptors |
EP3589295A4 (en) | 2017-02-28 | 2020-11-04 | Endocyte, Inc. | COMPOSITIONS AND METHODS OF T CAR LYMPHOCYTE THERAPY |
WO2018169922A2 (en) | 2017-03-13 | 2018-09-20 | Kite Pharma, Inc. | Chimeric antigen receptors for melanoma and uses thereof |
BR112019019606A2 (pt) * | 2017-03-20 | 2020-04-14 | Hope City | células t modificadas pelo receptor de antígeno quimérico direcionado a cs1 |
SG11201908796XA (en) | 2017-03-27 | 2019-10-30 | Hoffmann La Roche | Improved antigen binding receptors |
JOP20180042A1 (ar) | 2017-04-24 | 2019-01-30 | Kite Pharma Inc | نطاقات ربط مولد ضد متوافقة مع البشر وطرق الاستخدام |
US11149073B2 (en) | 2017-04-28 | 2021-10-19 | Julius-Maximilians-Universität Würzburg | ROR1-specific chimeric antigen receptors (CAR) with humanized targeting domains |
CN111542323A (zh) * | 2017-08-01 | 2020-08-14 | 尤利乌斯·马克西米利安维尔茨堡大学 | Flt3 car-t细胞和flt3抑制剂在治疗急性髓细胞性白血病中的用途 |
IL298699B2 (en) | 2017-08-11 | 2024-01-01 | Idac Holdings Inc | Traffic routing and switching between multiple access networks |
BR112020008340A2 (pt) * | 2017-11-01 | 2020-11-17 | Juno Therapeutics Inc | processo para gerar composições terapêuticas de células modificadas |
CN111727250A (zh) * | 2017-11-14 | 2020-09-29 | 阿奇利克斯股份有限公司 | 含有d结构域的多肽及其用途 |
US11464803B2 (en) | 2017-11-14 | 2022-10-11 | Arcellx, Inc. | D-domain containing polypeptides and uses thereof |
WO2019099707A1 (en) | 2017-11-16 | 2019-05-23 | Kite Pharma, Inc | Modified chimeric antigen receptors and methods of use |
WO2019144095A1 (en) | 2018-01-22 | 2019-07-25 | Seattle Children's Hospital (dba Seattle Children's Research Institute) | Methods of use for car t cells |
US20210040449A1 (en) | 2018-02-16 | 2021-02-11 | Kite Pharma, Inc. | Modified pluripotent stem cells and methods of making and use |
WO2019222796A1 (en) * | 2018-05-21 | 2019-11-28 | Carina Biotech Pty Ltd | Chimeric antigen receptors with modified linker domains and uses thereof |
US11951131B2 (en) | 2018-07-03 | 2024-04-09 | The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Anti-SLAMF7 chimeric antigen receptors |
US20210308184A1 (en) | 2018-08-01 | 2021-10-07 | City Of Hope | Tag72 targeted chimeric antigen receptor modified t cells for treatment of tag72-positive tumors |
CN112469734A (zh) * | 2018-08-03 | 2021-03-09 | 南京驯鹿医疗技术有限公司 | 表达嵌合抗原受体的t细胞、该嵌合抗原相关表达载体以及它们的应用 |
MX2021006968A (es) | 2018-12-12 | 2021-10-13 | Kite Pharma Inc | Antígeno quimérico y receptores de las células t, y métodos de uso. |
KR20210148293A (ko) | 2019-04-03 | 2021-12-07 | 프리시젼 바이오사이언시스 인코포레이티드 | 마이크로RNA-적응 shRNA(shRNAmiR)를 포함하는 유전자-변형 면역 세포 |
CA3141812A1 (en) * | 2019-05-24 | 2020-12-03 | City Of Hope | Ccr4 targeted chimeric antigen receptor modified t cells for treatment of ccr4 positive malignancies |
CA3143108A1 (en) * | 2019-06-19 | 2020-12-24 | Julius-Maximilians-Universitat Wurzburg | Ultramodular igg3-based spacer domain and multi-function site for implementation in chimeric antigen receptor design |
CN112442126B (zh) * | 2019-09-04 | 2022-07-29 | 杭州济元基因科技有限公司 | 一种抗人cs1抗原的单克隆抗体及其car-t细胞 |
JP2023525049A (ja) * | 2020-05-08 | 2023-06-14 | シアトル・チルドレンズ・ホスピタル・ドゥーイング/ビジネス/アズ・シアトル・チルドレンズ・リサーチ・インスティテュート | ナチュラルキラー細胞を標的とするキメラ抗原受容体(car) |
CN114286691A (zh) * | 2020-08-06 | 2022-04-05 | 武汉思安医疗技术有限公司 | Cs1-抗体和抗cs1-car-t细胞 |
JP2024500847A (ja) | 2020-12-18 | 2024-01-10 | センチュリー セラピューティクス,インコーポレイテッド | 適合可能な受容体特異性を有するキメラ抗原受容体システム |
CN117083292A (zh) * | 2021-04-02 | 2023-11-17 | 科济生物医药(上海)有限公司 | Cs1工程化细胞及其组合物 |
WO2023107898A1 (en) * | 2021-12-06 | 2023-06-15 | The Children's Hospital Of Philadelphia | Dual targeting of pediatric malignancies through car t-cells secreting bispecific innate immune cell engagers (bices) |
