Compositions and methods for RNA delivery to cells

Abstract

Provided are molecular hooks that are capable of tethering a ribonucleic acid (RNA) of interest to a high-density lipoprotein (HDL) molecule. Also provided are methods of RNA delivery to cells and organisms using the molecular hooks of the present disclosure. Such methods provide several advantages including the ability to deliver the RNA of interest to tissues behind the blood-brain barrier, and not requiring extreme cold temperatures for storage or transport.

Description

Atty. Dkt. No.: 139886-1010 COMPOSITIONS AND METHODS FOR RNA DELIVERY TO CELLS CROSS-REFERENCE TO RELATED PATENT APPLICATION

[0001] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application No.63 / 635,707, filed April 18, 2024, the entire contents of which are incorporated herein by reference. BACKGROUND

[0002] All RNA-based therapies, including but not limited to messenger RNA (mRNA), microRNA (miRNA), small interfering RNA (siRNA), and antisense RNA molecules—as well as RNA-based vaccines and related therapeutics—depend on effective intracellular delivery mechanisms to achieve their intended biological functions. Current RNA delivery platforms, such as lipid nanoparticle (LNP) systems, suffer from several inherent limitations.

[0003] Firstly, LNPs are synthetic chemical constructs that may demonstrate cumulative toxicity upon repeated or long-term use, thereby limiting their applicability as delivery vehicles in chronic or multi-dose treatment regimens.

[0004] Secondly, in order to formulate mRNA as a stable therapeutic product suitable for commercial storage and distribution (e.g., in vials or ampoules), it must undergo stabilization. This typically necessitates ultra-low temperature conditions for storage and transport, frequently at –50°C or lower, to preserve structural integrity and biological activity.

[0005] Furthermore, the capacity of currently available LNP systems to cross the blood-brain barrier (BBB) remains controversial, with conflicting evidence in the literature.

[0006] Accordingly, in order to unlock the full therapeutic potential of RNA-based technologies, there exists a pressing and unmet need for the development of enhanced RNA stabilization strategies; and the creation of novel, biocompatible, and non-toxic delivery systems capable of achieving efficient intracellular transport without compromising safety. SUMMARY OF THE DISCLOSURE

[0007] In one aspect, the present disclosure is directed to an in vivo method for delivering ribonucleic acid (RNA) to cells of a subject in need thereof comprising administering to the subject (i) an RNA-HDL complex wherein the RNA is tethered to the exterior of a high- density lipoprotein (HDL), or (ii) an RNA-molecular hook complex wherein the molecular hook is capable of tethering to the exterior of any HDL molecule. 1 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0008] In another aspect, the present disclosure is directed to a method for delivering ribonucleic acid (RNA) to a cell, the method comprising contacting the cell with an RNA– HDL complex, wherein the RNA is tethered to the outer surface of a high-density lipoprotein (HDL) particle. Optionally, in certain embodiments, the RNA is bound to the lipid outer layer of the HDL via a molecular hook. In various embodiments, said contacting may occur under in vitro or in vivo conditions.

[0009] In some embodiments, the cells of the subject are behind the blood brain barrier of the subject. In some embodiments, the cells of the subject are in the brain or the spine of the subject. In a further embodiment, the cells of the subject are behind the blood brain barrier of the subject and are located in the brain or the spine of the subject.

[0010] In some embodiments, the molecular hook comprises: a first antigen binding domain that specifically binds to an exterior facing protein of the HDL; and a second antigen binding domain that specifically binds to the ribonucleic acid (RNA).

[0011] In some embodiments, the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C-II (Apo C-II).

[0012] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or accession No. NITE BP-02443.

[0013] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 .

[0014] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO: 68. 2 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0015] In some embodiments, the first antigen binding domain specifically binds ApoA-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the EPR2913 monoclonal antibody, or amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

[0016] In some embodiments, the first antigen binding domain specifically binds Apo E and comprises an amino acid sequence selected from: (i) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of MAB41445 monoclonal antibody; (ii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 42; (iii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 44; (iv) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 46; (v) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47 and CDR-H1, CDR-H2 and CDR- 3 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 48; (vi) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 50; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 41 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 42; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 44; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 45 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 46; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 47 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 48; or (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 49 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 50.

[0017] In some embodiments, the first antigen binding domain specifically binds Apo C-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the 3E4 monoclonal antibody.

[0018] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises HuR RNA binding protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 1.

[0019] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 4 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 .

[0020] In some embodiments, the first antigen binding domain and or the second antigen binding domain comprises an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody.

[0021] In some embodiments, the first antigen binding domain is from a recombinant antibody. In some embodiments, the second antigen binding domain is from a recombinant antibody.

[0022] In some embodiments, the molecular hook comprises a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker.

[0023] In one aspect the molecular hooks further comprise a detectable label.

[0024] In another aspect, this disclosure provides DNA polynucleotides encoding the molecular hooks as described herein, optionally detectably labeled. Vectors and host cells comprising the polynucleotides are further provided herein. The cells can be prokaryotic or eukaryotic.

[0025] In some embodiments, the RNA comprises mRNA, and optionally wherein the mRNA encodes a therapeutic protein; and optionally wherein the mRNA encodes a vaccine.

[0026] In some embodiments, the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain of the anti-APOA-I binding antibody and the HuR RNA binding protein are linked together by a flexible linker selected from SEQ ID NOs: 76-79

[0027] In some embodiments, the molecular hook comprises SEQ ID NO: 75 and SEQ ID NO: 67.

[0028] Another aspect of the disclosure is directed to a molecular hook comprising: a first antigen binding domain that specifically binds to an exterior facing protein of a high density lipoprotein (HDL); and a second antigen binding domain that specifically binds to a ribonucleic acid (RNA). 5 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0029] In some embodiments, the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C (Apo C).

[0030] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 comprising amino acid sequences that are respectively same as amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or accession No. NITE BP-02443. In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 .

[0031] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO: 68.

[0032] In some embodiments, the first antigen binding domain specifically binds ApoA-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the EPR2913 monoclonal antibody, or amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

[0033] In some embodiments, the first antigen binding domain specifically binds Apo E and comprises an amino acid sequence selected from: 6 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 (i) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of MAB41445 monoclonal antibody; (ii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 42; (iii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 44; (iv) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 46; (v) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 48; (vi) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49 and CDR-H1, CDR-H2 and CDR- H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR- H3 of SEQ ID NO: 50; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 41 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 42; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 44; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 45 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 46; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 47 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 48; or 7 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 49 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 50.

[0034] In some embodiments, the first antigen binding domain specifically binds Apo C and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the 3E4 monoclonal antibody.

[0035] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises HuR RNA binding protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 1.

[0036] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

[0037] In some embodiments, the first antigen binding domain and or the second antigen binding domain comprises an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody.

[0038] In some embodiments, the molecular hook comprises a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker.

[0039] In some embodiments, the RNA is mRNA; optionally wherein the mRNA encodes a therapeutic protein or a vaccine.

[0040] In some embodiments, the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain of the anti-APOA-I 8 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 binding antibody and the HuR RNA binding protein are linked together by a flexible linker selected from SEQ ID NOs: 76-79

[0041] In some embodiments, the molecular hook comprises SEQ ID NO: 75 and SEQ ID NO: 67.

[0042] In one aspect the molecular hooks further comprise a detectable label. BRIEF DESCRIPTION OF THE FIGURES

[0043] FIGS.1A-1D. (A) Structure of an HDL molecule. HDL molecules are nano sized particles that circulate in the bloodstream. HDL nanoparticles can cross the blood-brain barrier. (B) Schematic of a non-limiting molecular hook example that binds to both ApoA-I and RNA. In this embodiment, the ApoA-I binding part comprises an anti-ApoA-I antibody, or an antigen binding fragment thereof, and the RNA-binding site comprises a HurRNP molecule that can bind to any RNA molecule. (C) Schematic of a non-limiting molecular hook example that binds to both ApoA-I and RNA. In this embodiment, the ApoA-I binding part comprises an anti-ApoA-I antibody, or an antigen binding fragment thereof, and the RNA-binding site comprises an RNA antibody, or an antigen binding fragment thereof. (D) Schematic of a non-limiting molecular hook example that binds to both Apo CII and RNA. In this embodiment, the Apo CII binding part comprises an anti-Apo CII antibody, or an antigen binding fragment thereof, and the RNA-binding site comprises an RNA antibody, or an antigen binding fragment thereof.

[0044] FIG.2. Molecular hooks successfully introduce GFP-mRNA into HEK cells in cell culture by binding to the HDL molecules in human serum used in the experiment. By 96 hours, about 10 percent of the cells showed GFP positivity, which is comparable to other RNA delivery methods used in vivo. PEI: Polyethylenimine (positive control).

[0045] FIGS.3A-3D. Internalization of dye-conjugated antibodies. x-axis: fluorescence intensity, y-axis: event count. Shift in fluorescence indicates internalization of the dye- conjugated compound. (A) Internalization of conjugated ApoA-I antibody versus internalization of an isotype control antibody. The conjugated ApoA-I antibody was internalized in over 71% of the cells. (B) Internalization of conjugated ApoA-I antibody as compared to an untreated (no antibody) control, showing 97% signal in cells treated with the conjugated ApoA-I antibody. (C) Time course of conjugated ApoA-I antibody internalization over 24, 48 or 96 hours. Durable internalization was achieved by 96 hours. 9 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 The rightmost graph (48h + PEI) shows the result of a positive control experiment. In this setup, PEI (polyethylenimine)—a compound known to disrupt cell membrane integrity and to be toxic to virtually all cell types, including both healthy and cancerous cells—was added to the culture medium. Under these conditions, both the control antibody (anti-HEL) and conjugated ApoA-I antibody were internalized in a similar fashion. (D) Quantification of internalization shown in FIG.3C.

[0046] FIGS.4A-4B. Testing for endosome trapping. In these experiments, if the internalized antibody is in the endosomes (trapped in the endosomes) there is an increase in red fluorescence due to the pH change. (A) 20 minutes and (B) 93 hours after conjugated ApoA-I antibody and isotype control antibody treatment cells were imaged in phase contrast and red fluorescence channels. In either time point, there were no endosomal trapping. Same cells were used in FACS experiments to show that the conjugated ApoA-I antibody was indeed internalized into the cell.

[0047] FIGS.5A-5C. Non-limiting molecular hook configurations. (A) A diabody linked to an Fc-Fab. (B) A whole antibody linked to an scFv. (C) A Whole antibody linked to a protein molecule. Any possible combination of the individual antigen binding domains (e.g., a whole antibody, a diabody, a Fab, an scFv, an Fc-Fab, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2 etc.) is possible and contemplated in this application, as described herein. DETAILED DESCRIPTION Definitions

[0048] As it would be understood, the section or subsection headings as used herein is for organizational purposes only and are not to be construed as limiting and / or separating the subject matter described.

