Kinobead Profiling Reveals Reprogramming of BCR Signaling in Response to Therapy within Primary CLL Cells

Linley, Adam; Karydis, Laura; Mondru, Anil Kumar; D’Avola, Annalisa; Shmrany, Humood Al; Cicconi, Silvia; Griffin, Rebecca; Forconi, Francesco; Pettitt, Andrew R.; Kalakonda, Nagesh; Rawstron, Andrew; Hillmen, Peter; Steele, Andrew J.; MacEwan, David J.; Packham, Graham; Prior, Ian A.; Slupsky, Joseph R.

doi:10.1158/1078-0432.ccr-21-0161

Cited by 5 publications

(3 citation statements)

References 51 publications

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“…To understand how BCR signaling is affected by mutations such as BTK L528W that impair kinase function, we next used a kinobead/mass spectrometry–based protocol to systematically evaluate the abundance of kinases and their active site availability in BTK WT and mutant TMD8 cells. Recently, this approach was successfully used to probe BCR signaling in CLL cells ( 32 ) and was performed here with and without IgM stimulation and in the presence or absence of pirtobrutinib treatment (fig. S1G).…”

Section: Drug Resistance Mutations In Btk Have Distinct Enzymatic Act...mentioning

confidence: 99%

Kinase-impaired BTK mutations are susceptible to clinical-stage BTK and IKZF1/3 degrader NX-2127

Montoya,

Bourcier,

Noviski

et al. 2024

Science

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Increasing use of covalent and noncovalent inhibitors of Bruton’s tyrosine kinase (BTK) has elucidated a series of acquired drug-resistant BTK mutations in patients with B cell malignancies. Here we identify inhibitor resistance mutations in BTK with distinct enzymatic activities, including some that impair BTK enzymatic activity while imparting novel protein-protein interactions that sustain B cell receptor (BCR) signaling. Furthermore, we describe a clinical-stage BTK and IKZF1/3 degrader, NX-2127, that can bind and proteasomally degrade each mutant BTK proteoform, resulting in potent blockade of BCR signaling. Treatment of chronic lymphocytic leukemia with NX-2127 achieves >80% degradation of BTK in patients and demonstrates proof-of-concept therapeutic benefit. These data reveal an oncogenic scaffold function of mutant BTK that confers resistance across clinically approved BTK inhibitors but is overcome by BTK degradation in patients.

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Section: Drug Resistance Mutations In Btk Have Distinct Enzymatic Act...mentioning

confidence: 99%

Kinase-impaired BTK mutations are susceptible to clinical-stage BTK and IKZF1/3 degrader NX-2127

Montoya,

Bourcier,

Noviski

et al. 2024

Science

View full text Add to dashboard Cite

show abstract

“…This study revealed that the signature of isolated kinases in CLL cells from pre‐treatment patients is significantly different from patients receiving chemoimmunotherapy and ibrutinib treatment and that individual patients also showed signalling heterogeneity in response to the therapy [85]. It was also found that the CLL cells from ibrutinib‐treated patients undergo active rewiring of BCR signalling with significant increase activation of SYK, FAK2, GAK and RSK2 [85]. This study hence demonstrates that the kinobeads approach could provide insights into the potential drivers of signal pathway reprogramming in response to the therapy that may inform further treatment options.…”

Section: Applications Of Mass Spectrometry‐based Proteomics To Study ...mentioning

confidence: 99%

“…Recently, Linley et al [85] applied a kinobeads‐based approach to study the adaptive response of the B‐cell receptor (BCR) signalling in primary CLL cells following exposure to ibrutinib. This study revealed that the signature of isolated kinases in CLL cells from pre‐treatment patients is significantly different from patients receiving chemoimmunotherapy and ibrutinib treatment and that individual patients also showed signalling heterogeneity in response to the therapy [85]. It was also found that the CLL cells from ibrutinib‐treated patients undergo active rewiring of BCR signalling with significant increase activation of SYK, FAK2, GAK and RSK2 [85].…”

Section: Applications Of Mass Spectrometry‐based Proteomics To Study ...mentioning

confidence: 99%

A recent update on small‐molecule kinase inhibitors for targeted cancer therapy and their therapeutic insights from mass spectrometry‐based proteomic analysis

2022

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Kinases are key regulatory signalling proteins governing numerous essential biological processes and cellular functions. Dysregulation of many protein kinases is associated with cancer initiation and progression. Given their crucial roles, there has been increasing interest in harnessing kinases as prospective drug targets for cancer. In recent decades, numerous small‐molecule kinase inhibitors have been developed and revolutionized the cancer treatment landscape. Despite their great potential, challenges remain in developing highly selective and effective kinase inhibitors, with toxicity and resistance issues frequently arising. In this review, we first provide an overview of the role of kinases in carcinogenesis and describe the current progress with small‐molecule kinase inhibitors that have been approved for clinical use. We then discuss the application of mass spectrometry (MS)‐based proteomics strategies to help in the design of kinase inhibitors. Finally, we discuss the challenges and outlook concerning MS‐based proteomics techniques for kinase drug research.

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Bruton's tyrosine kinase inhibition induces rewiring of proximal and distal B‐cell receptor signaling in mice

et al. 2021

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Bruton s tyrosine kinase (Btk) is a crucial signaling molecule in BCR signaling and a key regulator of B-cell differentiation and function. Btk inhibition has shown impressive clinical efficacy in various B-cell malignancies. However, it remains unknown whether inhibition additionally induces changes in BCR signaling due to feedback mechanisms, a phenomenon referred to as BCR rewiring. In this report, we studied the impact of Btk activity on major components of the BCR signaling pathway in mice. As expected, NF-κB and Akt/S6 signaling was decreased in Btk-deficient B cells. Unexpectedly, phosphorylation of several proximal signaling molecules, including CD79a, Syk, and PI3K, as well as the key Btk-effector PLCγ2 and the more downstream kinase Erk, were significantly increased. This pattern of BCR rewiring was essentially opposite in B cells from transgenic mice overexpressing Btk. Importantly, prolonged Btk inhibitor treatment of WT mice or mice engrafted with leukemic B cells also resulted in increased phosho-CD79a and phospho-PLCγ2 in B cells. Our findings show that Btk enzymatic function determines phosphorylation of proximal and distal BCR signaling molecules in B cells. We conclude that Btk inhibitor treatment results in rewiring of BCR signaling, which may affect both malignant and healthy B cells.

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Kinobead Profiling Reveals Reprogramming of BCR Signaling in Response to Therapy within Primary CLL Cells

Abstract: Running title: Therapy induces adaptive signaling changes in primary CLL.

Cited by 5 publications

References 51 publications

Kinase-impaired BTK mutations are susceptible to clinical-stage BTK and IKZF1/3 degrader NX-2127

Kinase-impaired BTK mutations are susceptible to clinical-stage BTK and IKZF1/3 degrader NX-2127

A recent update on small‐molecule kinase inhibitors for targeted cancer therapy and their therapeutic insights from mass spectrometry‐based proteomic analysis

Bruton's tyrosine kinase inhibition induces rewiring of proximal and distal B‐cell receptor signaling in mice

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