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HuM195 and its single-chain variable fragment increase Aβ phagocytosis in microglia via elimination of CD33 inhibitory signaling

Molecular Psychiatry (2024)Cite this article

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Abstract

CD33 is a transmembrane receptor expressed on cells of myeloid lineage and regulates innate immunity. CD33 is a risk factor for Alzheimer’s disease (AD) and targeting CD33 has been a promising strategy drug development. However, the mechanism of CD33’s action is poorly understood. Here we investigate the mechanism of anti-CD33 antibody HuM195 (Lintuzumab) and its single-chain variable fragment (scFv) and examine their therapeutic potential. Treatment with HuM195 full-length antibody or its scFv increased phagocytosis of β-amyloid 42 (Aβ42) in human microglia and monocytes. This activation of phagocytosis was driven by internalization and degradation of CD33, thereby downregulating its inhibitory signal. HumM195 transiently induced CD33 phosphorylation and its signaling via receptor dimerization. However, this signaling decayed with degradation of CD33. scFv binding to CD33 leads to a degradation of CD33 without detection of the CD33 dimerization and signaling. Moreover, we found that treatments with either HuM195 or scFv promotes the secretion of IL33, a cytokine implicated in microglia reprogramming. Importantly, recombinant IL33 potentiates the uptake of Aβ42 in monocytes. Collectively, our findings provide unanticipated mechanistic insight into the role of CD33 signaling in both monocytes and microglia and define a molecular basis for the development of CD33-based therapy of AD.

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Fig. 1: Endotoxin free purified Aβ peptides does not stimulate immune reaction and have homogeneous characteristics of Aβ oligomers.
Fig. 2: HuM195 and its single-chain variable fragment (scFv) treatment increase Aβ42 uptake by phagocytic cells through CD33 degradation.
Fig. 3: HuM195 IgG but not HuM195 scFv incubation induces CD33 dimerization and phosphorylation, relaying transient downstream signaling.
Fig. 4: HuM195 treatment induces expression of immune-related genes including IL33 which enhance phagocytic ability.
Fig. 5: The mechanism of HuM195 treatment in AD: induces microglia activation and modifies inflammatory response.

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Data availability

Source data for Figs. 1–5 and Extended Figs. 1–5 and Tables S1 and S2, as well as all other data, are available from the corresponding authors upon reasonable request.

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Funding

This work is supported by NIH grant RF1AG057593 (YML), R01AG061350 (YML), R01AG080684 (YML). the JPB Foundation (YML), the MetLife Foundation (YML), Cure Alzheimer’s Fund (YML), The Edward and Della L. Thome Memorial Foundation (YML), Coins for the Alzheimer’s Research Trust (YML) and BrightFocus Foundation (EW), R35 CA241894 (DAS), R01 CA055349 (DAS), P01 CA023766 (DAS), Tudor Funds (DAS). Authors also acknowledge the MSK Cancer Center Support Grant/Core Grant (Grant P30 CA008748), Mr. William H. Goodwin and Mrs. Alice Goodwin and the Commonwealth Foundation for Cancer Research, the Experimental Therapeutics Center of MSKCC, and the William Randolph Hearst Fund in Experimental Therapeutics.

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  1. These authors contributed equally: Eitan Wong, Manish Malviya.

Authors and Affiliations

  1. Chemical Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, 10065, USA

    Eitan Wong, Tanya Jain, George P. Liao, Zoe Kehs & Yue-Ming Li

  2. Molecular Pharmacology Program, Memorial Sloan-Kettering Cancer Center, New York, NY, 10065, USA

    Manish Malviya & David A. Scheinberg

  3. Program of Neurosciences, Weill Graduate School of Medical Sciences of Cornell University, New York, NY, 10021, USA

    Tanya Jain & Yue-Ming Li

  4. Program of Pharmacology Weill Graduate School of Medical Sciences of Cornell University, New York, NY, 10021, USA

    George P. Liao, Zoe Kehs, David A. Scheinberg & Yue-Ming Li

  5. Laboratory of Molecular and Cellular Neuroscience, The Rockefeller University, New York, NY, 10021, USA

    Jerry C. Chang

  6. Developmental biology program, Memorial Sloan-Kettering Cancer Center, New York, NY, 10021, USA

    Lorenz Studer

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Contributions

EW, DAS and YML conceived the study. EW, MM, DAS and YML planned and EW, MM performed most of the experiments and analyzed the data. TJ prepared ES microglia, GPL and ZK expressed and purified Huam195 scFv. JCC performed AFM studies. LS supervised the production of ES microglia and analyzed data. EW wrote the initial draft of the manuscript and EW, MM, DAS and YML revised the manuscript. All authors discussed and commented on the manuscript.

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Correspondence to David A. Scheinberg or Yue-Ming Li.

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Wong, E., Malviya, M., Jain, T. et al. HuM195 and its single-chain variable fragment increase Aβ phagocytosis in microglia via elimination of CD33 inhibitory signaling. Mol Psychiatry (2024). https://doi.org/10.1038/s41380-024-02474-z

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