Immunogenicity, Inflammation, and Lipid Accumulation in Cynomolgus Monkeys Infused with a Lipidated Tetranectin-ApoA-I Fusion Protein

Abstract

High density lipoprotein (HDL)-targeted therapies, which promote cholesterol efflux from cells, are currently in development for reducing cardiovascular events in acute coronary syndrome. Human apolipoprotein A-I (apoA-I), the major HDL protein, was fused to the trimerization domain of tetranectin (TN) and complexed with phospholipids to generate a HDL mimetic (lipidated TN-ApoA-I) with reduced renal clearance and enhanced efficacy. Cynomolgus monkeys received 24-h intravenous infusions of control, 100 mg/kg or 400 mg/kg lipidated TN-ApoA-I every 4 days for 3 weeks, followed by a 6-week recovery period. After multiple infusions of lipidated TN-ApoA-I, clinical condition deteriorated and was accompanied by changes indicative of a progressive inflammatory response; increased levels of cytokines, C-reactive protein and vascular/perivascular infiltrates in multiple tissues. Rapid formation of antidrug antibodies occurred in all animals receiving lipidated TN-ApoA-I. Enhanced drug clearance corresponding to a relative lack of high molecular weight immune complexes in blood, suggestive of preferred removal/clearance, was observed in some animals. Expected dose-dependent increases in serum lipids were accompanied by vacuolated monocytes/macrophages in multiple organs, which in the glomeruli were shown to be CD68-positive, contain lipid and co-localized with granular IgG deposits. Lipid accumulation may have been a direct result of a high drug load, possibly enhanced by immune complex formation, inflammation, and altered lipid metabolism. Noteworthy was the inter- individual inconsistency in the severity of clinical and histopathologic findings, drug clearance and inflammatory markers. In conclusion, multiple infusions of lipidated TN-ApoA-I resulted in high immunogenicity, lipid accumulation and were not well tolerated in nonhuman primates.

The enhanced immunopharmacology of VIB4920, a novel Tn3 fusion protein and CD40L antagonist, and assessment of its safety profile in cynomolgus monkeys

“…In Study 1, three animals receiving higher VIB4920 doses (150 mg·kg −1 i.v. [ n = 2] and 150 mg·kg −1 s.c. [ n = 1] weekly) had minimal Kupffer cell hypertrophy, which is commonly seen during high‐dose protein administration (Maclaren, Levin, Lowman, & Trikha, ; Regenass‐Lechner et al, ).…”

“…VIB4920-treated animals and in one vehicle-treated animal. (Maclaren, Levin, Lowman, & Trikha, 2017;Regenass-Lechner et al, 2016). (Iverson et al, 2018).…”

Biomarkers for nonclinical infusion reactions in marketed biotherapeutics and considerations for study design

“…1 ). Large immune complexes can be cleared by the mononuclear phagocytic system, small immune complexes can recirculate and be cleared or become mid-sized immune complexes, which have been shown to result in tissue deposition ∗∗2 , 3 , ∗∗4 .

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“…Complement activation appears to be related to the size of the immune complex. Insoluble, large immune complexes of ADA bound to a biotherapeutic are cleared rapidly by the liver and spleen via the mononuclear phagocytic system ∗∗2 , 3 , ∗∗4 . Conversely, smaller (dimer) soluble immune complexes are present in circulation longer, and can facilitate receptor recycling of the biotherapeutic, which can increase the size of the IC and ADA titer, or may result in class switching from IgM to IgG-ADA, which can produce a more severe IR and subsequent tissue deposition of the immune complex [3] .…”

“…Minimize freeze/thaw cycles Decreased 2-4 h after IR C3a serum or plasma 0.5–1.8 Increased C5a Increased Bb Increased Sc5b-9 Increased Cytokines ∗∗2 , ∗∗5 , 10 , 12 , 19 , ∗50 , ∗∗51 Multi-plexed assay (cytokines most commonly observed listed) IL-6, TNF-α, IFNγ, IL1-b, IL-2 serum or plasma 0.5–1.0 Process and freeze immediately. Minimize freeze/thaw cycles Increased 2-4 h after IR Coagulation ∗∗2 , ∗∗4 , ∗∗5 APTT plasma 1–1.8 Samples taken by different routes or in the presence of anesthetics may impact biomarker modulation [63] Increased 2-4 h after IR PT Increased Fibrinogen c , e Decreased or increased D-dimer Increased Hematology ∗∗2 , ∗∗5 , ∗52 Red blood cell mass (RBCs, Hgb, Hct) plasma 0.5–1 Samples taken by different routes or in the presence of anesthetics may impact biomarker modulation [63] d Increased or Decreased 2-4 h after IR Platelets d , e Decreased Neutrophils d Decreased or Increased Lymphocytes d Unchanged or Increased ...…”