Glycoengineering of NK cells with glycan ligands of CD22 and selectins for B-cell lymphoma therapy

Abstract

CD22, a member of Siglec family of sialic acid binding proteins, has restricted expression on B cells. Antibody-based agents targeting CD22 or CD20 (Rituxan TM ) on B lymphoma and leukemia cells exhibit clinical efficacy for treating these malignancies, but also attack normal B cells leading to immune deficiency. Here, we report a chemoenzymatic glycocalyx editing strategy to introduce high-affinity and specific CD22 ligands onto NK-92MI and cytokine-induced killer (CIK) cells to achieve tumor-specific CD22 targeting. These CD22-ligand modified cells exhibited significantly enhanced tumor cell binding and killing in vitro without harming healthy B cells. For effective lymphoma cell killing in vivo we further functionalized CD22 ligand-modified NK-92MI cells with the E-selectin ligand sialyl Lewis X to promote trafficking to bone marrow. The cells containing the ligands of both CD22 and selectins resulted in the efficient suppression of B lymphoma in a xenograft model.Our results suggest that NK cells modified with glycan ligands to CD22 and selectins promote both targeted killing of B lymphoma cells and improved trafficking to sites where the cancer cells reside, respectively.

Cell‐specific targeting of extracellular vesicles through engineering the glycocalyx

“…Indeed, rational glycoengineering has shown promises to enhance the therapeutic efficacy of adoptive cell therapies. For instance, natural killer cells displaying the glycan ligand of CD22 had higher affinity and activity against leukemic B cells than non‐targeted cells (Hong et al., 2020). In terms of glycoengineering of EVs, two earlier studies trimmed surface glycans using enzymatic digestion and found significant changes in cellular uptake and biodistribution profiles, highlighting the functional relevance of surface glycans (Royo et al., 2019; Williams et al., 2019).…”

Modulation of Siglec-7 Signaling Via In Situ-Created High-Affinity cis-Ligands

“…If so, this strategy may serve as a general approach to downregulate Siglecs’ inhibitory functions. In this endeavor, we have already demonstrated that high-affinity and specific ligands for other Siglecs, such as Siglec-2 (CD22) 53 and Siglec-9, 9 , 41 , 54 can be introduced onto live cells by employing similar strategies ( Supporting Information Figures S26 and S28 ). A transplantation of expanded allogeneic NK cells has emerged as a promising strategy for cancer treatment; 55 − 57 we envisage that, by modifying these NK cells with the high-affinity ligands prior to adoptive transfer, the constitutively expressed Siglec-7 could be released to enhance the NK effector functions for a better antitumor immunity.…”

Genetic glycoengineering in mammalian cells

“…The unique functions of several other GALNT isoenzymes were also explored in KO studies (153)(154)(155)(156)(157)(158)(159). Moreover, glycoengineering is being used with approaches for metabolic labeling and tagging of glycosylation (22,23,(160)(161)(162), including the azido sugars for click chemistry introduced by the Bertozzi group (163)(164)(165). Glycoengineered cells allow for screening and validation of selective binding and labeling probes in live cells as exemplified by the bump-and-hole strategy employed by Schumann and colleagues (166), in which modified UDP-GalNAc donor substrates (bumped) and GALNTs engineered to selectively accommodate these by an enlarged active site (hole) are used to detect isoform-specific functions in transfected cells (167)(168)(169).…”