[0049] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods, devices, and materials are now described. All technical and patent publications cited herein are incorporated herein by reference in their entirety. Nothing herein is to be construed as an admission that the disclosure is not entitled to antedate such disclosure by virtue of prior disclosure. 10 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0050] The practice of the present disclosure will employ, unless otherwise indicated, conventional techniques of tissue culture, immunology, molecular biology, microbiology, cell biology and recombinant DNA, which are within the skill of the art. See, e.g., Sambrook and Russell eds. (2001) Molecular Cloning: A Laboratory Manual, 3rd edition; the series Ausubel et al. eds. (2007) Current Protocols in Molecular Biology; the series Methods in Enzymology (Academic Press, Inc., N.Y.); MacPherson et al. (1991) PCR 1: A Practical Approach (IRL Press at Oxford University Press); MacPherson et al. (1995) PCR 2: A Practical Approach; Harlow and Lane eds. (1999) Antibodies, A Laboratory Manual; Freshney (2005) Culture of Animal Cells: A Manual of Basic Technique, 5th edition; Gait ed. (1984) Oligonucleotide Synthesis; U.S. Patent No.4,683,195; Hames and Higgins eds. (1984) Nucleic Acid Hybridization; Anderson (1999) Nucleic Acid Hybridization; Hames and Higgins eds. (1984) Transcription and Translation; Immobilized Cells and Enzymes (IRL Press (1986)); Perbal (1984) A Practical Guide to Molecular Cloning; Miller and Calos eds. (1987) Gene Transfer Vectors for Mammalian Cells (Cold Spring Harbor Laboratory); Makrides ed. (2003) Gene Transfer and Expression in Mammalian Cells; Mayer and Walker eds. (1987) Immunochemical Methods in Cell and Molecular Biology (Academic Press, London); Herzenberg et al. eds (1996) Weir’s Handbook of Experimental Immunology; Manipulating the Mouse Embryo: A Laboratory Manual, 3rd edition (Cold Spring Harbor Laboratory Press (2002)); Sohail (ed.) (2004) Gene Silencing by RNA Interference: Technology and Application (CRC Press).

[0051] As used in the specification and claims, the singular form “a,” “an” and “the” include plural references unless the context clearly dictates otherwise. For example, the term “a cell” includes a plurality of cells, including mixtures thereof.

[0052] As used herein, the term “comprising” is intended to mean that the compounds, agents, compositions and methods include the recited elements, but not exclude others. “Consisting essentially of” when used to define compounds, agents, compositions and methods, shall mean excluding other elements of any essential significance to the combination. Thus, a composition consisting essentially of the elements as defined herein would not exclude trace contaminants, e.g., from the isolation and purification method and pharmaceutically acceptable carriers, preservatives, and the like. “Consisting of” shall mean excluding more than trace elements of other ingredients. Embodiments defined by each of these transition terms are within the scope of this technology. 11 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0053] All numerical designations, e.g., pH, temperature, time, concentration, and molecular weight, including ranges, are approximations which are varied (+) or (-) by increments of 1, 5, or 10%. It is to be understood, although not always explicitly stated that all numerical designations are preceded by the term “about.” It also is to be understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.

[0054] The term “about,” as used herein when referring to a measurable value such as an amount or concentration and the like, is meant to encompass variations of 20%, 10%, 5%, 1 %, 0.5%, or even 0.1 % of the specified amount.

[0055] As used herein, comparative terms as used herein, such as high, low, increase, decrease, reduce, or any grammatical variation thereof, can refer to certain variation from the reference. In some embodiments, such variation can refer to about 10%, or about 20%, or about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 80%, or about 90%, or about 1 fold, or about 2 folds, or about 3 folds, or about 4 folds, or about 5 folds, or about 6 folds, or about 7 folds, or about 8 folds, or about 9 folds, or about 10 folds, or about 20 folds, or about 30 folds, or about 40 folds, or about 50 folds, or about 60 folds, or about 70 folds, or about 80 folds, or about 90 folds, or about 100 folds or more higher than the reference. In some embodiments, such variation can refer to about 1%, or about 2%, or about 3%, or about 4%, or about 5%, or about 6%, or about 7%, or about 8%, or about 0%, or about 10%, or about 20%, or about 30%, or about 40%, or about 50%, or about 60%, or about 70%, or about 75%, or about 80%, or about 85%, or about 90%, or about 95%, or about 96%, or about 97%, or about 98%, or about 99% of the reference.

[0056] As will be understood by one skilled in the art, for any and all purposes, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Furthermore, as will be understood by one skilled in the art, a range includes each individual member.

[0057] “Optional” or “optionally” means that the subsequently described circumstance may or may not occur, so that the description includes instances where the circumstance occurs and instances where it does not.

[0058] As used herein, “and / or” refers to and encompasses any and all possible combinations of one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative (“or”). 12 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0059] “Substantially” or “essentially” means nearly totally or completely, for instance, 95% or greater of some given quantity. In some embodiments, “substantially” or “essentially” means 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.9%.

[0060] The terms or “acceptable,” “effective,” or “sufficient” when used to describe the selection of any components, ranges, dose forms, etc. disclosed herein intend that said component, range, dose form, etc. is suitable for the disclosed purpose.

[0061] A “composition” is intended to mean a combination of active agent and another compound or composition, inert (for example, a detectable agent or label) or active, such as an adjuvant, diluent, binder, stabilizer, buffers, salts, lipophilic solvents, preservative, adjuvant or the like and include pharmaceutically acceptable carriers.

[0062] Carriers also include pharmaceutical excipients and additives proteins, peptides, amino acids, lipids, and carbohydrates (e.g., sugars, including monosaccharides, di-, tri, tetra- oligosaccharides, and oligosaccharides; derivatized sugars such as alditols, aldonic acids, esterified sugars and the like; and polysaccharides or sugar polymers), which can be present singly or in combination, comprising alone or in combination 1-99.99% by weight or volume. Exemplary protein excipients include serum albumin such as human serum albumin (HSA), recombinant human albumin (rHA), gelatin, casein, and the like. Representative amino acid components, which can also function in a buffering capacity, include alanine, arginine, glycine, arginine, betaine, histidine, glutamic acid, aspartic acid, cysteine, lysine, leucine, isoleucine, valine, methionine, phenylalanine, aspartame, and the like. Carbohydrate excipients are also intended within the scope of this technology, examples of which include but are not limited to monosaccharides such as fructose, maltose, galactose, glucose, D- mannose, sorbose, and the like; disaccharides, such as lactose, sucrose, trehalose, cellobiose, and the like; polysaccharides, such as raffinose, melezitose, maltodextrins, dextrans, starches, and the like; and alditols, such as mannitol, xylitol, maltitol, lactitol, xylitol sorbitol (glucitol) and myoinositol.

[0063] A composition as disclosed herein can be a pharmaceutical composition. A “pharmaceutical composition” is intended to include the combination of an active agent with a carrier, inert or active, making the composition suitable for diagnostic or therapeutic use in vitro, in vivo or ex vivo.

[0064] “Pharmaceutically acceptable carriers” refers to any diluents, excipients, or carriers that may be used in the compositions disclosed herein. Pharmaceutically acceptable carriers 13 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 include ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. Suitable pharmaceutical carriers are described in Remington's Pharmaceutical Sciences, Mack Publishing Company, a standard reference text in this field. They may be selected with respect to the intended form of administration, that is, oral tablets, capsules, elixirs, syrups and the like, and consistent with conventional pharmaceutical practices.

[0065] The compositions used in accordance with the disclosure can be packaged in dosage unit form for ease of administration and uniformity of dosage. The term "unit dose" or "dosage" refers to physically discrete units suitable for use in a subject, each unit containing a predetermined quantity of the composition calculated to produce the desired responses in association with its administration, i.e., the appropriate route and regimen. The quantity to be administered, both according to number of treatments and unit dose, depends on the result and / or protection desired. Precise amounts of the composition also depend on the judgment of the practitioner and are peculiar to each individual. Factors affecting dose include physical and clinical state of the subject, route of administration, intended goal of treatment (alleviation of symptoms versus cure), and potency, stability, and toxicity of the particular composition. Upon formulation, solutions are administered in a manner compatible with the dosage formulation and in such amount as is therapeutically or prophylactically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described herein.

[0066] A combination as used herein intends that the individual active ingredients of the compositions are separately formulated for use in combination and can be separately packaged with or without specific dosages. The active ingredients of the combination can be administered concurrently or sequentially.

[0067] An “effective amount” is an amount sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications, or dosages. Such delivery is dependent on a number of variables including the time period for 14 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 which the individual dosage unit is to be used, the bioavailability of the therapeutic agent, the route of administration, etc. It is understood, however, that specific dose levels of the therapeutic agents disclosed herein for any particular subject depends upon a variety of factors including the activity of the specific agent employed, bioavailability of the agent, the route of administration, the age of the animal and its body weight, general health, sex, the diet of the animal, the time of administration, the rate of excretion, the drug combination, and the severity of the particular disorder being treated and form of administration. In general, one will desire to administer an amount of the agent that is effective to achieve a serum level commensurate with the concentrations found to be effective in vivo. These considerations, as well as effective formulations and administration procedures are well known in the art and are described in standard textbooks.

[0068] “Therapeutically effective amount” of an agent refers to an amount of the agent that is an amount sufficient to obtain a pharmacological response; or alternatively, is an amount of the agent that, when administered to a patient with a specified disorder or disease, is sufficient to have the intended effect, e.g., treatment, alleviation, amelioration, palliation or elimination of one or more manifestations of the specified disorder or disease in the patient. A therapeutic effect does not necessarily occur by administration of one dose and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations.

[0069] As used herein, the phrase “derived from” means isolated from, purified from, or engineered from, or any combination thereof.

[0070] As used herein, “treating” or “treatment” of a disease in a subject refers to (1) preventing the symptoms or disease from occurring in a subject that is predisposed or does not yet display symptoms of the disease; (2) inhibiting the disease or arresting its development; or (3) ameliorating or causing regression of the disease or the symptoms of the disease. As understood in the art, “treatment” is an approach for obtaining beneficial or desired results, including clinical results. For the purposes of the present technology, beneficial or desired results can include one or more, but are not limited to, alleviation or amelioration of one or more symptoms, diminishment of extent of a condition (including a disease), stabilized (i.e., not worsening) state of a condition (including disease), delay or slowing of condition (including disease), progression, amelioration or palliation of the condition (including disease), states and remission (whether partial or total), whether detectable or undetectable. When the disease is cancer, the following clinical end points are 15 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 non-limiting examples of treatment: reduction in tumor burden, slowing of tumor growth, longer overall survival, longer time to tumor progression, inhibition of metastasis or a reduction in metastasis of the tumor. In one aspect, treatment excludes prophylaxis.

[0071] As used herein, by "subject" or "individual" or "animal" or "patient" or "mammal," is meant any subject, particularly a mammalian subject, for whom diagnosis, prognosis, or therapy is desired. Mammalian subjects include, but are not limited to, humans, domestic animals, farm animals, zoo animals, sport animals, pet animals such as dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, cows; primates such as apes, monkeys, orangutans, and chimpanzees; canids such as dogs and wolves; felids such as cats, lions, and tigers; equids such as horses, donkeys, and zebras; bears, food animals such as cows, pigs, and sheep; ungulates such as deer and giraffes; rodents such as mice, rats, hamsters and guinea pigs; and so on. In certain embodiments, the mammal is a human subject. In other embodiments, a subject is a human patient. In a particular embodiment, a subject is a human patient in need of treatment.

[0072] In one embodiment, the term “disease” or “disorder” as used herein refers to any alteration in state of the body or of some of the organs, interrupting or disturbing the performance of the functions and / or causing symptoms such as discomfort, dysfunction, distress, or even death to the person afflicted or those in contact with a person.

[0073] The term “contacting” means direct or indirect binding or interaction between two or more. A particular example of direct interaction is binding. A particular example of an indirect interaction is where one entity acts upon an intermediary molecule, which in turn acts upon the second referenced entity. Contacting as used herein includes in solution, in solid phase, in vitro, ex vivo, in a cell and in vivo. Contacting in vivo can be referred to as administering, or administration.

[0074] “Administration” or “delivery” of an oncolytic virus or a composition containing same can be performed in one dose, continuously or intermittently throughout the course of treatment. Methods of determining the most effective means and dosage of administration are known to those of skill in the art and will vary with the composition used for therapy, the purpose of the therapy, the target cell being treated, and the subject being treated. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician or in the case of animals, by the treating veterinarian. Suitable dosage formulations and methods of administering the agents are known in the art. Route of 16 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 administration can also be determined and method of determining the most effective route of administration are known to those of skill in the art and will vary with the composition used for treatment, the purpose of the treatment, the health condition or disease stage of the subject being treated, and target cell or tissue. Non-limiting examples of route of administration include oral administration, intraperitoneal, infusion, nasal administration, inhalation, injection, and topical application. In some embodiments, the administration is administration to a tumor microenvironment. In some embodiments, administering or a grammatical variation thereof also refers to more than one doses with certain interval. In some embodiments, the interval is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 10 days, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 1 year or longer. In some embodiments, one dose is repeated for once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times or more.

