Dissecting phenotypic transitions in metastatic disease via photoconversion-based isolation.
Dissecting phenotypic transitions in metastatic disease via photoconversion-based isolation.
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DOI：
10.7554/elife.63270
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发表时间：
2021-02-23
期刊：
影响因子：
7.7
通讯作者：
Stanger BZ
中科院分区：
文献类型：
--
作者：
Sela Y;Li J;Kuri P;Merrell AJ;Li N;Lengner C;Rompolas P;Stanger BZ
癌症患者经常有隐匿性转移，这是复发的潜在来源，只能通过全身治疗。由于缺乏在空间上分离离散细胞的工具，对这一神秘部分的研究受到了限制。我们开发了PIC-IT，这是一种基于光转换的分离技术，可以有效地恢复任何大小的细胞团-包括单转移细胞-这些细胞团在很大程度上是无法获得的。在小鼠胰腺癌模型中，自发产生的微菌落的转录谱显示表型异质性，功能上的增殖倾向降低和与NF-κB/AP-1信号相关的炎症反应表型的富集。NF-κB的药理抑制减少了微菌落，但对大转移没有影响，这表明微菌落特别依赖于这一途径。PIC-IT因此能够系统地研究转移异质性。此外，该技术可以应用于其他生物系统，其中分离和表征空间上不同的细胞群目前是不可行的。
Cancer patients often harbor occult metastases, a potential source of relapse that is targetable only through systemic therapy. Studies of this occult fraction have been limited by a lack of tools with which to isolate discrete cells on spatial grounds. We developed PIC-IT, a photoconversion-based isolation technique allowing efficient recovery of cell clusters of any size – including single-metastatic cells – which are largely inaccessible otherwise. In a murine pancreatic cancer model, transcriptional profiling of spontaneously arising microcolonies revealed phenotypic heterogeneity, functionally reduced propensity to proliferate and enrichment for an inflammatory-response phenotype associated with NF-κB/AP-1 signaling. Pharmacological inhibition of NF-κB depleted microcolonies but had no effect on macrometastases, suggesting microcolonies are particularly dependent on this pathway. PIC-IT thus enables systematic investigation of metastatic heterogeneity. Moreover, the technique can be applied to other biological systems in which isolation and characterization of spatially distinct cell populations is not currently feasible.