An arrayed CRISPR screen of primary B cells reveals the essential elements of the antibody secretion pathway

Stephanie Trezise; Isabella Y Kong; Edwin D Hawkins; Marco J Herold; Simon N Willis; Stephen L Nutt

doi:10.3389/fimmu.2023.1089243

An arrayed CRISPR screen of primary B cells reveals the essential elements of the antibody secretion pathway

Front Immunol. 2023 Feb 13:14:1089243. doi: 10.3389/fimmu.2023.1089243. eCollection 2023.

Authors

Stephanie Trezise^{1

2

3}, Isabella Y Kong^{1

2

4}, Edwin D Hawkins^{1

2}, Marco J Herold^{1

2}, Simon N Willis^{1

2}, Stephen L Nutt^{1

2}

Affiliations

¹ Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia.
² Department of Medical Biology, The University of Melbourne, Parkville, VIC, Australia.
³ Center for Immunology and Inflammatory Diseases, Massachusetts General Hospital, Harvard Medical School, Harvard University, Boston, MA, United States.
⁴ Department of Pediatrics, Division of Pediatric Hematology/Oncology, Weill Cornell Medicine, New York, NY, United States.

Abstract

Background: Humoral immunity depends on the differentiation of B cells into antibody secreting cells (ASCs). Excess or inappropriate ASC differentiation can lead to antibody-mediated autoimmune diseases, while impaired differentiation results in immunodeficiency.

Methods: We have used CRISPR/Cas9 technology in primary B cells to screen for regulators of terminal differentiation and antibody production.

Results: We identified several new positive (Sec61a1, Hspa5) and negative (Arhgef18, Pold1, Pax5, Ets1) regulators that impacted on the differentiation process. Other genes limited the proliferative capacity of activated B cells (Sumo2, Vcp, Selk). The largest number of genes identified in this screen (35) were required for antibody secretion. These included genes involved in endoplasmic reticulum-associated degradation and the unfolded protein response, as well as post-translational protein modifications.

Discussion: The genes identified in this study represent weak links in the antibody-secretion pathway that are potential drug targets for antibody-mediated diseases, as well as candidates for genes whose mutation results in primary immune deficiency.

Keywords: ER associated degradation (ERAD); endoplasmic reticulum; humoral immunity; immunodeficiency; in vitro differentiation; plasma cell; unfolded protein response.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies
B-Lymphocytes
Endoplasmic Reticulum-Associated Degradation*
Immunity, Humoral
Secretory Pathway*

Substances

Antibodies

Grants and funding

This work was supported by the National Health and Medical Research Council (NHMRC) of Australia (1144905 and 11155342 to SN, 1159658,1186575, 1145728 and 1156095 to MH), the Walter and Eliza Hall Trust (to SW), Cancer Council Victoria (1147328 and 2021 Grant in Aid to MH). This work was made possible through Victorian State Government Operational Infrastructure Support and NHMRC Independent Research Institutes Infrastructure Support.