[0075] The term administration shall include without limitation, administration by oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous, intravascular, intraperitoneal, intracerebroventricular (ICV), intrathecal, intracisternal injection or infusion, intracranial, ocular, intradermally, percutaneously, subcutaneous injection, or implant), intratumorally, by inhalation spray nasal, intratracheal, vaginal, rectal, sublingual, urethral (e.g., urethral suppository) or topical routes of administration (e.g., gel, ointment, cream, aerosol, etc.) and can be formulated, alone or together, in suitable dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants, excipients, and vehicles appropriate for each route of administration. The disclosure is not limited by the route of administration, the formulation or dosing schedule.

[0076] An agent of the present disclosure can be administered for therapy by any suitable route of administration. It will also be appreciated that the optimal route will vary with the condition and age of the recipient, and the disease being treated.

[0077] Administration or treatment in “combination” refers to administering two agents such that their pharmacological effects are manifest at the same time. Combination does not require administration at the same time or substantially the same time, although combination can include such administrations.

[0078] The phrase “first line” or “second line” or “third line” refers to the order of treatment received by a patient. First line therapy regimens are treatments given first, whereas second 17 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 or third line therapy are given after the first line therapy or after the second line therapy, respectively.

[0079] The terms "oligonucleotide" or "polynucleotide" or "portion," or "segment" thereof refer to a stretch of polynucleotide residues which is long enough to use in PCR or various hybridization procedures to identify or amplify identical or related parts of mRNA or DNA molecules. The polynucleotide compositions of this invention include RNA, cDNA, genomic DNA, synthetic forms, and mixed polymers, both sense and antisense strands, and may be chemically or biochemically modified or may contain non-natural or derivatized nucleotide bases, as will be readily appreciated by those skilled in the art. Such modifications include, for example, labels, methylation, substitution of one or more of the naturally occurring nucleotides with an analog, internucleotide modifications such as uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoamidates, carbamates, etc.), charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), pendent moieties (e.g., polypeptides), intercalators (e.g., acridine, psoralen, etc.), chelators, alkylators, and modified linkages (e.g., alpha anomeric nucleic acids, etc.). Also included are synthetic molecules that mimic polynucleotides in their ability to bind to a designated sequence via hydrogen bonding and other chemical interactions. Such molecules are known in the art and include, for example, those in which peptide linkages substitute for phosphate linkages in the backbone of the molecule.

[0080] As used herein, the term “purified” does not require absolute purity; rather, it is intended as a relative term. Thus, for example, a purified nucleic acid, peptide, protein, biological complexes, cell, virus or other active compound is one that is isolated in whole or in part from proteins or other contaminants. Generally, substantially purified peptides, proteins, biological complexes, cell, virus or other active compounds for use within the disclosure comprise more than 80% of all macromolecular species present in a preparation prior to admixture or formulation of the peptide, protein, biological complex, cell, virus or other active compound with a pharmaceutical carrier, excipient, buffer, absorption enhancing agent, stabilizer, preservative, adjuvant or other co-ingredient in a complete pharmaceutical formulation for therapeutic administration. More typically, the peptide, protein, biological complex, cell, virus or other active compound is purified to represent greater than 90%, often greater than 95% of all macromolecular species present in a purified preparation prior to admixture with other formulation ingredients. In other cases, the purified preparation may be 18 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 essentially homogeneous, wherein other macromolecular species are not detectable by conventional techniques.

[0081] In some embodiments, the term “engineered” or “recombinant” refers to having at least one modification not normally found in a naturally occurring protein, polypeptide, polynucleotide, strain, wild-type strain or the parental host strain of the referenced species. In some embodiments, the term “engineered” or “recombinant” refers to being synthetized by human intervention.

[0082] As used herein, the term “antibody” collectively refers to immunoglobulins or immunoglobulin-like molecules including by way of example and without limitation, IgA, IgD, IgE, IgG and IgM, combinations thereof, and similar molecules produced during an immune response in any vertebrate, for example, in mammals such as humans, goats, rabbits, camels and mice, as well as non-mammalian species, such as shark immunoglobulins. As used herein, “antibodies” (includes intact immunoglobulins) and “antigen binding fragments” specifically bind to a molecule of interest (or a group of highly similar molecules of interest) to the substantial exclusion of binding to other molecules (for example, antibodies and antibody fragments that have a binding constant for the molecule of interest that is at least 103 M-1 greater, at least 104 M-1 greater or at least 105 M-1 greater than a binding constant for other molecules in a biological sample). The term “antibody” also includes genetically engineered forms such as chimeric antibodies (for example, humanized murine antibodies), heteroconjugate antibodies (such as, bispecific antibodies). See also, Pierce Catalog and Handbook, 1994-1995 (Pierce Chemical Co., Rockford, Ill.); Kuby, J., Immunology, 3rd Ed., W.H. Freeman & Co., New York, 1997.

[0083] More particularly, antibody refers to a polypeptide ligand comprising at least a light chain immunoglobulin variable region or heavy chain immunoglobulin variable region which specifically recognizes and binds an epitope of an antigen. Antibodies are composed of a heavy and a light chain, each of which has a variable region, termed the variable heavy (VH) region and the variable light (VL) region. Together, the VH region and the VL region are responsible for binding the antigen recognized by the antibody. Typically, an immunoglobulin has heavy (H) chains and light (L) chains interconnected by disulfide bonds. There are two types of light chain, lambda (λ) and kappa (κ). There are five main heavy chain classes (or isotypes) which determine the functional activity of an antibody molecule: IgM, IgD, IgG, IgA and IgE. Each heavy and light chain contains a constant region and a variable region, (the regions are also known as “domains”). In combination, the heavy and the light 19 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 chain variable regions specifically bind the antigen. Light and heavy chain variable regions contain a “framework” region interrupted by three hypervariable regions, also called “complementarity-determining regions” or “CDRs”. The extent of the framework region and CDRs have been defined (see, Kabat et al., Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services, 1991, which is hereby incorporated by reference). The Kabat database is now maintained online. The sequences of the framework regions of different light or heavy chains are relatively conserved within a species. The framework region of an antibody, that is the combined framework regions of the constituent light and heavy chains, largely adopt a β-sheet conformation and the CDRs form loops which connect, and in some cases form part of, the β-sheet structure. Thus, framework regions act to form a scaffold that provides for positioning the CDRs in correct orientation by inter-chain, non-covalent interactions.

[0084] The CDRs are primarily responsible for binding to an epitope of an antigen. The CDRs of each chain are typically referred to as CDR1, CDR2, and CDR3, numbered sequentially starting from the N-terminus, and are also typically identified by the chain in which the particular CDR is located. Thus, a VH CDR3 is located in the variable domain of the heavy chain of the antibody in which it is found, whereas a VL CDR1 is the CDR1 from the variable domain of the light chain of the antibody in which it is found. An antibody that binds L1-CAM protein will have a specific VH region and the VL region sequence, and thus specific CDR sequences. Antibodies with different specificities (i.e., different combining sites for different antigens) have different CDRs. Although it is the CDRs that vary from antibody to antibody, only a limited number of amino acid positions within the CDRs are directly involved in antigen binding. These positions within the CDRs are called specificity determining residues (SDRs). “Immunoglobulin-related compositions” as used herein, refers to antibodies (including monoclonal antibodies, polyclonal antibodies, humanized antibodies, chimeric antibodies, recombinant antibodies, multispecific antibodies, bispecific antibodies, etc.,) as well as antibody fragments. An antibody or antigen binding fragment thereof specifically binds to an antigen.

[0085] As used herein, the term “antibody-related polypeptide” means antigen-binding antibody fragments, including single-chain antibodies, that can comprise the variable region(s) alone, or in combination, with all or part of the following polypeptide elements: hinge region, CH1, CH2, and CH3 domains of an antibody molecule. Also included in the technology are any combinations of variable region(s) and hinge region, CH1, CH2, and CH3 20 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 domains. Antibody-related molecules useful in the present methods, e.g., but are not limited to, Fab, Fab’ and F(ab’)2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide- linked Fvs (sdFv) and fragments comprising either a VL or VH domain. Examples include: (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab’)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al., Nature 341: 544-546, 1989), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR). As such “antibody fragments” or “antigen binding fragments” can comprise a portion of a full-length antibody, generally the antigen binding or variable region thereof. Examples of antibody fragments or antigen binding fragments include Fab, Fab', F(ab’)2, and Fv fragments; diabodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

[0086] "Bispecific antibody" or “BsAb”, as used herein, refers to an antibody that can bind simultaneously to two targets that have a distinct structure, e.g., two different target antigens, two different epitopes on the same target antigen, or a hapten and a target antigen or epitope on a target antigen. A variety of different bispecific antibody structures are known in the art. In some embodiments, each antigen binding moiety in a bispecific antibody includes VH and / or VL regions; in some such embodiments, the VH and / or VL regions are those found in a particular monoclonal antibody. In some embodiments, the bispecific antibody contains two antigen binding moieties, each including VH and / or VL regions from different monoclonal antibodies. In some embodiments, the bispecific antibody contains two antigen binding moieties, wherein one of the two antigen binding moieties includes an immunoglobulin molecule having VH and / or VL regions that contain CDRs from a first monoclonal antibody, and the other antigen binding moiety includes an antibody fragment (e.g., Fab, F(ab’), F(ab’)2, Fd, Fv, dAB, scFv, etc.) having VH and / or VL regions that contain CDRs from a second monoclonal antibody.

[0087] As used herein, the term “diabody” refers to a small antibody fragment with two antigen-binding sites, which fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create 21 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 two antigen binding sites. Diabodies are described more fully in, e.g., EP 404,097; WO 93 / 11161; and 30 Hollinger et al., Proc. Natl. Acad. Sci. USA, 90: 6444-6448 (1993).

[0088] As used herein, the terms “single-chain antibodies” or “single-chain Fv (scFv)” refer to an antibody fusion molecule of the two domains of the Fv fragment, VL and VH. Single- chain antibody molecules may comprise a polymer with a number of individual molecules, for example, dimer, trimer or other polymers. Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single- chain Fv (scFv)). Bird et al. (1988) Science 242:423-426 and Huston et al. (1988) Proc. Natl. Acad Sci. USA 85:5879-5883. Such single-chain antibodies can be prepared by recombinant techniques or enzymatic or chemical cleavage of intact antibodies.

[0089] Any of the above-noted antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for binding specificity and neutralization activity in the same manner as are intact antibodies.

[0090] As used herein, an “antigen” refers to a molecule to which an antibody (or antigen binding fragment thereof) can selectively bind. The target antigen may be a protein, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound. In some embodiments, the target antigen may be a polypeptide (e.g., a L1-CAM polypeptide). An antigen may also be administered to an animal to generate an immune response in the animal. Such antigens may also be called “immunogen”.

[0091] The term “antigen binding fragment” refers to a fragment of the whole immunoglobulin structure which possesses a part of a polypeptide responsible for binding to antigen. Examples of the antigen binding fragment useful in the present technology include scFv, (scFv)2, scFvFc, Fab, Fab’ and F(ab’)2, but are not limited thereto.

[0092] By “binding affinity” is meant the strength of the total noncovalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen, a hapten or antigenic peptide). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (Kd). Affinity can be measured by standard methods known in the art, including those described herein. A low-affinity complex contains an antibody that generally tends to dissociate readily from the antigen, whereas a 22 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 high-affinity complex contains an antibody that generally tends to remain bound to the antigen for a longer duration.

