Both PD1/PD-L1 and CD47 blockades have demonstrated limited activity in most subtypes of NHL save NK/T-cell lymphoma. The hemotoxicity with anti-CD47 agents in the clinic has been speculated to account for their limitations. Herein we describe a first-in-class and rationally designed bispecific antibody (BsAb), HX009, targeting PD1 and CD47 but with weakened CD47 binding, which selectively hones the BsAb for tumor microenvironment through PD1 interaction, potentially reducing toxicity. In vitro characterization confirmed: (1) Both receptor binding/ligand blockade, with lowered CD47 affinity; (2) functional PD1/CD47 blockades by reporter assays; (3) T-cell activation in Staphylococcal-enterotoxin-B-pretreated PBMC and mixed-lymphocyte-reaction. In vivo modeling demonstrated antitumor activity in Raji-B and Karpass-229-T xenograft lymphomas. In the humanized mouse syngeneic A20 B-lymphoma (huCD47-A20) HuGEMM model, which has quadruple knocked-in hPD1xhPD-L1xhCD47xhSIRPα genes and an intact autologous immune-system, a contribution of effect is demonstrated for each targeted biologic (HX008 targeting PD1 and SIRPα-Fc targeting CD47), which is clearly augmented by the dual targeting with HX009. Lastly, the expression of the immune-checkpoints PD-L1/L2 and CD47 seemed co-regulated among a panel of lymphoma-derived-xenografts, where HX009 maybe more effective in those with upregulated CD47. Our data warrants HX009’s further clinical development for treating NHLs.

Acute myeloid leukemia (AML) is an aggressive leukemia of myeloid lineage with different subtypes (global annual mortality ~150,000). The first-line treatment is usually induction chemotherapies, followed by further chemo-/radiation therapies or stem cell transplant, etc. Immune checkpoint inhibitor (ICIs), e.g. PD1 blocker, although a great success for treating many solid tumors, has yet to show convincing activity in AML. CD47, a “don’t eat me” receptor frequently found on the surface of tumor cells, including hematological malignances such as AML, prevents phagocytosis by interacting to its ligand, SIRPα, on the surface of macrophages. CD47 is also now shown as a key immune checkpoint for both innate and adoptive immunity (Liu et al., 2018). CD47 targeting, actively being tested for MSD and AML in clinics, encounters challenges for the reported toxicities (e.g. anemia/thrombocytopenia) as well as inadequate efficacy (Feng et al., 2019). Majority of investigational CD47 targeting agents in the clinics are combination therapies with other modality in order to increase the likelihood of success. To this end, it is prudent to develop agents with dual targeting agents, particularly for both the innate and adoptive immune checkpoints. We describe a novel specially designed/constructed bi-specific antibody (BsAb), HX009, targeting CD47 and PD1 for the treatment of cancers. We tested HX009 in several preclinical AML-PDXs (patient-derived xenograft models) via systemic engraftment: AM8096 (CD47lo, H-score of 55 by IHC), AM7577 (CD47hi, H-score of 237)(An et al., 2017) and AM5512 (CD47hi, H-score of 198). All three PDXs responded to 10mg/kg twice weekly dose of HX009 (IP) as measured either by leukemic burden in peripheral blood at different timepoints, and in both spleen and bone marrow at the termination, or by survival. Since there is no T-cell function in xenograft models, all anti-leukemic activities observed should be attributed to the CD47 blockade, not PD1 blockade. AM7577 model responded significantly better than AM8096, likely due to the significantly higher expression of CD47 and also better than Ara-C standard of care (SOC) treatment at 3mg/kg. In contrast, AM8096 responded even less to HX009 than Ara-C. In contrast, in the subcutaneous transplanted AML cell line-derived model, MV4-11, there is little anti-tumor activity observed, although there is significant expression of CD47 on the tumor cells. In conclusion, our data seems to suggest that HX009 could be a candidate immunotherapy for CD47hi AML, with CD47 expression being a positive predictive biomarker, warranting further evaluation. Citation Format: Xiaoyu An, Jinping Liu, Hang Ke, Lingxin Xiong, Xiaolong Tu, Lei Zhang, Faming Zhang, Henry Li. HX009, a first-in-class PD1xCD47 BsAb, demonstrated anti-AML activity in PDX models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2980.

In the process of sailing, the frictional resistance of underwater objects must be reduced, which is of great significance for improving speed, range and saving energy. Based on the inspiration of the shark breathing process, this paper simplified the water spray process from the external bronchial slit of the shark breathing to the cross-flow lateral jet process. By establishing a numerical model, the influence mechanism of the jet on the flow resistance was studied, and by analyzing different jet velocities, the change of viscous resistance reveals the influence of jet velocity on viscous resistance. The results show that the side jet can reduce the boundary layer on the surface of the object, thereby having a drag reduction effect, and the drag reduction rate can be about 10%. In addition, as the jet velocity increases, the drag reduction effect is enhanced, and at the back of the jet port the vortex is formed, the outlet pressure is reduced, and the drag reduction effect is strengthened. At the same time, the study found that the arc-shaped jet port is more conducive to the reduction of resistance.

No abstract

OX40, tumor necrosis factor receptor superfamily member 4, is an immune co-stimulatory receptor, together with its ligand OX40L, forming OX40-OX40L trimer-trimer complex between the surface of T-cells (OX40) and APCs/NKs (OX40L). OX40 expresses constitutively on Treg as well as on conventional Teff, particularly within TME. OX40/OX40L interactions directly triggers conventional T cell activation via NF-kB1 pathway and inhibits bcl-x and survivin to prevent apoptosis; and it also downregulates FoxP3/CTLA4, thus Treg function. OX40 agonistic antibodies have also been shown to activate tumor immunity and increase anti-tumor activity in preclinical models. Here we report that we identified/constructed a novel OX40-agonistic humanized IgG1 monoclonal antibody (mAb), HX011, with high affinity through hybridoma technology. HX011 was subsequently confirmed of the similar binding properties and functions as the original mouse mAb clone, 9C12: 1) binding to human OX40 (EC50 ~0.012 nM; 2) similar function binding in the reporter assay (EC50 ~6.6 nM); but 3) has little blockade of the OX40L binding to OX40. In another word, HX011 is similar to BMS-986178 for the first two assays, but contrastingly different for the third assay. Non-competing with OX40L may actually maximize T-cell activation, a potential advantage over a competing antibody. In vivo pharmacology modeling of HX011 in the humanized syngeneic CRC model, MC38, where syngeneic C57BL/6 mouse mice were knocked in with huOX40 (“HuGEMM-OX40”), demonstrated strong anti-tumor activity (TGI ~100% at 1 mg/kg, twice weekly). HX011 is being further investigated as a potential new immunotherapy for the treatment of cancers. Citation Format: Hang Ke, Faming Zhang, Jialin Li, Feiyu Peng, Cen Chen, Henry Qixiang Li. Discovery of HX011, a novel OX40 mAb, for potential cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6346.