huHSC-NCG小鼠是將人造血干細(xì)胞 (CD34+HSC）移植到輻照清髓的重度免疫缺陷小鼠NCG體內(nèi)，并分化產(chǎn)生各類造血或者免疫細(xì)胞，如T細(xì)胞、B細(xì)胞以及NK細(xì)胞等，從而獲得免疫系統(tǒng)人源化的模型以NCG小鼠為受體鼠，構(gòu)建huHSC-NCG人源化小鼠。

應(yīng)用領(lǐng)域

1. 多種腫瘤免疫治療藥物的評(píng)價(jià)

2. 免疫相關(guān)的藥物評(píng)價(jià)

模型數(shù)據(jù)

圖1. NCG小鼠清髓效果檢測(cè)

由于輻照可以清除小鼠骨髓中的造血干細(xì)胞，從而提高人源HSC移植后的重建水平，因此選用輻照處理后的NCG小鼠移植人源HSC。利用流式細(xì)胞技術(shù)檢測(cè)輻照清髓效果。其中mCD45為小鼠白細(xì)胞標(biāo)記物，mCD117為造血干細(xì)胞標(biāo)記物。上圖顯示輻照后mCD45+mCD117+細(xì)胞群體比例為1.11%,顯著低于未輻照組（13.5%）。表明150 cGys劑量輻照6-7周NCG小鼠可達(dá)到理想的清髓效果。

圖2. huHSC-NCG小鼠的生存曲線及體重變化情況

huHSC-NCG小鼠存活時(shí)間可超過(guò)37周，且生存率與NCG野生型小鼠無(wú)顯著差異。huHSC-NCG小鼠體重隨時(shí)間變化呈增長(zhǎng)趨勢(shì)，前期與NCG野生型小鼠體重?zé)o顯著差異，隨著時(shí)間延長(zhǎng)，huHSC-NCG小鼠體重下降，在實(shí)驗(yàn)終點(diǎn)，huHSC-NCG小鼠體重顯著低于NCG野生小鼠（***, P < 0.001）。數(shù)據(jù)以Mean ± SEM形式呈現(xiàn)。

圖3. huHSC-NCG重建效果鑒定

huHSC-NCG小鼠的免疫重建效率高，主要重建T細(xì)胞和大量未成熟的B細(xì)胞，以及少量NK細(xì)胞。

圖4. 基于huHSC-NCG小鼠模型的體內(nèi)藥效評(píng)價(jià)

基于huHSC-NCG小鼠體內(nèi)用藥試驗(yàn)。將對(duì)數(shù)生長(zhǎng)期人淋巴瘤細(xì)胞Raji細(xì)胞接種到huHSC-NCG小鼠皮下，待腫瘤生長(zhǎng)至平均體積約40-50 mm3時(shí)，根據(jù)小鼠腫瘤體積和體重，隨機(jī)分為Vehicle組，Tri-TE給藥組，Blincyto給藥組，并使用相應(yīng)的藥物進(jìn)行治療。結(jié)果顯示：Tri-TE組（TGI=62.04%）及Blincyto給藥組（TGI=51.14%）對(duì)huHSC-NCG的Raji細(xì)胞荷瘤鼠上腫瘤生長(zhǎng)有抑制作用。

NCG在CAR-T治療上的應(yīng)用

CAR-T（Chimeric Antigen Receptor T Cell）是腫瘤免疫治療的新方法。CAR-T細(xì)胞具有靶向識(shí)別腫瘤抗原的能力，其殺傷腫瘤細(xì)胞無(wú)需進(jìn)行抗原遞呈，與以往免疫療法相比，具有特異性高、攻擊持久等優(yōu)勢(shì)。在惡性腫瘤，特別是血液腫瘤治療中有很好的療效。集萃藥康具有豐富的CDX和自主知識(shí)產(chǎn)權(quán)的PDX庫(kù)，包括多種實(shí)體瘤和血液瘤，為CAR-T藥效評(píng)價(jià)提供了豐富的腫瘤模型資源。同時(shí)，集萃藥康開(kāi)發(fā)了外周血中CAR-T細(xì)胞計(jì)數(shù)方法和細(xì)胞因子檢測(cè)方法，滿足CAR-T實(shí)驗(yàn)檢測(cè)需求。

圖5.在NCG小鼠上接種Nalm6-Luciferase后，進(jìn)行CAR-T藥效評(píng)價(jià)

Nalm6-Luciferase細(xì)胞尾靜脈接種到NCG小鼠后，第7天根據(jù)腫瘤負(fù)荷進(jìn)行分組，分別給予Control T，CAR-T1及CAR-T2細(xì)胞治療，并于D12，D19，D26利用活體成像技術(shù)檢測(cè)腫瘤負(fù)荷。據(jù)體內(nèi)成像及統(tǒng)計(jì)結(jié)果顯示，與Control T相比，CAR-T1和CAR-T2細(xì)胞均能很好的抑制小鼠體內(nèi)腫瘤生長(zhǎng)，延長(zhǎng)小鼠生存期。

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