Potent Cytolytic Activity and Specific IL15 Delivery in a Second-Generation Trispecific Killer Engager

Felices, Martin; Lenvik, Todd; Kodal, Behiye; Hinderlie, Peter; Bendzick, Laura; Schirm, Dawn K.; Kaminski, Michael F.; McElmurry, Ron; Geller, Melissa A.; Eckfeldt, Craig E.; Vallera, Daniel A.; Miller, Jeffrey S.

doi:10.1158/2326-6066.cir-19-0837

Cited by 48 publications

(44 citation statements)

References 43 publications

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“…A recent upgrade to TriKEs is the replacement of the second tumor antigen-targeting scFv with IL15, as a way of increasing NK cell activation and expansion, simultaneously to promoting NK cell trafficking to tumors [150]. An anti-CD16 ×IL-15 × anti-CD33 TriKE is currently being tested in a phase I/II clinical trial (NCT03214666), and a second generation version has already been developed [151]. Other advancements include the development of: (i) a BIKE that additionally includes an anti-CD133 molecule to target cancer stem cells [152]; (ii) a TriKE that activates NK cells by binding NKp46 instead of presenting IL-15 [153]; and (iii) a nanoparticle-based TriKE that targets two NK cell activating receptors (CD16 and 4-1BB) and the tumor antigen EGFR, while simultaneously delivering the chemotherapeutic epirubicin [154].…”

Section: Antibody-based Constructsmentioning

confidence: 99%

Natural Killer Cells in Immunotherapy: Are We Nearly There?

Bachiller

Battram

Perez-Amill

et al. 2020

Cancers

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Natural killer (NK) cells are potent anti-tumor and anti-microbial cells of our innate immune system. They are equipped with a vast array of receptors that recognize tumor cells and other pathogens. The innate immune activity of NK cells develops faster than the adaptive one performed by T cells, and studies suggest an important immunoregulatory role for each population against the other. The association, observed in acute myeloid leukemia patients receiving haploidentical killer-immunoglobulin-like-receptor-mismatched NK cells, with induction of complete remission was the determinant to begin an increasing number of clinical studies administering NK cells for the treatment of cancer patients. Unfortunately, even though transfused NK cells demonstrated safety, their observed efficacy was poor. In recent years, novel studies have emerged, combining NK cells with other immunotherapeutic agents, such as monoclonal antibodies, which might improve clinical efficacy. Moreover, genetically-modified NK cells aimed at arming NK cells with better efficacy and persistence have appeared as another option. Here, we review novel pre-clinical and clinical studies published in the last five years administering NK cells as a monotherapy and combined with other agents, and we also review chimeric antigen receptor-modified NK cells for the treatment of cancer patients. We then describe studies regarding the role of NK cells as anti-microbial effectors, as lessons that we could learn and apply in immunotherapy applications of NK cells; these studies highlight an important immunoregulatory role performed between T cells and NK cells that should be considered when designing immunotherapeutic strategies. Lastly, we highlight novel strategies that could be combined with NK cell immunotherapy to improve their targeting, activity, and persistence.

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Section: Antibody-based Constructsmentioning

confidence: 99%

Natural Killer Cells in Immunotherapy: Are We Nearly There?

Bachiller

Battram

Perez-Amill

et al. 2020

Cancers

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“…In order to construct a second-generation TriKE capable of both ADCC and NK cell expansion, we modified our previously reported TriKE platform [ 22 ]. A wild-type human IL-15 crosslinker with two modified flanking regions was inserted between two antibody fragments—an n-terminal VHH humanized camelid anti-CD16 fragment [ 23 ] and a c-terminal anti-B7-H3 fragment [ 27 ]—creating cam1615B7H3. Figure 1 A shows a schematic of the B7-H3 TriKE construct.…”

Section: Resultsmentioning

confidence: 99%

“…Single-domain V H H antibodies derived from camelids are known to offer advantages over conventional V L -V H scFv fragments. Thus, we spliced the complementary determining regions (CDRs) from a camelid (llama) anti-CD16 [25] into a universal, humanized, heavy chain scaffold [23,26]. This humanized camelid sequence was used to manufacture cam1615B7H3.…”

