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Here, cell surface proteomics revealed CD72 as an optimal target for poor-prognosis KMT2A\/MLL1-rearranged (MLLr) B-cell acute lymphoblastic leukemia (B-ALL), which we further found to be expressed in other B-cell malignancies. Using a recently described, fully in vitro system, we selected synthetic CD72-specific nanobodies, incorporated them into chimeric antigen receptors (CAR), and demonstrated robust activity against B-cell malignancy models, including CD19 loss. Taking advantage of the role of CD72 in inhibiting B-cell receptor signaling, we found that SHIP1 inhibition increased CD72 surface density. We establish that CD72-nanobody CAR-T cells are a promising therapy for MLLr B-ALL.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Significance:<\/jats:title>\n                    <jats:p>Patients with MLLr B-ALL have poor prognoses despite recent immunotherapy advances. Here, surface proteomics identifies CD72 as being enriched on MLLr B-ALL but also widely expressed across B-cell cancers. We show that a recently described, fully in vitro nanobody platform generates binders highly active in CAR-T cells and demonstrate its broad applicability for immunotherapy development.<\/jats:p>\n                    <jats:p>This article is highlighted in the In This Issue feature, p. 1861<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1158\/2159-8290.cd-20-0242","type":"journal-article","created":{"date-parts":[[2021,3,16]],"date-time":"2021-03-16T15:50:33Z","timestamp":1615909833000},"page":"2032-2049","update-policy":"https:\/\/doi.org\/10.1158\/crossmark_policy","source":"Crossref","is-referenced-by-count":73,"title":["Surface Proteomics Reveals CD72 as a Target for\n                    <i>In Vitro<\/i>\n                    \u2013Evolved Nanobody-Based CAR-T Cells in\n                    <i>KMT2A\/MLL1<\/i>\n                    -Rearranged B-ALL"],"prefix":"10.1158","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7741-5368","authenticated-orcid":false,"given":"Matthew A.","family":"Nix","sequence":"first","affiliation":[{"name":"1Department of Laboratory Medicine, University of California, San Francisco, San Francisco, California."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kamal","family":"Mandal","sequence":"additional","affiliation":[{"name":"1Department of Laboratory Medicine, University of California, San Francisco, San Francisco, California."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huimin","family":"Geng","sequence":"additional","affiliation":[{"name":"1Department of Laboratory Medicine, University of California, San Francisco, San Francisco, California."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neha","family":"Paranjape","sequence":"additional","affiliation":[{"name":"1Department of Laboratory Medicine, University of California, 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California."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan","family":"Ramirez","sequence":"additional","affiliation":[{"name":"4Cardiovascular Research Institute, University of California, San Francisco, San Francisco, California."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"Deucher","sequence":"additional","affiliation":[{"name":"1Department of Laboratory Medicine, University of California, San Francisco, San Francisco, California."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Phojanokong","sequence":"additional","affiliation":[{"name":"5Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2499-6618","authenticated-orcid":false,"given":"Veronica","family":"Steri","sequence":"additional","affiliation":[{"name":"5Helen Diller Family 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