CN116410331B (zh) * | 2021-12-31 | 2024-01-30 | 合源生物科技(天津)有限公司 | 靶向cs1的嵌合抗原受体、靶向bcma/cs1的双特异性嵌合抗原受体及其应用 |
WO2023180511A1 (en) * | 2022-03-25 | 2023-09-28 | F. Hoffmann-La Roche Ag | Improved chimeric receptors |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101313069A (zh) * | 2005-11-24 | 2008-11-26 | Ucb医药有限公司 | 生物测定法 |
WO2013123061A1 (en) * | 2012-02-13 | 2013-08-22 | Seattle Children's Hospital D/B/A Seattle Children's Research Institute | Bispecific chimeric antigen receptors and therapeutic uses thereof |
WO2014179759A1 (en) * | 2013-05-03 | 2014-11-06 | Ohio State Innovation Foundation | Cs1-specific chimeric antigen receptor engineered immune effector cells |
Family Cites Families (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SG59954A1 (en) | 1989-08-11 | 1999-02-22 | Amrad Corp Ltd | Improvements in granulocyte-macrophage colony- stimulating factor receptor and derivatives thereof |
WO2006074399A2 (en) * | 2005-01-05 | 2006-07-13 | Biogen Idec Ma Inc. | Multispecific binding molecules comprising connecting peptides |
MY169746A (en) | 2005-08-19 | 2019-05-14 | Abbvie Inc | Dual variable domain immunoglobulin and uses thereof |
WO2007047829A2 (en) | 2005-10-19 | 2007-04-26 | Laboratoires Serono S.A. | Novel heterodimeric proteins and uses thereof |
MX2008015524A (es) * | 2006-06-12 | 2009-01-13 | Trubion Pharmaceuticals Inc | Proteinas de union multivalentes monocatenarias con funcion efectora. |
CN102844442B (zh) | 2010-02-12 | 2016-09-07 | 昂考梅德药品有限公司 | 用于鉴定和分离表达多肽的细胞的方法 |
JP5947311B2 (ja) | 2010-12-09 | 2016-07-06 | ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア | 癌を治療するためのキメラ抗原受容体改変t細胞の使用 |
EP3421489B1 (en) | 2012-03-23 | 2021-05-05 | The United States of America, as represented by The Secretary, Department of Health and Human Services | Anti-mesothelin chimeric antigen receptors |
KR102132041B1 (ko) | 2012-04-05 | 2020-07-09 | 에이씨 이뮨 에스.에이. | 인간화된 타우 항체 |
CA2936501A1 (en) * | 2014-01-13 | 2015-07-16 | Stephen J. Forman | Chimeric antigen receptors (cars) having mutations in the fc spacer region and methods for their use |
PT3105317T (pt) * | 2014-02-14 | 2019-02-27 | Cellectis | Células para imunoterapia manipuladas para atuar sobre antigénios presentes tanto em células imunitárias como em células patológicas |
AU2015254595B2 (en) * | 2014-05-02 | 2019-06-27 | Cellectis | CS1 specific multi-chain chimeric antigen receptor |
CN107429253B (zh) | 2014-12-05 | 2021-11-05 | 希望之城公司 | Cs1靶向性嵌合抗原受体修饰的t细胞 |
BR112019019606A2 (pt) * | 2017-03-20 | 2020-04-14 | Hope City | células t modificadas pelo receptor de antígeno quimérico direcionado a cs1 |
-
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101313069A (zh) * | 2005-11-24 | 2008-11-26 | Ucb医药有限公司 | 生物测定法 |
WO2013123061A1 (en) * | 2012-02-13 | 2013-08-22 | Seattle Children's Hospital D/B/A Seattle Children's Research Institute | Bispecific chimeric antigen receptors and therapeutic uses thereof |
WO2014179759A1 (en) * | 2013-05-03 | 2014-11-06 | Ohio State Innovation Foundation | Cs1-specific chimeric antigen receptor engineered immune effector cells |
Non-Patent Citations (2)
Title |
---|
Genetic Modification of T Cells Redirected toward CS1 Enhances Eradication of Myeloma Cells;Jianhong Chu等;《Clin Cancer Res》;20140327;第20卷(第15期);"Materials and Methods"部分 * |
Jianhong Chu等.Genetic Modification of T Cells Redirected toward CS1 Enhances Eradication of Myeloma Cells.《Clin Cancer Res》.2014,第20卷(第15期), * |
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