Department of Pathology, Amsterdam UMC location University of Amsterdam, Meibergdreef 9, Amsterdam, The Netherlands.
Lymphoma and Myeloma Center Amsterdam (LYMMCARE), Amsterdam, The Netherlands.
Nat Commun. 2022 Apr 19;13(1):2136. doi: 10.1038/s41467-022-29835-y.
The clinical introduction of the Bruton's tyrosine kinase (BTK) inhibitor ibrutinib, which targets B-cell antigen-receptor (BCR)-controlled integrin-mediated retention of malignant B cells in their growth-supportive lymphoid organ microenvironment, provided a major breakthrough in lymphoma and leukemia treatment. Unfortunately, a significant subset of patients is intrinsically resistant or acquires resistance against ibrutinib. Here, to discover novel therapeutic targets, we present an unbiased loss-of-adhesion CRISPR-Cas9 knockout screening method to identify proteins involved in BCR-controlled integrin-mediated adhesion. Illustrating the validity of our approach, several kinases with an established role in BCR-controlled adhesion, including BTK and PI3K, both targets for clinically applied inhibitors, are among the top hits of our screen. We anticipate that pharmacological inhibitors of the identified targets, e.g. PAK2 and PTK2B/PYK2, may have great clinical potential as therapy for lymphoma and leukemia patients. Furthermore, this screening platform is highly flexible and can be easily adapted to identify cell adhesion-regulatory proteins and signaling pathways for other stimuli, adhesion molecules, and cell types.
布鲁顿酪氨酸激酶（BTK）抑制剂伊布替尼的临床应用为淋巴瘤和白血病的治疗带来了重大突破，该抑制剂靶向 B 细胞抗原受体（BCR）控制的整合素介导的恶性 B 细胞在其生长支持性淋巴器官微环境中的保留。不幸的是，相当一部分患者对伊布替尼具有内在耐药性或获得了耐药性。在这里，为了发现新的治疗靶点，我们采用了一种无偏倚的去黏附 CRISPR-Cas9 敲除筛选方法，以鉴定参与 BCR 控制的整合素介导的黏附的蛋白质。该方法通过鉴定几种在 BCR 控制的黏附中具有既定作用的激酶（包括 BTK 和 PI3K，它们都是临床应用抑制剂的靶点）作为我们筛选的重要靶点，证明了其有效性。我们预计，所鉴定的靶标的药理学抑制剂，例如 PAK2 和 PTK2B/PYK2，可能作为淋巴瘤和白血病患者的治疗方法具有巨大的临床潜力。此外，该筛选平台具有高度的灵活性，可以轻松适应其他刺激、黏附分子和细胞类型，用于鉴定细胞黏附调节蛋白和信号通路。