[0093] “Homology” or “identity” or “similarity” refers to sequence similarity between two peptides or between two nucleic acid molecules. Homology can be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When a position in the compared sequence is occupied by the same base or amino acid, then the molecules are homologous at that position. A degree of homology between sequences is a function of the number of matching or homologous positions shared by the sequences. A polynucleotide or polynucleotide region (or a polypeptide or polypeptide region) has a certain percentage (for example, at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99%) of “sequence identity” to another sequence means that, when aligned, that percentage of bases (or amino acids) are the same in comparing the two sequences. This alignment and the percent homology or sequence identity can be determined using software programs known in the art. In some embodiments, default parameters are used for alignment. One alignment program is BLAST, using default parameters. In particular, programs are BLASTN and BLASTP, using the following default parameters: Genetic code=standard; filter=none; strand=both; cutoff=60; expect=10; Matrix=BLOSUM62; Descriptions=50 sequences; sort by ═HIGH SCORE; Databases=non-redundant, GenBank+EMBL+DDBJ+PDB+GenBank CDS translations+SwissProtein+SPupdate+PIR. Details of these programs can be found at the National Center for Biotechnology Information. Biologically equivalent polynucleotides are those having the specified percent homology and encoding a polypeptide having the same or similar biological activity. Two sequences are deemed “unrelated” or “non-homologous” if they share less than 40% identity, or less than 25% identity, with each other. In this disclosure, wherever a percent identity is used in the context of an antibody sequence, the difference is in the regions outside the CDR regions (e.g., framework regions or structural regions) so the resulting antibody has the same affinity as the original antibody.

[0094] As used herein, the term “humanized” forms of non-human (e.g., murine) antibodies are chimeric antibodies which contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies are human immunoglobulins in which hypervariable region residues of the recipient are replaced by hypervariable region residues from a non-human species (donor antibody) such as mouse, rat, rabbit, camel or nonhuman primate having the desired specificity, affinity, and capacity. In some embodiments, Fv framework region (FR) residues of the human immunoglobulin are replaced 23 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues which are not found in the recipient antibody or in the donor antibody. These modifications are made to further refine antibody performance such as binding affinity. Generally, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains (e.g., Fab, Fab’, F(ab’)2, or Fv), in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus FR sequence although the FR regions may include one or more amino acid substitutions that improve binding affinity. The number of these amino acid substitutions in the FR are typically no more than 6 in the H chain, and in the L chain, no more than 3. The humanized antibody optionally may also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., Nature 321:522-525 (1986); Reichmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol.2:593-596 (1992). See e.g., Ahmed & Cheung, FEBS Letters 588(2):288- 297 (2014).

[0095] As used herein, the term “hypervariable region” refers to the amino acid residues of an antibody which are responsible for antigen-binding. The hypervariable region generally comprises amino acid residues from a “complementarity determining region” or “CDR” (e.g., around about residues 24-34 (L1), 50-56 (L2) and 89-97 (L3) in the VL, and around about 31-35B (H1), 50-65 (H2) and 95-102 (H3) in the VH (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)) and / or those residues from a “hypervariable loop” (e.g., residues 26- 32 (L1), 50-52 (L2) and 91-96 (L3) in the VL, and 26-32 (H1), 52A-55 (H2) and 96-101 (H3) in the VH (Chothia and Lesk J. Mol. Biol.196:901-917 (1987)).

[0096] As used herein, the terms “identical” or percent “identity”, when used in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region (e.g., nucleotide sequence encoding an antibody described herein or amino acid sequence of an antibody described herein)), when compared and aligned for maximum correspondence over a comparison window or designated region) as measured using a BLAST or BLAST 2.0 sequence comparison algorithms with default parameters described below, or by manual 24 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 alignment and visual inspection, e.g., NCBI web site). Such sequences are then said to be “substantially identical.” This term also refers to, or can be applied to, the complement of a test sequence. The term also includes sequences that have deletions and / or additions, as well as those that have substitutions. In some embodiments, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or 50-100 amino acids or nucleotides in length.

[0097] As used herein, the terms “individual,” “patient,” or “subject” can be an individual organism, a vertebrate, a mammal, or a human. In some embodiments, the individual, patient or subject is a human.

[0098] The term “monoclonal antibody” as used herein refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. For example, a monoclonal antibody can be an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope (aka. an “antigenic determinant”). Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. The modifier “monoclonal” indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. Monoclonal antibodies can be prepared using a wide variety of techniques known in the art including, e.g., but not limited to, hybridoma, recombinant, and phage display technologies. For example, the monoclonal antibodies to be used in accordance with the present methods may be made by the hybridoma method first described by Kohler et al., Nature 256:495 (1975), or may be made by recombinant DNA methods (See, e.g., U.S. Patent No.4,816,567). The “monoclonal antibodies” may also be isolated from phage antibody libraries using the techniques described in Clackson et al., Nature 352:624-628 (1991) and Marks et al., J. Mol. Biol.222:581-597 (1991), for example. 25 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0099] As used herein, the phrase “high density lipoprotein” or “HDL” refers to a lipid- protein complex which when isolated by ultracentrifugation is found in the density range of d = 1.063 to d = 1.21.

[0100] As used herein “ApoA-I Milano HDL” refers to a naturally occurring mutated variant of the apolipoprotein A1 protein found in human HDL, the lipoprotein particle that carries cholesterol from tissues to the liver and is associated with protection against cardiovascular disease. The ApoA-I protein on the Milano HDL has an arginine to cysteine substitution at position 173.

[0101] In some embodiments, an HDL molecule is a “reconstituted HDL” which refers to a complex of human Apolipoprotein A-I and 1-palmitoyl-2-oleoyl phosphatidylcholine prepared at a molar ratio of 1 to 100 by the cholate removal method. Reconstituted HDL is commercially available from RayBiotech, Cat. # MD-26-0020P, raybiotech.com / reconstituted-hdl-md-26-0020p, last accessed April 7, 2024; Fisher Scientific, Cat. # 15929708, fishersci.fi / shop / products / recombinant-human-hdl-protein- 3 / 15929708, last accessed April 7, 2024.

[0102] As used herein, “ApoA-I” (or “Apo A1”) refers to a gene encoding a lipid-binding protein (Apolipoprotein A-I) belonging to the apolipoprotein gene family. ApoA-I is the major protein component of high density lipoprotein (HDL) in plasma. (Genecards: APOA1, HGNC: 600, NCBI Gene: 335, Ensembl: ENSG00000118137, OMIM®: 107680, UniProtKB / Swiss-Prot: P02647).

[0103] As used herein, “ApoA-II” (or “Apo A2”) refers to a gene encoding the second most abundant protein of the high-density lipoprotein particles (Apolipoprotein A-II). (Genecards: APOA1, HGNC: 601, NCBI Gene: 336, Ensembl: ENSG00000158874, OMIM®: 107670, UniProtKB / Swiss-Prot: P02652).

[0104] As used herein, “ApoE” refers to a gene encoding a protein on the surface of HDL (“Apolipoprotein E” or “Apolipoprotein E3”). (Genecards: APOE, HGNC: 613, NCBI Gene: 348, Ensembl: ENSG00000130203, OMIM®: 107741, UniProtKB / Swiss-Prot: P02649).

[0105] As used herein, “ApoC-II” (or “Apo C2”) refers to a gene encoding a lipid-binding protein (Apolipoprotein C-II) belonging to the apolipoprotein gene family. (Genecards: APOC2, HGNC: 609, NCBI Gene: 344, Ensembl: ENSG00000234906, OMIM®: 608083, UniProtKB / Swiss-Prot: P02655). 26 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0106] As used herein, “HuR RNA Binding Protein” or “Human Antigen R” refers to gene is a member of the ELAVL family of RNA-binding proteins that contain several RNA recognition motifs. HuR RNA Binding Protein is encoded by the ELAVL1 gene. (Genecards: ELAVL1, HGNC: 3312, NCBI Gene: 1994, Ensembl: ENSG00000066044, OMIM®: 603466, UniProtKB / Swiss-Prot: Q15717).

[0107] As used herein, a “linker” refers to a chemical moiety that links two groups together. The linker can be cleavable or non-cleavable. Cleavable linkers can be hydrolyzable, enzymatically cleavable, pH sensitive, photolabile, or disulfide linkers, among others. Other linkers include homobifunctional (two identical functional groups) and heterobifunctional (two functional groups that are different from each other) linkers. A “linking group” is a functional group capable of forming a covalent linkage consisting of one or more bonds to a bioactive agent.

[0108] As used herein, a “hydrolyzable linker” refers to a chemical linkage or bond, such as a covalent bond, that undergoes hydrolysis under physiological conditions. The tendency of a bond to hydrolyze may depend not only on the general type of linkage connecting two central atoms between which the bond is severed, but also on the substituents attached to these central atoms. Non-limiting examples of hydrolytically susceptible linkages include esters of carboxylic acids, phosphate esters, acetals, ketals, acyloxyalkyl ether, imines, orthoesters, and some amide linkages.

[0109] As used herein, an “enzymatically cleavable linker” refers to a linkage that is subject to degradation by one or more enzymes. Some hydrolytically susceptible linkages may also be enzymatically degradable. For example, esterases may act on esters of carboxylic acid or phosphate esters, and proteases may act on peptide bonds and some amide linkages.

[0110] As used herein, a “pH sensitive linker” refers to a linkage that is stable at one pH and subject to degradation at another pH. For example, the pH sensitive linker can be stable at neutral or basic conditions, but labile at mildly acidic conditions.

[0111] As used herein, a “Photolabile linker” refers to a linkage, such as a covalent bond, that cleaves upon exposure to light. The photolabile linker includes an aromatic moiety in order to absorb the incoming light, which then triggers a rearrangement of the bonds in order to cleave the two groups linked by the photolabile linker.

[0112] As used herein, a “Self-immolative” or “double prodrug linker” refers to a linkage in which the main function of the linker is to release a functional agent only after selective 27 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 trigger activation (for example, a drop in pH or the presence of a tissue-specific enzyme) followed by spontaneous chemical breakdown to release the functional agent.

[0113] As used herein, “SpyCatcher-SpyTag system” refers to a protein ligation method based on a modified domain from a Streptococcus pyogenes surface protein (SpyCatcher), which recognizes a cognate 13-amino-acid peptide (SpyTag). Upon recognition, the two form a covalent isopeptide bond between the side chains of a lysine in SpyCatcher and an aspartate in SpyTag. (as described in Hatlem, Daniel, et al. "Catching a SPY: using the SpyCatcher- SpyTag and related systems for labeling and localizing bacterial proteins." International journal of molecular sciences 20.9 (2019): 2129, incorporated herein in its entirety).

[0114] As used herein, the terms “messenger RNA” or “mRNA” refer to any polynucleotide which encodes a polypeptide of interest and which is capable of being translated to produce the encoded polypeptide of interest in vitro, in vivo, in situ, or ex vivo. Traditionally, the basic components of an mRNA molecule include a coding region (“an open reading frame”), a 5’UTR, a 3’UTR, a 5’ cap, and a poly-A tail.

[0115] As used herein, the terms “modified messenger RNA” or “modified mRNA” refer to mRNA polynucleotides that include naturally occurring and / or non-naturally occurring modifications, for example, of a sugar, a nucleobase, or an internucleoside linkage (e.g., to a linking phosphate, to a phosphodiester linkage, or to the phosphodiester backbone). Non- natural modified nucleotides may be introduced during synthesis of post-synthesis of the polynucleotides to achieve desired functions or properties. The modifications may be present on an internucleoside linkage, purine or pyrimidine base, or sugar. The modification may be introduced with chemical synthesis or with a polymerase enzyme at the terminal of a chain or anywhere else in the chain. Any of the regions of a polynucleotide may be chemically modified.

