Development of Novel sgp130-Fc Bioconjugates for TBI

摘要 患有中度至重度创伤性脑损伤（TBI）的个体处于多个长期 并发症和神经恢复差。尽管对急性继发性损伤级联的认识有所增加， 对伴随继发性疾病的慢性病理学的潜在机制知之甚少， 影响TBI结果。因此，在TBI的慢性康复阶段的治疗方面存在差距 支持神经恢复并减轻继发性疾病的风险。我们发表的临床研究 表明急性脑脊液（CSF）IL-6水平与严重TBI后的结果相关， 这些第一周的时间IL-6谱确定了潜在的亚急性和慢性外周炎症 包括IL-6在内的与长期功能结局相关的标志物。我们发表的研究表明， 损伤后前3个月内血清sIL-6 R水平与总体认知能力较差相关 在损伤后6个月和12个月进行评估，但sgp 130/sIL-6 R的比率较高与更好的认知功能相关。 测试性能表明sgp 130对sIL-6 R相关的“反式信号传导”的潜在保护作用。 抑郁症也与sIL-6 R水平有关，我们的临床数据显示sIL-6 R对总体抑郁症的调节作用。 结果，其中高sIL-6 R信号负荷有利于有害的IL-6“反式信号”环境， 我们提出，促进CNS损伤，而低sIL-6 R负荷有利于有益的经典信号，我们提出， 支持神经恢复这些临床数据表明，sIL-6 R是急性胰腺炎后的一个可调节的靶点。 在TBI恢复的康复阶段，sgp 130可能是改善结果的可行治疗。 这些临床研究结果得到了使用受控皮质撞击损伤的体内研究的补充 (CCI)小鼠和大鼠中的TBI模型显示sgp 130-Fc生物缀合物可以减少认知缺陷、CNS促神经生长、 炎症信号传导和与CCI相关的组织学损伤。然而，各种突变和 sgp 130-Fc的缀合物，包括抗转铁蛋白受体（抗mTfR Oligo）sgp 130-Fc生物缀合物，可以 促进血脑屏障（BBB）穿透并减少TBI后CNS sIL-6 R反式信号传导。因此，该SBIR 将专注于体外概念验证研究，以开发和评估新型sgp 130-Fc生物缀合物，其中IgG 1 mAb作为同种型对照（I期），开发新型抗TfR寡核苷酸/sgp 130-Fc生物缀合物，并评估其 使用TBI的CCI小鼠模型的体内作用（II期）。我们将通过以下方式选择主要生物偶联物候选物： 在严重CCI后的雄性和雌性小鼠中测试治疗后的1°和2°终点的功效。这些 包括PK测定、小胶质细胞活化/摄取、sIL-6 R反式信号传导阻断和治疗功效。 将通过检查促炎和CNS的血清/CNS负荷降低来评估治疗功效。 生物标志物，脑组织保护，细胞免疫正常化，神经行为改善 评估。这项工作将支持未来的研究开发可行的人sgp 130-Fc生物缀合物， 中重度TBI后的临床、康复阶段治疗。

ABSTRACT Individuals with moderate-to-severe traumatic brain injuries (TBI) are at high risk for multiple long-term complications and poor neuro-recovery. Despite increased knowledge about acute secondary injury cascades, far less is known about mechanisms underlying the chronic pathology that accompany secondary conditions and influence TBI outcome. Thus, there is a gap in therapeutics for the chronic, rehabilitation phases of TBI that support neurorecovery and mitigate risk for secondary conditions. Our published clinical research suggests that acute cerebrospinal fluid (CSF) IL-6 levels are associated with outcome after severe TBI, and these temporal IL-6 profiles over the first week identified potential sub-acute and chronic peripheral inflammatory markers, including IL-6, that associate with long-term functional outcome. Our published work shows that higher serum sIL-6R during the first 3 months post-injury are associated with worse overall cognitive performance assessed 6- and 12- months post-injury, yet higher ratios of sgp130/sIL-6R are associated with better cognitive testing performance indicating a potential protective effect of sgp130 against sIL-6R associated “trans-signaling”. Depression is also linked to sIL-6R levels, and our clinical data show a moderating effect of sIL-6R on global outcomes wherein high sIL-6R signaling loads favor a detrimental IL-6 “trans-signaling” environment that we propose facilitates CNS damage, while low sIL-6R loads favor beneficial classical signaling that we propose supports neurorecovery. These clinical data suggest that sIL-6R is a modifiable target in the post-acute rehabilitation phase of TBI recovery for which sgp130 may be a viable treatment that improves outcome. These clinical research findings are complemented by in vivo studies using the controlled cortical impact injury (CCI) model of TBI in mice and rats showing sgp130-Fc bioconjugates can reduce cognitive deficits, CNS pro- inflammatory signaling, and histological damage associated with CCI. However, various mutations and conjugations of sgp130-Fc, including anti-transferrin receptor (anti-mTfR Oligo) sgp130-Fc bioconjugates, may facilitate blood brain barrier (BBB) penetration and reduce CNS sIL-6R trans-signaling after TBI. Thus, this SBIR will focus on proof-of-concept in vitro studies to develop and assess novel sgp130-Fc bioconjugates, with IgG1 mAbs as isotype controls (Phase I), and develop novel anti-TfR Oligo/sgp130-Fc bioconjugates and assess their effects in vivo using the CCI mouse model of TBI (Phase II). We will select lead bioconjugate candidates by testing 1° and 2° endpoints for efficacy following treatment in male and female mice after severe CCI. These include PK assays, microglial activation/uptake, sIL-6R trans-signaling blockade, and treatment efficacy. Treatment efficacy will be assessed by examining reduced serum/CNS load of pro-inflammatory and CNS biomarkers, brain tissue sparing, normalization of cellular immunity, and improvements in neurobehavioral assessments. This work will support future studies developing viable human sgp130-Fc bioconjugates for clinical, rehabilitation phase treatment after moderate to severe TBI.