US-12625147-B2 - Molecular tools to visualize and target the cardiac conduction system (CCS)

US12625147B2US 12625147 B2US12625147 B2US 12625147B2US-12625147-B2

Abstract

Antibodies which bind selectively to cardiac conduction system (CCS) cells, imaging and/or diagnostic reagents and compositions visualizing the CCS cells and therapeutic products and compositions comprising one or more of the antibodies. Methods for delivering therapeutic agents to the CCS cells. The disclosure further provides methods for visualizing the CCS cells in vivo in real time, including in a subject undergoing a cardiothoracic surgery or other cardiac intervention. Compositions and methods for isolation, purification, analyses and/or transplantation of the CCS cells, including pluripotent stem cell (hiPSC)-derived or human embryonic stem cell (hESC)-derived CCS cells.

Inventors

Sean M. Wu
William R. Goodyer
Benjamin Beyersdorf
Nynke Van Den Berg
Eben Rosenthal

Assignees

THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITY

Dates

Publication Date: 20260512
Application Date: 20200707

Claims (7)

1 . A method for visualizing in real time cardiac conduction system (CCS) in a subject undergoing cardiothoracic surgery or catheter procedure, the method comprising: a) administering topically, intravenously or intracoronary or by preoperative infusion to the subject a composition comprising one or more excipients and an antibody conjugated directly to a detection agent which comprises a fluorophore, wherein the antibody has a specific binding affinity to Neurotrimin, Neuroplastin or Contactin 2 displayed on a CCS cell, and wherein subsequent to the administration, the antibody conjugate binds to the CCS cell; and b) illuminating the subject's heart and thereby visualizing the bound antibody conjugate in the subject while the subject is undergoing the cardiothoracic surgery or catheter procedure.
2 . The method of claim 1 , wherein the method further comprises capturing images of the CCS in real time with camera.
3 . The method of claim 1 , wherein the composition comprises from 0.1 wt % to 99.9 wt % of the antibody conjugate and from 0.1 wt % to 99.9 wt % of the one or more excipients.
4 . The method of claim 1 , wherein the fluorophore is a biocompatible near-infrared (NIR) fluorophore.
5 . The method of claim 1 , wherein the subject's heart is illuminated with infrared light.
6 . The method of claim 1 , wherein the subject's heart is illuminated with a scope placed into the subject.
7 . The method of claim 1 , wherein the antibody is conjugated covalently to the detection agent.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a U.S. national stage application of PCT application PCT/US2020/040965 with the international filing date of Jul. 7, 2020 which claims the benefit of priority from U.S. provisional application 62/871,551 filed on Jul. 8, 2019 and U.S. provisional application 62/950,428 filed on Dec. 19, 2019, the combined disclosure of all these applications is herein incorporated by reference. STATEMENT OF GOVERNMENT INTEREST This invention was made with Government support under contract LM012179 awarded by the National Institutes of Health. The Government has certain rights in the invention. TECHNICAL FIELD The invention relates to imaging reagents for the cardiac conduction system (CCS) cells and compositions and methods for delivering therapeutic agents to the CCS cells. The invention further relates to compositions and methods visualizing the CCS cells in vivo in real time, including in a subject undergoing a cardiothoracic surgery or other cardiac intervention. In further aspect, the invention relates to compositions and methods for isolation, purification, analyses and/or transplantation of the CCS cells, including pluripotent stem cell (hiPSC)-derived or human embryonic stem cell (hESC)-derived CCS cells. BACKGROUND The cardiac conduction system (CCS) is made up of specialized heart cells that establish the rhythmic beating of the heart through coordinated contraction of its chambers. The cardiac conduction system (CCS) is comprised of distinct components including the sinoatrial node (SAN), atrioventricular node (AVN), His bundle (HIS), bundle branches (BB) and Purkinje fibers (PF). The CCS is essential for the formation and normal function of the heart and disturbance to the CCS can result in severe clinical manifestations including arrhythmias, decreased cardiac output and even sudden death. Despite an essential role for the CCS in heart development and function, the CCS has remained difficult to study due to inherent obstacles including small cell numbers, large cell type heterogeneity, complex anatomy and difficulty in isolation. Each component of the CCS consists of unique cardiac cell types with their own physiologic and electrochemical properties.4 Further, the CCS components each have significant intracomponent cell type heterogeneity.5 The most clinically relevant example of this heterogeneity has been the discovery of “transitional cells”, that exist in each CCS component and provide a cellular bridge to the surrounding working myocardium.6-10 They are hypothesized to play a role in facilitating the spread of depolarization, providing a high resistance barrier as well as amplifying the current before passing it on to the surrounding myocardium.9 Transitional cells have been implicated in several important clinical disorders including sinus node dysfunction, heart block and even ventricular fibrillation. However, they remain poorly understood due to challenges in their identification, isolation and, ultimately, molecular characterization. Currently, the limited number of distinct molecular markers that are known for the different CCS cell types present a number of challenges for additional investigation into their specification, patterning and function. While individual factors have been examined thus far, these studies have been performed at tissue but not at single-cell resolution.12-14 Specific hurdles to better understanding the molecular signature of the CCS cells have included: 1) low total number of conduction cells in the heart; 2) complex three-dimensional anatomy of the CCS; 3) inability to isolate these cells from the surrounding working myocardium; and 4) the aforementioned significant inter- and intra-component cell type heterogeneity.1 While prior studies have assessed gene expression within individual components of the CCS using sophisticated techniques such as microdissection, laser capture or even fluorescence-activated cell sorting12-21, these analyses are unable to discern cell type heterogeneity due to bulk tissue analysis and/or a reliance on transgenic fluorescent reporter models. The CCS is essential for the normal formation and function of the heart, and injury to the CCS can result in heart block, arrhythmias, decreased cardiac output and even sudden death. Congenital heart disease remains the most common birth defect, affecting roughly 1 in 100 infants, with a significant fraction requiring corrective heart surgery. Iatrogenic surgical damage to the CCS remains a significant complication in both congenital heart disease (CHD) and adult cardiac surgeries. This is due in part to an inability to visualize, and thus, avoid damaging the surrounding CCS by inadvertent incision or suture placement. Accidental intraoperative injury to the cardiac conduction system (CCS) complicates roughly 1-3% of all congenital heart disease surgeries and an even higher percentage of the ˜106,000 adult heart valve surgeries performed in the Unit