[0116] As used herein, the term “open reading frame,” abbreviated as “ORF,” refers to a segment or region of an mRNA molecule that encodes a polypeptide. The ORF comprises a continuous stretch of non-overlapping, in-frame codons, beginning with the initiation codon and ending with a stop codon, and is translated by the ribosome.

[0117] As used herein, the term “sequence optimization” refers to a process or series of processes by which nucleobases in a reference nucleic acid sequence are replaced with alternative nucleobases, resulting in a nucleic acid sequence with improved properties, e.g., improved protein expression or decreased immunogenicity of the nucleic acid itself. 28 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0118] In general, the goal in sequence optimization is to produce a synonymous nucleotide sequence than encodes the same polypeptide sequence encoded by the reference nucleotide sequence. Thus, there are no amino acid substitutions (as a result of codon optimization) in the polypeptide encoded by the codon optimized nucleotide sequence with respect to the polypeptide encoded by the reference nucleotide sequence.

[0119] As used herein, the term “in vivo” refers to events that occur within an organism (e.g., animal, plant, or microbe or cell or tissue thereof). As used herein, the term “ex vivo” refers to events that occur outside of an organism (e.g., animal, plant, or microbe or cell or tissue thereof). Ex vivo events may take place in an environment minimally altered from a natural (e.g., in vivo) environment.

[0120] As used herein, “CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of X (X is a monoclonal antibody or an antibody sequence)” means that CDR-L1 is the same as CDR-L1 of X, CDR- L2 is the same as CDR-L2 of X, CDR-L3 is the same as CDR-L3 of X, CDR-H1 is the same as CDR-H1 of X, CDR-H2 is the same as CDR-H2 of X, and CDR-H3 is the same as CDR- H3 of X. Modes for Carrying Out the Disclosure

[0121] The present disclosure is based on the Applicant’s surprising finding that RNA can be efficiently delivered to a cell while tethered to the exterior of an HDL molecule. Applicant developed molecular hooks which can bind to a protein outside of an HDL molecule and RNA at the same time, effectively tethering the RNA to the exterior of the HDL molecule. Applicant surprisingly found that the disclosed molecular hooks protect RNA molecules from nucleases when complexed together. While conventional RNA delivery methods using lipid nanoparticles (LNPs) are unable to cross the blood-brain barrier, an RNA payload tethered to an HDL molecule using the disclosed molecular hooks can easily cross the blood-brain barrier and deliver RNA to previously inaccessible central nervous system cells and tissues. Molecular hooks

[0122] An aspect of the disclosure is directed to molecular hooks that are capable of tethering an RNA to an HDL molecule (“HDL”). The molecular hooks described herein can be used to transfer and deliver RNA into cells. The presently described molecular hooks are superior to the conventional methods of RNA delivery because (i) they can cross the blood-brain barrier 29 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 “hitchhiking” on HDL molecules; (ii) unlike lipid nanoparticle (LNP) delivery, they do not require ultra cold (e.g. -50oC) for storage or transport (which would save money and make RNA treatments easier to transport and store – improving patient accessibility to these therapies).

[0123] In some embodiments, the present molecular hooks are pre-loaded with an RNA of interest (mRNA, miRNA, siRNA, sRNA, lnRNA, anti-sense RNA, etc.), forming an RNA- molecular hook complex. When the RNA-molecular hook complex is administered to a subject, the complex will find and bind HDL molecules in the subject’s bloodstream. This is called “spontaneous self-assembly.” The HDL- RNA-molecular hook complex travels through the subject’s bloodstream and can traverse into, and deliver the RNA of interest into, many tissues including the central nervous system behind the blood-brain barrier.

[0124] In some embodiments, HDL-RNA-molecular hook complexes are prepared ex vivo, e.g., by mixing isolated HDL molecules with the molecular hook and the RNA of interest. In some embodiments, HDL-RNA-molecular hook complexes administered to a subject or contacted with a cell.

[0125] In some embodiments, the molecular hooks of the present disclosure are used in methods for delivering RNA to cells and organisms. In some embodiments, the molecular hooks comprise a plurality of antigen binding domains that bind to both RNA and HDL.

[0126] In some embodiments, the molecular hook comprises, or alternatively consists essentially of, or yet further consists of: a first antigen binding domain that specifically binds to an exterior facing protein of the HDL; and a second antigen binding domain that specifically binds to the ribonucleic acid (RNA).

[0127] In some embodiments, the first antigen binding domain is linked to the second antigen binding domain through a flexible linker. In some embodiments, the flexible linker comprises, or alternatively consists essentially of, or yet further consists of any one of SEQ ID NOs: 76-79. In some embodiments, the first antigen binding domain is an antibody, and the heavy chain of the first antigen binding domain is linked to the second antigen binding domain. In some embodiments, both the first antigen binding domain and the second antigen binding domain are each an antibody or a fragment thereof, and the heavy chain of the first antigen binding domain is linked to the heavy chain of the second antigen binding domain.

[0128] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises, or alternatively consists essentially of, or yet further consists of a HuR RNA 30 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 binding protein. In some embodiments, the HuR RNA binding protein comprises, or alternatively consists essentially of, or yet further consists of an amino acid sequence that is at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identical to SEQ ID NO: 1.

[0129] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

[0130] In some embodiments, the exterior facing protein is selected from any human apolipoprotein found on liposomes of any class, including but not limited to: Apolipoprotein B-100 (ApoB-100), Apolipoprotein B-48 (ApoB-48), Apolipoprotein C-I (ApoC-I), Apolipoprotein C-III (ApoC-III), Apolipoprotein D (ApoD), Apolipoprotein H (ApoH), Apolipoprotein L1 (ApoL1), Apolipoprotein M (ApoM), and Apolipoprotein(a) [Apo(a)]. Such apolipoproteins may be associated with high-density lipoproteins (HDL), low- density lipoproteins (LDL), very-low-density lipoproteins (VLDL), intermediate-density lipoproteins (IDL), or chylomicrons.

[0131] In some embodiments, the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C-II (Apo C-II).

[0132] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or a 31 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 monoclonal antibody produced from a hybridoma of accession No. NITE BP-02443. In some embodiments, the ApoA-I antibody is an antibody described in US20200018755A1, which is incorporated in its entirety. In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 .

[0133] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 67 and a heavy chain variable region has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 68. In this disclosure, wherever a percent identity is used in the context of an antibody sequence, the difference is in the regions outside the CDR regions.

[0134] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO: 68.

[0135] In some embodiments, the first antigen binding domain specifically binds ApoA-II and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of an ApoA-II antibody described in US20160245815A1, or the ApoA-II antibody EPR2913 from Abcam (Catalog No. ab92478, abcam.com / products / primary-antibodies / apolipoprotein-a- iiapoa-ii-antibody-epr2913-ab92478.html, last accessed April 8, 2024). In some embodiments, the first antigen binding domain that specifically binds to ApoA-II comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 32 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

[0136] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of an ApoE antibody described in US20190270794A1, or the ApoE3 antibody from Bio-techne R&D (Catalog No. MAB41445, bio-techne.com / p / antibodies / human-apolipoprotein-e3- apoe3-antibody-377109_mab41445, last accessed April 8, 2024).

[0137] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42.

[0138] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 41 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 42.

[0139] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further 33 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44.

[0140] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 43 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 44.

[0141] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region having at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 44.

[0142] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46.

[0143] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 45 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 46.

[0144] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region having at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 45 and a heavy chain variable region having 34 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 46.

[0145] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48.

[0146] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 47 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 48.

[0147] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50.

[0148] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 49 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 50.

[0149] In some embodiments, the first antigen binding domain that specifically binds to ApoE comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting 35 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 51 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 55 SEQ ID NO: 56 SEQ ID NO: 57 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 58 SEQ ID NO: 59 SEQ ID NO: 60 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 61 SEQ ID NO: 62 SEQ ID NO: 57 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 63 SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 66 SEQ ID NO: 62

[0150] In some embodiments, the first antigen binding domain specifically binds ApoC-II and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of 3E4 monoclonal ApoC-II antibody from Thermo Fisher® (Catalog No. H00000344-M01, thermofisher.com / antibody / product / APOC2-Antibody-clone-3E4-Monoclonal / H00000344- M01, last accessed April 8, 2024). Alternative sources of ApoC-II antibodies which can be used in the present molecular hooks and methods include: LS Bio (Catalog No. LS-B10603- 0.25, lsbio.com / antibodies / ihc-plus-apoc2-antibody-apolipoprotein-c-ii-antibody-elisa-ihc-ip- wb-western-ls-b10603 / 295268?trid=247, last accessed April 8, 2024), R&D Systems (Catalog No. AF4497-SP, rndsystems.com / products / human-apolipoprotein-c-ii-apoc2- antibody_af4497, last accessed April 8, 2024), OriGene® (Catalog No. TA357157, origene.com / catalog / antibodies / primary-antibodies / ta357157 / apolipoprotein-cii-apoc2-rabbit- polyclonal-antibody, last accessed April 8, 2024), and other antibodies listed in Genecards APOC2 page at genecards.org / cgi-bin / carddisp.pl?gene=APOC2, last accessed April 8, 2024).

[0151] In some embodiments, the first antigen binding domain of the molecular hook comprises an anti-APOA-I antibody linked and the second antigen binding domain of the molecular hook comprises an RNA binding molecule, wherein the heavy chain of the anti- APOA-I antibody is linked to the RNA binding molecule by a flexible linker. In some embodiments, the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or 36 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 more, or any value therebetween) identity to SEQ ID NO: 68, a light chain having at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain and the HuR RNA binding protein are linked together by any one of flexible linkers as shown by SEQ ID NOs: 76-79. In some embodiments, the flexible linker comprises, consists of, or consists essentially of, or yet further consists of a sequence as shown by SEQ ID NO: 78. In some embodiments, the molecular hook comprises, or alternatively consists essentially of, or yet further consists of SEQ ID NO: 75 (anti-APOA-I heavy chain linked to HuR RNA binding protein) and SEQ ID NO: 67 (anti-APOA-I light chain).

[0152] In some embodiments, the first antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody. In some embodiments, the second antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody.

[0153] In certain embodiments, the first and / or second antigen-binding domain may comprise structures including but not limited to: a single-chain variable fragment (scFv), a tandem scFv dimer [(scFv)₂], an scFv-Fc fusion, a Fab, a Fab′, an F(ab′)₂ fragment, a bispecific antibody, a multispecific antibody, a diabody, or Fc-fusion proteins, as well as isotypes, engineered variants, or functional fragments thereof derived from such formats.

[0154] In some embodiments, the molecular hook comprises, or alternatively consists essentially of, or yet further consists of a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker. In some embodiments, the linker between the first antigen binding domain and the second antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of a linker from Spycatcher-Spytag system.

[0155] Additionally, in certain embodiments, the first antibody and the second antibody, and / or the HuR protein bound to the first antibody, are not only designed to bind one another post-expression, but may also be co-expressed as a single fusion construct, such that they are genetically and translationally linked during recombinant production. In certain embodiments, the molecular hook comprises a linker between a first antigen-binding domain 37 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 and a second antigen-binding domain, or alternatively, essentially consists of or consists entirely of such a linker. Optionally, the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH-sensitive linker, a photolabile linker, or a self-immolative linker. In some embodiments, the linker between the first and second antigen-binding domains comprises, or essentially consists of, a SpyCatcher–SpyTag system. Additionally, in certain embodiments, the first antibody and the second antibody, and / or the HuR protein bound to the first antibody, are not only designed to bind one another post-expression, but may also be co-expressed as a single fusion construct, such that they are genetically and translationally linked during recombinant production.

[0156] In some embodiments, the molecular hook further comprises a detectable label.

