Development of Novel sgp130-Fc Bioconjugates for TBI

摘要 中至重度创伤性脑损伤(TBI)患者长期多发的风险很高。 并发症和神经恢复差。尽管人们对急性继发性损伤级联反应的了解有所增加， 对伴随继发性疾病和慢性疾病的慢性病理的潜在机制知之甚少 影响颅脑损伤的结果。因此，脑外伤的慢性康复阶段的治疗方法存在缺口。 支持神经康复并减轻继发性疾病的风险。我们已发表的临床研究 提示急性脑脊液IL-6水平与重型颅脑损伤后的预后相关。 第一周的暂时性IL-6分析确定了潜在的亚急性和慢性外周炎症 包括IL-6在内的与长期功能预后相关的标志物。我们发表的研究表明，更高的 损伤后前3个月的血清sIL-6R与整体认知能力较差有关 在损伤后6个月和12个月进行评估，但sgp130/sIL-6R的较高比率与较好的认知能力相关 测试性能表明，sgp130对sIL-6R相关的“转导信号”具有潜在的保护作用。 抑郁症也与sIL-6R水平有关，我们的临床数据显示sIL-6R对全球 结果高sIL-6R信号负荷有利于有害的IL-6“转导信号”环境 Proposal促进中枢神经系统损伤，而低sIL-6R负荷有利于我们提出的有益的经典信号转导 支持神经康复。这些临床数据表明，sIL-6R是急性心肌梗死后的一个可修改的靶点。 创伤性脑损伤恢复的康复阶段，sgp130可能是一种可行的改善预后的治疗方法。 这些临床研究结果得到了使用受控皮质撞击损伤的活体研究的补充。 显示sgp130-Fc生物偶联物可减少认知功能障碍的大鼠和小鼠脑外伤模型，中枢神经系统亲 与CCI相关的炎症信号和组织损伤。然而，各种突变和 Sgp130-Fc的结合物，包括抗转铁蛋白受体(抗mTfR寡核苷酸)sgp130-Fc生物结合物，可以 促进血脑屏障穿透，减少中枢sIL-6R反式信号转导。因此，该SBIR 将专注于概念验证的体外研究，以开发和评估新型的sgp130-Fc生物结合物，与IgG1 单抗作为同型对照(I期)，开发新型抗转铁蛋白受体寡核苷酸/sgp130-Fc生物偶联物并评价其 使用CCI小鼠模型的活体效应(第二阶段)。我们将通过以下方式选择铅生物结合物候选者 在雄性和雌性小鼠严重CCI后，测试1°和2°终点对治疗后的疗效。这些 包括PK检测、小胶质细胞活化/摄取、sIL-6R转导信号阻断和治疗效果。 治疗效果将通过检测促炎和中枢神经系统减少的血清/中枢神经系统负荷来评估 生物标记物、脑组织保护、细胞免疫正常化和神经行为改善 评估。这项工作将支持未来开发可行的人sgp130-Fc生物结合物的研究 中重度颅脑损伤后的临床、康复期治疗。

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.