{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T09:06:13Z","timestamp":1788339973050,"version":"build-2803163510"},"reference-count":46,"publisher":"BMJ","issue":"3","license":[{"start":{"date-parts":[[2021,3,19]],"date-time":"2021-03-19T00:00:00Z","timestamp":1616112000000},"content-version":"unspecified","delay-in-days":18,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["J Immunother Cancer"],"accepted":{"date-parts":[[2021,1,28]]},"published-print":{"date-parts":[[2021,3]]},"abstract":"<jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Tumors often develop resistance to surveillance by endogenous immune cells, which include natural killer (NK) cells. Ex vivo activated and\/or expanded NK cells demonstrate cytotoxicity against various tumor cells and are promising therapeutics for adoptive cancer immunotherapy. Genetic modification can further enhance NK effector cell activity or activation sensitization. Here, we evaluated the effect of the genetic deletion of ubiquitin ligase Casitas B-lineage lymphoma pro-oncogene-b (<jats:italic>CBLB<\/jats:italic>), a negative regulator of lymphocyte activity, on placental CD34<jats:sup>+<\/jats:sup> cell-derived NK (PNK) cell cytotoxicity against tumor cells.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>Using CRISPR\/Cas9 technology, <jats:italic>CBLB<\/jats:italic> was knocked out in placenta-derived CD34<jats:sup>+<\/jats:sup> hematopoietic stem cells, followed by differentiation into PNK cells. Cell expansion, phenotype and cytotoxicity against tumor cells were characterized in vitro. The antitumor efficacy of <jats:italic>CBLB<\/jats:italic> knockout (KO) PNK cells was tested in an acute myeloid leukemia (HL-60) tumor model in NOD-<jats:italic>scid<\/jats:italic> IL2R gamma<jats:sup>null<\/jats:sup> (NSG) mice. PNK cell persistence, biodistribution, proliferation, phenotype and antitumor activity were evaluated.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>94% of <jats:italic>CBLB<\/jats:italic> KO efficacy was achieved using CRISPR\/Cas9 gene editing technology. <jats:italic>CBLB<\/jats:italic> KO placental CD34<jats:sup>+<\/jats:sup> cells differentiated into PNK cells with high cell yield and &gt;90% purity determined by CD56<jats:sup>+<\/jats:sup> CD3<jats:sup>\u2212<\/jats:sup> cell identity. Ablation of <jats:italic>CBLB<\/jats:italic> did not impact cell proliferation, NK cell differentiation or phenotypical characteristics of PNK cells. When compared with the unmodified PNK control, <jats:italic>CBLB<\/jats:italic> KO PNK cells exhibited higher cytotoxicity against a range of liquid and solid tumor cell lines in vitro. On infusion into busulfan-conditioned NSG mice, <jats:italic>CBLB<\/jats:italic> KO PNK cells showed in vivo proliferation and maturation as evidenced by increased expression of CD16, killer Ig-like receptors and NKG2A over 3 weeks. Additionally, <jats:italic>CBLB<\/jats:italic> KO PNK cells showed greater antitumor activity in a disseminated HL60-luciferase mouse model compared with unmodified PNK cells.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>Conclusion<\/jats:title>\n                  <jats:p>\n                     <jats:italic>CBLB<\/jats:italic> ablation increased PNK cell effector function and proliferative capacity compared with non-modified PNK cells. These data suggest that targeting <jats:italic>CBLB<\/jats:italic> may offer therapeutic advantages via enhancing antitumor activities of NK cell therapies.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1136\/jitc-2020-001975","type":"journal-article","created":{"date-parts":[[2021,3,19]],"date-time":"2021-03-19T12:17:40Z","timestamp":1616156260000},"page":"e001975","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":36,"title":["<i>CBLB<\/i> ablation with CRISPR\/Cas9 enhances cytotoxicity of human placental stem cell-derived NK cells for cancer immunotherapy"],"prefix":"10.1136","volume":"9","author":[{"given":"Xuan","family":"Guo","sequence":"first","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9059-0486","authenticated-orcid":false,"given":"Tanel","family":"Mahlak\u00f5iv","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qian","family":"Ye","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Srinivas","family":"Somanchi","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuyang","family":"He","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hemlata","family":"Rana","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"DiFiglia","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph","family":"Gleason","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William","family":"van der Touw","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Hariri","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaokui","family":"Zhang","sequence":"additional","affiliation":[{"name":"Celularity Inc, Florham Park, New Jersey, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"239","published-online":{"date-parts":[[2021,3,19]]},"reference":[{"key":"2025100308505391000_9.3.e001975.1","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1038\/s41573-019-0052-1","article-title":"Nk cells for cancer immunotherapy","volume":"19","author":"Shimasaki","year":"2020","journal-title":"Nat Rev Drug Discov"},{"key":"2025100308505391000_9.3.e001975.2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2015.00266"},{"key":"2025100308505391000_9.3.e001975.3","doi-asserted-by":"crossref","unstructured":"Vivier E , Tomasello E , Baratin M . Functions of natural killer cells. 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Cbl-B silenced human NK cells respond stronger to cytokine stimulation. J Immunother Cancer 2015;3.doi:10.1186\/2051-1426-3-S2-P230","DOI":"10.1186\/2051-1426-3-S2-P230"},{"key":"2025100308505391000_9.3.e001975.33","doi-asserted-by":"crossref","DOI":"10.3389\/fimmu.2013.00101","article-title":"Characterization and ex vivo expansion of human placenta-derived natural killer cells for cancer immunotherapy","volume":"4","author":"Kang","year":"2013","journal-title":"Front Immunol"},{"key":"2025100308505391000_9.3.e001975.34","doi-asserted-by":"crossref","unstructured":"Cooley S , Hari P , McCloskey J . Abstract CT079: a phase I study of PNK-007, allogeneic, off the shelf NK cell in relapsed\/refractory AML (NCT02781467) 2019.doi:10.1158\/1538-7445.AM2019-CT079","DOI":"10.1158\/1538-7445.AM2019-CT079"},{"key":"2025100308505391000_9.3.e001975.35","doi-asserted-by":"crossref","unstructured":"Holstein SA , Cooley SA , Hari P . 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Nk cell-based immunotherapies in cancer. 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