[0157] In another aspect, this disclosure provides polynucleotides encoding the molecular hooks as described herein. In some embodiments, the polynucleotides are DNA, RNA or a DNA-RNA hybrid. In some embodiments, the polynucleotides detectably labeled. Another aspect of the disclosure is directed to vectors and host cells comprising the polynucleotides encoding the molecular hooks. In some embodiments, the host cells are prokaryotic or eukaryotic. Methods for delivering Ribonucleic acid (RNA) into cells and Methods of Treatment.

[0158] An aspect of the disclosure is directed to an in vivo method for delivering ribonucleic acid (RNA) to cells of a subject in need thereof comprising, or alternatively consisting essentially of, or yet further consisting of administering to the subject an RNA-HDL complex wherein the RNA is tethered to the exterior of a high-density lipoprotein (HDL).

[0159] Another aspect of the disclosure is directed to an in vivo method for delivering ribonucleic acid (RNA) to cells of a subject in need thereof comprising, or alternatively consisting essentially of, or yet further consisting of administering to the subject an RNA- molecular hook complex wherein the molecular hook is as described herein and is capable of tethering to the exterior of any HDL molecule.

[0160] Another aspect of the disclosure is directed to a method for treating a subject in need thereof comprising, or alternatively consisting essentially of, or yet further consisting of administering to the subject an RNA-molecular hook complex wherein the molecular hook is as described herein and is capable of tethering to the exterior of any HDL molecule. 38 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0161] The molecular hooks and / or RNA-HDL complexes as described herein are useful in the preparation of medicaments to deliver RNA to cells and treat various diseases as described herein.

[0162] In some embodiments, the subject suffers from a single gene disorder (i.e., the disease is a disease caused by a mutation in one gene) and the RNA delivered on HDL using molecular hooks corrects or compensates for the mutation in the single gene. In some embodiments, the disease is selected from spinal muscular atrophy (SMA), Cystic fibrosis, Marfan syndrome, Huntington’s Disease, alpha thalassemia, beta thalassemia, sickle cell anemia, Fragile X syndrome, or hemochromatosis.. In some embodiments, the subject suffers from cancer, malaria, tuberculosis, Hepatitis A, Hepatitis B, Hepatitis C, HIV / AIDS, rabies, influenza, or COVID-19.

[0163] Another aspect of the disclosure is directed to a method for delivering ribonucleic acid (RNA) to a cell comprising, or alternatively consisting essentially of, or yet further consisting of contacting the cells with an RNA-HDL complex wherein the RNA is tethered to the exterior of an HDL. In some embodiments, the contacting is performed in vitro or in vivo. In some embodiments, the RNA is tethered to the exterior of an HDL through a molecular hook, wherein the molecular hook is as described herein. In some embodiments, the binding of RNA to the molecular hook prevents the degradation of the RNA during transport and delivery to a target cell or tissue. In some embodiments, the binding of RNA to the molecular hook protects the cargo RNA from degradation during transport and delivery to a target cell or tissue.

[0164] In some embodiments, the HDL molecule is the native HDL of a subject. In some embodiments, the HDL molecule is selected from a native HDL, an ApoA-I Milano HDL, or a reconstituted HDL.

[0165] In some embodiments, the cells of the subject are behind the blood brain barrier of the subject. In some embodiments, the cells of the subject are in the brain or the spine of the subject.

[0166] In some embodiments, the RNA comprises, or alternatively consists essentially of, or yet further consists of miRNA, siRNA, sRNA, lnRNA, or anti-sense RNA.

[0167] In some embodiments, the RNA comprises, or alternatively consists essentially of, or yet further consists of microRNA (miRNA), small interfering RNA (siRNA), small RNA (sRNA), long non-coding RNA (lncRNA), antisense RNA, messenger RNA (mRNA), guide 39 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 RNA (gRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), circular RNA (circRNA), ribosomal RNA (rRNA), transfer RNA (tRNA), double-stranded RNA (dsRNA), or any other naturally occurring, chemically modified, synthetically engineered, or as-yet- undiscovered RNA species that are capable of performing a therapeutic, diagnostic, regulatory, structural, or gene-modulatory function.

[0168] In some embodiments, the RNA comprises, or alternatively consists essentially of, or yet further consists of mRNA, and optionally wherein the mRNA encodes a therapeutic protein; and optionally wherein the mRNA encodes a vaccine. In some embodiments, the mRNA is not a modified mRNA (i.e., does not contain any modifications). In some embodiments, the mRNA is sequence optimized. In some embodiments, the mRNA is sequence optimized for improved expression in a target tissue. In some embodiments, the mRNA is sequence optimized for improved stability.

[0169] In some embodiments, the molecular hook comprises, or alternatively consists essentially of, or yet further consists of: a first antigen binding domain that specifically binds to an exterior facing protein of the HDL; and a second antigen binding domain that specifically binds to the ribonucleic acid (RNA).

[0170] In some embodiments, the first antigen binding domain is linked to the second antigen binding domain through a flexible linker. In some embodiments, the flexible linker comprises, or alternatively consists essentially of, or yet further consists of any one of SEQ ID NOs: 76-79. In some embodiments, the first antigen binding domain is an antibody, and the heavy chain of the first antigen binding domain is linked to the second antigen binding domain. In some embodiments, both the first antigen binding domain and the second antigen binding domain are each an antibody or a fragment thereof, and the heavy chain of the first antigen binding domain is linked to the heavy chain of the second antigen binding domain.

[0171] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises, or alternatively consists essentially of, or yet further consists of a HuR RNA binding protein. In some embodiments, the HuR RNA binding protein comprises, or alternatively consists essentially of, or yet further consists of an amino acid sequence that is at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identical to SEQ ID NO: 1.

[0172] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, 40 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

[0173] In some embodiments, the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C-II (Apo C-II).

[0174] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02443. In some embodiments, the ApoA-I antibody is an antibody described in US20200018755A1, which is incorporated in its entirety. In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 .

[0175] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 67 and a heavy chain variable region has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 68. In this disclosure, wherever a percent identity is used in the context of an antibody sequence, the difference is in the regions outside the CDR regions. 41 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0176] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO: 68.

[0177] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises, or alternatively consists essentially of, or yet further consists of a light chain sequence comprising, or alternatively consisting essentially of, or yet further consisting of SEQ ID NO: 67. In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises, or alternatively consists essentially of, or yet further consists of a heavy chain sequence comprising, or alternatively consisting essentially of, or yet further consisting of SEQ ID NO: 68.

[0178] In some embodiments, the first antigen binding domain that specifically binds to ApoA-II comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 .

[0179] In some embodiments, the first antigen binding domain specifically binds ApoA-II and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of an ApoA-II antibody described in US20160245815A1, or the ApoA-II antibody EPR2913 from Abcam (Catalog No. ab92478, abcam.com / products / primary-antibodies / apolipoprotein-a- iiapoa-ii-antibody-epr2913-ab92478.html, last accessed April 8, 2024). In some embodiments, the first antigen binding domain that specifically binds to ApoA-II comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 42 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

[0180] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of an ApoE antibody described in US20190270794A1, or the ApoE3 antibody from Bio-techne R&D (Catalog No. MAB41445, bio-techne.com / p / antibodies / human-apolipoprotein-e3- apoe3-antibody-377109_mab41445, last accessed April 8, 2024).

[0181] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42.

[0182] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 41 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 42.

[0183] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further 43 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44.

[0184] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 43 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 44.

[0185] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46.

[0186] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 45 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 46.

[0187] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48.

[0188] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 47 and a heavy chain variable region that 44 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 48.

[0189] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50.

[0190] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 49 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 50.

[0191] In some embodiments, the first antigen binding domain that specifically binds to ApoE comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 51 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 55 SEQ ID NO: 56 SEQ ID NO: 57 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 58 SEQ ID NO: 59 SEQ ID NO: 60 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 61 SEQ ID NO: 62 SEQ ID NO: 57 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 63 SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 66 SEQ ID NO: 62

[0192] In some embodiments, the first antigen binding domain specifically binds ApoC-II and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as 45 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of 3E4 monoclonal ApoC-II antibody from Thermo Fisher (Catalog No. H00000344-M01, thermofisher.com / antibody / product / APOC2-Antibody-clone-3E4-Monoclonal / H00000344- M01, last accessed April 8, 2024). Alternative sources of ApoC-II antibodies which can be used in the present molecular hooks and methods include: LS Bio (Catalog No. LS-B10603- 0.25, lsbio.com / antibodies / ihc-plus-apoc2-antibody-apolipoprotein-c-ii-antibody-elisa-ihc-ip- wb-western-ls-b10603 / 295268?trid=247, last accessed April 8, 2024), R&D Systems (Catalog No. AF4497-SP, rndsystems.com / products / human-apolipoprotein-c-ii-apoc2- antibody_af4497, last accessed April 8, 2024), OriGene (Catalog No. TA357157, origene.com / catalog / antibodies / primary-antibodies / ta357157 / apolipoprotein-cii-apoc2-rabbit- polyclonal-antibody, last accessed April 8, 2024), and other antibodies listed in Genecards APOC2 page at genecards.org / cgi-bin / carddisp.pl?gene=APOC2, last accessed April 8, 2024).

[0193] In some embodiments, the first antigen binding domain of the molecular hook comprises an anti-APOA-I antibody linked and the second antigen binding domain of the molecular hook comprises an RNA binding molecule, wherein the heavy chain of the anti- APOA-I antibody is linked to the RNA binding molecule by a flexible linker. In some embodiments, the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain and the HuR RNA binding protein are linked together by any one of flexible linkers as shown by SEQ ID NOs: 76-79. In some embodiments, the flexible linker comprises, consists of, or consists essentially of, or yet further consists of a sequence as shown by SEQ ID NO: 78. In some embodiments, the molecular hook comprises, or alternatively consists essentially of, or yet further consists of SEQ ID NO: 75 (anti-APOA-I heavy chain linked to HuR RNA binding protein) and SEQ ID NO: 67 (anti-APOA-I light chain).

[0194] In some embodiments, the first antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody. In some embodiments, the second antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody. 46 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0195] In certain embodiments, the first and / or second antigen-binding domain may comprise structures including but not limited to: a single-chain variable fragment (scFv), a tandem scFv dimer [(scFv)₂], an scFv-Fc fusion, a Fab, a Fab′, an F(ab′)₂ fragment, a bispecific antibody, a multispecific antibody, a diabody, or Fc-fusion proteins, as well as isotypes, engineered variants, or functional fragments thereof derived from such formats.

[0196] In some embodiments, the molecular hook comprises, or alternatively consists essentially of, or yet further consists of a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker. In some embodiments, the linker between the first antigen binding domain and the second antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of a linker from Spycatcher-Spytag system.

[0197] In some embodiments, the molecular hook further comprises a detectable label. Uses of RNA-HDL complexes and Molecular Hooks

[0198] Another aspect of the disclosure is directed to use of an RNA-HDL complex for the manufacture of a medicament for delivering ribonucleic acid (RNA) to cells, wherein the RNA-HDL complex comprises, or alternatively consists essentially of, or yet further consists of an RNA tethered to the exterior of a high-density lipoprotein (HDL).

[0199] Another aspect of the disclosure is directed to use of an RNA-molecular hook complex for the manufacture of a medicament for delivering ribonucleic acid (RNA) to cells, wherein the molecular hook is as described herein and is capable of tethering to the exterior of any HDL molecule.

[0200] The molecular hooks and / or RNA-HDL complexes as described herein are useful in the preparation of medicaments to deliver RNA to cells and treat various diseases as described herein.

[0201] In some embodiments, the medicament is for treating a single gene disorder (i.e., the disease is a disease caused by a mutation in one gene) and the RNA tethered to the exterior of a high-density lipoprotein (HDL) using a molecular hook corrects or compensates for the mutation in the single gene. In some embodiments, the disorder is selected from spinal muscular atrophy (SMA), Cystic fibrosis, Marfan syndrome, Huntington’s Disease, alpha thalassemia, beta thalassemia, sickle cell anemia, Fragile X syndrome, or hemochromatosis.. 47 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 In some embodiments, the disorder is selected from cancer, malaria, tuberculosis, Hepatitis A, Hepatitis B, Hepatitis C, HIV / AIDS, rabies, influenza, or COVID-19.