Section: Construction Of Cam1615b7h3 Trikesmentioning

confidence: 99%

“…The final product was greater than 90% pure with a molecular weight of about 55 kDa; the predicted molecular weight was 54.58 kDA. As with other TriKE molecules, this TriKE is expected to have a rapid clearance profile due to its size, with EC50 ranges in the order of a couple of hours [23]. Chromatography trace from the second-step purification of cam1615B7H3 on a size exclusion chromatography (SEC) column.…”

Section: Creation and Purification Of A B7-h3 Targeting Trikementioning

confidence: 99%

“…Due to these characteristics, we have designed and described a tri-specific killer engager (TriKE) platform consisting of a single chain variable fragment (scFv) targeting CD16, the most potent activating receptor on NK cells, a scFv targeting a tumor associated antigen, and an IL-15 moiety [18][19][20][21][22]. Recently, we have improved on this platform by adding a single domain antibody against CD16, the result of which is better IL-15 activity and overall function [23]. IL-15 is the most critical homeostatic cytokine for NK cell function [24].…”

Section: Introductionmentioning

confidence: 99%

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NK-Cell-Mediated Targeting of Various Solid Tumors Using a B7-H3 Tri-Specific Killer Engager In Vitro and In Vivo

Vallera

Ferrone

Kodal

et al. 2020

Cancers

Self Cite

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We improved the bispecific antibody platform that primarily engages natural killer (NK) cells to kill cancer cells through antibody-dependent cellular cytotoxicity (ADCC) by adding IL-15 as a crosslinker that expands and self-sustains the effector NK cell population. The overall goal was to target B7-H3, an established marker predominantly expressed on cancer cells and minimally expressed on normal cells, and prove that it could target cancer cells in vitro and inhibit tumor growth in vivo. The tri-specific killer engager (TriKETM) was assembled by DNA shuffling and ligation using DNA encoding a camelid anti-CD16 antibody fragment, a wild-type IL-15 moiety, and an anti-B7-H3 scFv (clone 376.96). The expressed and purified cam1615B7H3 protein was tested for in vitro NK cell activity against a variety of tumors and in vivo against a tagged human MA-148 ovarian cancer cell line grafted in NSG mice. cam1615B7H3 showed specific NK cell expansion, high killing activity across a range of B7-H3+ carcinomas, and the ability to mediate growth inhibition of aggressive ovarian cancer in vivo. cam1615B7H3 TriKE improves NK cell function, expansion, targeted cytotoxicity against various types of B7-H3-positive human cancer cell lines, and delivers an anti-cancer effect in vivo in a solid tumor setting.

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Emerging Antibodies in Cancer Therapy

Sun

2022

Advanced NanoBiomed Research

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Since the first monoclonal antibody, Orthoclone OKT3, was approved in 1986 for the treatment of acute allograft rejection in renal transplant recipients, the number of antibody‐based therapies has increased remarkably, with more than 130 monoclonal antibodies being currently available. In particular, due to their robust binding affinity and high specificity, various monoclonal antibodies have achieved success as complementary antitumor drugs. Moreover, therapeutic potency can be improved by engineering Y‐shaped binding sites of the antibodies to simultaneously recognize two distinct antigens; such molecules are called bispecific antibodies and function as a bridge, with one site binding to a tumor‐specific antigen and the other to immune cells to activate host immune responses. In addition, the development of trispecific antibodies has led to further enhancement of therapeutic specificity and effects. Indeed, a variety of structurally engineered antibodies have proven effective in tumor therapy. This article provides insights into the clinical translation of emerging antibody constructs, firstly describing conventional T cell‐redirecting bispecific antibodies and subsequently discussing the application of bispecific and trispecific antibody constructs based on innate immune cell recruitment. Finally, bispecific antibodies against mutant p53 and KRAS that have been actively studied in cancer treatment are introduced.

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Potent Cytolytic Activity and Specific IL15 Delivery in a Second-Generation Trispecific Killer Engager

Cited by 48 publications

References 43 publications

Natural Killer Cells in Immunotherapy: Are We Nearly There?

Natural Killer Cells in Immunotherapy: Are We Nearly There?

NK-Cell-Mediated Targeting of Various Solid Tumors Using a B7-H3 Tri-Specific Killer Engager In Vitro and In Vivo

Emerging Antibodies in Cancer Therapy

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