[0202] In some embodiments, the binding of RNA to the molecular hook prevents the degradation of the RNA during transport and delivery to a target cell or tissue. In some embodiments, the binding of RNA to the molecular hook protects the cargo RNA from degradation during transport and delivery to a target cell or tissue.

[0203] In some embodiments, the HDL molecule is the native HDL of a subject. In some embodiments, the HDL molecule is selected from a native HDL, an ApoA-I Milano HDL, or a reconstituted HDL.

[0204] In some embodiments, the RNA comprises, or alternatively consists essentially of, or yet further consists of miRNA, siRNA, sRNA, lnRNA, or anti-sense RNA.

[0205] In some embodiments, the RNA comprises, or alternatively consists essentially of, or yet further consists of microRNA (miRNA), small interfering RNA (siRNA), small RNA (sRNA), long non-coding RNA (lncRNA), antisense RNA, messenger RNA (mRNA), guide RNA (gRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), circular RNA (circRNA), ribosomal RNA (rRNA), transfer RNA (tRNA), double-stranded RNA (dsRNA), or any other naturally occurring, chemically modified, synthetically engineered, or as-yet- undiscovered RNA species that are capable of performing a therapeutic, diagnostic, regulatory, structural, or gene-modulatory function.

[0206] In some embodiments, the RNA comprises, or alternatively consists essentially of, or yet further consists of mRNA, and optionally wherein the mRNA encodes a therapeutic protein; and optionally wherein the mRNA encodes a vaccine. In some embodiments, the mRNA is not a modified mRNA (i.e., does not contain any modifications). In some embodiments, the mRNA is sequence optimized. In some embodiments, the mRNA is sequence optimized for improved expression in a target tissue. In some embodiments, the mRNA is sequence optimized for improved stability.

[0207] In some embodiments, the molecular hook comprises, or alternatively consists essentially of, or yet further consists of: a first antigen binding domain that specifically binds to an exterior facing protein of the HDL; and a second antigen binding domain that specifically binds to the ribonucleic acid (RNA).

[0208] In some embodiments, the first antigen binding domain is linked to the second antigen binding domain through a flexible linker. In some embodiments, the flexible linker 48 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 comprises, or alternatively consists essentially of, or yet further consists of any one of SEQ ID NOs: 76-79. In some embodiments, the first antigen binding domain is an antibody, and the heavy chain of the first antigen binding domain is linked to the second antigen binding domain. In some embodiments, both the first antigen binding domain and the second antigen binding domain are each an antibody or a fragment thereof, and the heavy chain of the first antigen binding domain is linked to the heavy chain of the second antigen binding domain.

[0209] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises, or alternatively consists essentially of, or yet further consists of a HuR RNA binding protein. In some embodiments, the HuR RNA binding protein comprises, or alternatively consists essentially of, or yet further consists of an amino acid sequence that is at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identical to SEQ ID NO: 1.

[0210] In some embodiments, the second antigen binding domain that specifically binds to RNA comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

[0211] In some embodiments, the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C-II (Apo C-II).

[0212] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02443. In some 49 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 embodiments, the ApoA-I antibody is an antibody described in US20200018755A1, which is incorporated in its entirety. In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 .

[0213] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO: 68.

[0214] In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises, or alternatively consists essentially of, or yet further consists of a light chain sequence comprising, or alternatively consisting essentially of, or yet further consisting of SEQ ID NO: 67. In some embodiments, the first antigen binding domain specifically binds ApoA-I and comprises, or alternatively consists essentially of, or yet further consists of a heavy chain sequence comprising, or alternatively consisting essentially of, or yet further consisting of SEQ ID NO: 68.

[0215] In some embodiments, the first antigen binding domain that specifically binds to ApoA-II comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 .

[0216] In some embodiments, the first antigen binding domain specifically binds ApoA-II and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of an ApoA-II antibody described in US20160245815A1, or the ApoA-II antibody EPR2913 from 50 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 Abcam (Catalog No. ab92478, abcam.com / products / primary-antibodies / apolipoprotein-a- iiapoa-ii-antibody-epr2913-ab92478.html, last accessed April 8, 2024). In some embodiments, the first antigen binding domain that specifically binds to ApoA-II comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

[0217] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of an ApoE antibody described in US20190270794A1, or the ApoE3 antibody from Bio-techne R&D (Catalog No. MAB41445, bio-techne.com / p / antibodies / human-apolipoprotein-e3- apoe3-antibody-377109_mab41445, last accessed April 8, 2024).

[0218] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42.

[0219] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 41 and a heavy chain variable region that 51 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 42.

[0220] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44.

[0221] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 43 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 44.

[0222] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46.

[0223] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 45 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 46.

[0224] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47. In some embodiments, the first antigen binding domain 52 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48.

[0225] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 47 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 48.

[0226] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR- L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49. In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50.

[0227] In some embodiments, the first antigen binding domain specifically binds ApoE and comprises, or alternatively consists essentially of, or yet further consists of a light chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 49 and a heavy chain variable region that has at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 50.

[0228] In some embodiments, the first antigen binding domain that specifically binds to ApoE comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 51 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 55 SEQ ID NO: 56 SEQ ID NO: 57 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 58 SEQ ID NO: 59 SEQ ID NO: 60 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 61 SEQ ID NO: 62 53 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 SEQ ID NO: 57 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 63 SEQ ID NO: 64 SEQ ID NO: 65 SEQ ID NO: 52 SEQ ID NO: 53 SEQ ID NO: 54 SEQ ID NO: 66 SEQ ID NO: 62

[0229] In some embodiments, the first antigen binding domain specifically binds ApoC-II and comprises, or alternatively consists essentially of, or yet further consists of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising, or alternatively consisting essentially of, or yet further consisting of amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of 3E4 monoclonal ApoC-II antibody from Thermo Fisher (Catalog No. H00000344-M01, thermofisher.com / antibody / product / APOC2-Antibody-clone-3E4-Monoclonal / H00000344- M01, last accessed April 8, 2024). Alternative sources of ApoC-II antibodies which can be used in the present molecular hooks and methods include: LS Bio (Catalog No. LS-B10603- 0.25, lsbio.com / antibodies / ihc-plus-apoc2-antibody-apolipoprotein-c-ii-antibody-elisa-ihc-ip- wb-western-ls-b10603 / 295268?trid=247, last accessed April 8, 2024), R&D Systems (Catalog No. AF4497-SP, rndsystems.com / products / human-apolipoprotein-c-ii-apoc2- antibody_af4497, last accessed April 8, 2024), OriGene (Catalog No. TA357157, origene.com / catalog / antibodies / primary-antibodies / ta357157 / apolipoprotein-cii-apoc2-rabbit- polyclonal-antibody, last accessed April 8, 2024), and other antibodies listed in Genecards APOC2 page at genecards.org / cgi-bin / carddisp.pl?gene=APOC2, last accessed April 8, 2024).

[0230] In some embodiments, the first antigen binding domain of the molecular hook comprises an anti-APOA-I antibody linked and the second antigen binding domain of the molecular hook comprises an RNA binding molecule, wherein the heavy chain of the anti- APOA-I antibody is linked to the RNA binding molecule by a flexible linker. In some embodiments, the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 68, a light chain having at least 90% (e.g., 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% or more, or any value therebetween) identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain and the HuR RNA binding protein are linked together by any one of flexible linkers as shown by SEQ ID NOs: 76-79. In some embodiments, the flexible linker comprises, consists of, or consists essentially of, or yet further consists of a sequence as shown by SEQ ID NO: 78. In some embodiments, the 54 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 molecular hook comprises, or alternatively consists essentially of, or yet further consists of SEQ ID NO: 75 (anti-APOA-I heavy chain linked to HuR RNA binding protein) and SEQ ID NO: 67 (anti-APOA-I light chain).

[0231] In some embodiments, the first antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody. In some embodiments, the second antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody.

[0232] In certain embodiments, the first and / or second antigen-binding domain may comprise structures including but not limited to: a single-chain variable fragment (scFv), a tandem scFv dimer [(scFv)₂], an scFv-Fc fusion, a Fab, a Fab′, an F(ab′)₂ fragment, a bispecific antibody, a multispecific antibody, a diabody, or Fc-fusion proteins, as well as isotypes, engineered variants, or functional fragments thereof derived from such formats.

[0233] In some embodiments, the molecular hook comprises, or alternatively consists essentially of, or yet further consists of a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker. In some embodiments, the linker between the first antigen binding domain and the second antigen binding domain comprises, or alternatively consists essentially of, or yet further consists of a linker from Spycatcher-Spytag system.

[0234] Additionally, in certain embodiments, the first antibody and the second antibody, and / or the HuR protein bound to the first antibody, are not only designed to bind one another post-expression, but may also be co-expressed as a single fusion construct, such that they are genetically and translationally linked during recombinant production. In certain embodiments, the molecular hook comprises a linker between a first antigen-binding domain and a second antigen-binding domain, or alternatively, essentially consists of or consists entirely of such a linker. Optionally, the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH-sensitive linker, a photolabile linker, or a self-immolative linker. In some embodiments, the linker between the first and second antigen-binding domains comprises, or essentially consists of, a SpyCatcher–SpyTag system. Additionally, in certain embodiments, the first antibody and the second antibody, and / or the HuR protein bound to the first antibody, are not only designed to bind one another post-expression, but may also be 55 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 co-expressed as a single fusion construct, such that they are genetically and translationally linked during recombinant production.

[0235] In some embodiments, the molecular hook further comprises a detectable label.

[0236] The following examples are included to demonstrate some embodiments of the disclosure. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. EXAMPLES Example 1: Delivering RNA into cells using molecular hooks

[0237] Materials and Methods: Human serum containing naturally occurring ApoA1 and human nanolipid (HDL) particles were added at a concentration of 10%. HepG2 cell line was used.

[0238] HepG2 cells, as mentioned earlier, were cultured in 75 cm2cell culture flasks containing 15 ml Eagle's Minimal Essential Medium (MEM) supplemented with 10% human serum Type AB (Capricon Scientific Cat-No: HUM-3B), penicillin / streptomycin (Capricon Scientific Cat-No: PS-B).

[0239] To validate the concept, RNA encoding green fluorescent protein (GFP) was used as the RNA to be transported into the liver cells.

[0240] Preparing RNA capable of expressing GFP: A plasmid obtained from Addgene (pCAG-eCas9-GFP-U6-gRNA Addgene plasmid #79145) was transferred to suitable competent bacterial cells (DH5alpha) using the heat shock transformation method. Fifty microliters of competent cells were transferred sterilely into 1.5 ml Eppendorf tubes prechilled on ice. Two microliters of the prepared plasmid were added sterilely onto the bacterial cells for transformation. The tubes containing the mixture were incubated on ice for 5 minutes and then placed in a preheated 37°C heat block or water bath for 30 seconds. After this incubation period, the tubes were quickly placed back into ice for 2-3 minutes. The suspension was then adapted with LB medium.

[0241] Preparation of bacterial cultures in LB medium: Mix 10g Tryptone (Sigma-Aldrich, USA®), 5g yeast extract, and 10g NaCl (Sigma-Aldrich, USA) in a conical flask, and add 950 ml autoclaved distilled water (pH 7.0). Autoclave the mixture at 15 psi for 20 minutes. 56 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 After autoclaving and cooling, add antibiotics (ampicillin). The transformed bacteria adapted in the previous step were subjected to mini culture in an antibiotic medium. The bacteria cultured overnight at 1200 rpm and 37°C were centrifuged at the end of the culture.

[0242] The isolation of plasmid DNA from bacterial pellets was performed using the NucleoSpin Plasmid Miniprep Kit (MACHEREY-NAGEL GmbH & Co. KG). The concentration of plasmids and the A260 / A280 ratios were measured spectrophotometrically.

[0243] Preparation of functional molecular hooks. A fusion protein (Anti-Apo-A1-RNA Engagers (BAPs) was generated by fusing the HuR ribonucleoprotein—capable of binding, stabilizing, and transporting mRNAs such as GFP-mRNA—to the CH3 domain of the Fc fragment of an anti-ApoA-I antibody, which can associate with HDL, a natural lipid nanoparticle. The construct was produced using recombinant protein expression systems, such as [host system, e.g., E. coli], with standard transfection and / or expression protocols.

[0244] Measuring the transfection efficiency of molecular hooks. HepG2 hepatoma cells were transfected with the construct carrying Green Fluorescent Protein (GFP) mRNA using Bispecific Anti-Apo-A1-RNA Engagers (BAPs). The cells were cultured in 75 cm2cell culture flasks containing 15 ml Eagle's Minimal Essential Medium (MEM) supplemented with 10% human serum Type AB and penicillin / streptomycin in an atmosphere of 95% air and 5% CO2 at 37°C. Following transfection, the efficiency of GFP expression from the construct was planned to be measured by flow cytometry at 24 and 48 hours. Successfully transfected cells were harvested in a clean cell culture medium in a FACS tube and diluted with 1xHBSS solution. Flow cytometry was performed using the FITC channel within the wavelength range of 495 nm to 519 nm.

[0245] To determine whether HDL, which is human natural lipid nanoparticles (LNPs) and added to the medium, has a cytotoxic effect in cell culture medium, equal numbers of HepG2 cells were seeded in 6-well plates. After HepG2 cells were grown healthily in culture medium, they were treated with the construct. No cytotoxic effect was detected in the cells after treatment with the molecular hook construct. HepG2 cells continued to survive after application of the construct.

[0246] Results: Flow cytometry showed an average RNA delivery efficiency of about 10% at 96 hours after treatment, as judged by GFP expression in HepG2 cells (measured by flow cytometry), which is enough for RNA-based treatments and comparable to other in vivo RNA delivery methods such as human natural lipid nanoparticles (HDL) (FIG.2). 57 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 Example 2: Testing Delivery of Molecular hooks into Cells

[0247] The aim of the study was to assess the fluorescence intensity deriving from internalization of dye-conjugated ApoA-I antibody that is used in molecular hooks of the present disclosure. Increase of cell fluorescence indicates internalization, as the intracellular acidic conditions release the dye from the antibody, causing the rise in fluorescence. Increase of background fluorescence indicates acidification of the cell culture medium. PANC1 pancreatic cancer cell line was used in the tests comparing internalization of conjugated ApoA-I antibody and anti-HEL IgG as isotype control. PANC-1 cells are human pancreatic ductal adenocarcinoma cell line, registered under ATCC number ATCC-CRL-1469.

[0248] Initially, PANC1 cells were incubated for 48h in presence or absence of 50ug / ml conjugated ApoA-I antibody or anti-HEL-pHAb (isotype control) in cell culture media containing 10% normal human serum. Cells were released from tissue culture wells with 5mM EDTA / PBS solution, 10 minutes at 37C. The cells were then spun down and suspended into FACS buffer for analysis with Beckmann Coulter CytoFlex flow cytometer. For each condition, 10000 events were recorded and live cells were gated out. Live cell populations are shown in FIGS 3A-3B.

[0249] Surprisingly, more than 71% of cells internalized the conjugated antibody as compared to an isotype antibody control (FIG.3A), and more than 97% internalized as compared to an untreated control (FIG.3B). This showed that molecular hooks can efficiently be delivered to target cells on HDL molecules.

[0250] Time course study. PANC1 cells were incubated for 24, 48 or 96 hours in presence or absence of 50ug / ml conjugated ApoA-I antibody or anti-HEL-pHAb (isotype control) in cell culture media containing 10% normal human serum. As a positive control, PANC1 cells were incubated for 48 hours in the presence of conjugated ApoA-I antibody or anti- HEL- pHAb (isotype control) and PEI. Cells were released from tissue culture wells with 5mM EDTA / PBS solution, 10 minutes at 37C. The cells were then spun down and suspended into FACS buffer for analysis with Beckmann Coulter CytoFlex flow cytometer. For each condition, 10000 events were recorded and live cells were gated out. Live cell populations are shown in FIG.3C.

[0251] FIG.3C shows that internalization of the conjugated ApoA-I antibody increases over time and is durable until 96 hours (almost 70% internalization). The HEL–pHAb antibody used as a negative control exhibited minimal cellular uptake—only 1.15% at 48 hours and 58 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 1.09% at 96 hours—demonstrating no significant internalization. In contrast, the conjugated ApoA-I antibody, which constitutes the delivery vehicle for mRNA in COMED’s Molecular Hook system, demonstrated substantially higher intracellular uptake: 50.7% at 48 hours and 67.3% at 96 hours (FIG.3C, FIG.3D). These findings clearly indicate that COMED’s Molecular Hook technology enables efficient and progressive cellular internalization of the antibody conjugate over time.

[0252] Molecular Hook Antibodies are not Trapped in Endosomes once Internalized. One potential pitfall of cell internalization is endosomal entrapment. If an internalized antibody is trapped in an endosome, it is not available to bind its target and is eventually degraded.

[0253] pHAb is a fluorescent dye that binds to antibodies and is ideally suited for detecting antibodies that have been internalized by the cell. Since the pHAb dye is sensitive to intracellular pH, it emits fluorescence depending on whether the internalized antibody has undergone lysosomal degradation: a strong fluorescent signal is observed when the antibody is trapped in lysosomes. A weak signal is observed when the antibody remains in the cytoplasm, indicating successful evasion of lysosomal entrapment.

[0254] Applicant conducted imaging experiments (in combination with FACS) using pHAb and showed that the internalized antibody is not trapped in endosomes, but is readily available in the cell cytoplasm even after 93 hours (FIGS.4A-4B).

[0255] These experiments show that the disclosed molecular hooks can effectively internalize into target cells via HDL-mediated delivery and are able to deliver their load into the cytoplasm. The signal is still detectable 24 hours post-treatment, indicating intracellular persistence of the compound. The pHAb signal is mild, suggesting the compound has avoided lysosomal entrapment and is likely localized within the cytoplasm rather than degraded. Molecular hooks’ ability to avoid endosomal traps is important for their efficacy. The uptake rate within the first 24 hours is quantified as 48.8%, demonstrating efficient internalization. Additional Embodiments

[0256] Embodiment 1. An in vivo method for delivering ribonucleic acid (RNA) to cells of a subject in need thereof comprising administering to the subject (i) an RNA-HDL complex wherein the RNA is tethered to the exterior of a high-density lipoprotein (HDL), or 59 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 (ii) an RNA-molecular hook complex wherein the molecular hook is capable of tethering to the exterior of any HDL molecule.

[0257] Embodiment 2. A method for delivering ribonucleic acid (RNA) to a cell comprising contacting the cells with an RNA-HDL complex wherein the RNA is tethered to the exterior of an HDL, optionally wherein the RNA is tethered to the exterior of an HDL through a molecular hook.

[0258] Embodiment 3. The method of embodiment 2, wherein the contacting is performed in vitro or in vivo.

[0259] Embodiment 4. The method of embodiment 1, wherein the cells of the subject are behind the blood brain barrier of the subject.

[0260] Embodiment 5. The method of embodiment 4, wherein the cells of the subject are in the brain or the spine of the subject.

[0261] Embodiment 6. The method of any one of embodiments 1-5, wherein the molecular hook comprises: a first antigen binding domain that specifically binds to an exterior facing protein of the HDL; and a second antigen binding domain that specifically binds to the ribonucleic acid (RNA).

[0262] Embodiment 7. The method of embodiment 6, wherein the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C-II (Apo C-II).

[0263] Embodiment 8. The method of embodiment 6 or embodiment 7, wherein the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR- H2 and CDR-H3 of a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or accession No. NITE BP-02443, or CDR-L1, CDR-L2, CDR-L3, CDR- H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 , or 60 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO: 68.

[0264] Embodiment 9. The method of embodiment 6 or embodiment 7, wherein the first antigen binding domain specifically binds ApoA-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR- H2 and CDR-H3 of the EPR2913 monoclonal antibody, or amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

[0265] Embodiment 10. The method of embodiment 6 or embodiment 7, wherein the first antigen binding domain specifically binds Apo E and comprises an amino acid sequence selected from: (i) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of MAB41445 monoclonal antibody; (ii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42; (iii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44; (iv) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45 and CDR-H1, CDR-H2 and CDR-H3 61 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46; (v) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48; (vi) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 41 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 42; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 44; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 45 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 46; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 47 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 48; or (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 49 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 50.

[0266] Embodiment 11. The method of embodiment 6 or embodiment 7, wherein the first antigen binding domain specifically binds Apo C-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR- H2 and CDR-H3 of the 3E4 monoclonal antibody.

[0267] Embodiment 12. The method of any one of embodiments 6-11, wherein the second antigen binding domain that specifically binds to RNA comprises HuR RNA binding protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 1.

[0268] Embodiment 13. The method of any one of embodiments 6-11, wherein the second antigen binding domain that specifically binds to RNA comprises CDR-L1, CDR-L2, 62 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

[0269] Embodiment 14. The method of embodiment any one of embodiments 6-13, wherein the first antigen binding domain and or the second antigen binding domain comprises: a single-chain variable fragment (scFv), a tandem scFv dimer [(scFv)₂], an scFv- Fc fusion, a Fab, a Fab′, an F(ab′)₂ fragment, a bispecific antibody, a multispecific antibody, a diabody, or Fc-fusion proteins, as well as isotypes, engineered variants, or functional fragments thereof derived from such formats.

[0270] Embodiment 15. The method of any one of embodiments 1-14, wherein the molecular hook comprises a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker.

[0271] Embodiment 16. The method of any one of embodiments 1-15, wherein the RNA comprises mRNA, and optionally wherein the mRNA encodes a therapeutic protein; and optionally wherein the mRNA encodes a vaccine.

[0272] Embodiment 17. The method of any one of embodiments 1-8, wherein the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain of the anti-APOA-I binding antibody and the HuR RNA binding protein are linked together by a flexible linker selected from SEQ ID NOs: 76-79.

[0273] Embodiment 18. The method of any one of embodiments 1-8 or 17, wherein the molecular hook comprises SEQ ID NO: 75 and SEQ ID NO: 67.

[0274] Embodiment 19. A molecular hook comprising: a first antigen binding domain that specifically binds to an exterior facing protein of a high density lipoprotein (HDL); and a second antigen binding domain that specifically binds to a ribonucleic acid (RNA). 63 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0275] Embodiment 20. The molecular hook of embodiment 19, wherein the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C (Apo C).

[0276] Embodiment 21. The molecular hook of embodiment 19 or embodiment 20, wherein the first antigen binding domain specifically binds ApoA-I and comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 comprising amino acid sequences that are respectively same as amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of monoclonal antibody produced from a hybridoma of accession No. NITE BP- 02442 or accession No. NITE BP-02443, or CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 , or a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO: 68.

[0277] Embodiment 22. The molecular hook of embodiment 19 or embodiment 20, wherein the first antigen binding domain specifically binds ApoA-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the EPR2913 monoclonal antibody, or amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

[0278] Embodiment 23. The molecular hook of embodiment 19 or embodiment 20, wherein the first antigen binding domain specifically binds Apo E and comprises an amino acid sequence selected from: 64 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 (i) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of MAB41445 monoclonal antibody; (ii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42; (iii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44; (iv) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46; (v) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48; (vi) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 41 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 42; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 44; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 45 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 46; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 47 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 48; or 65 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 49 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 50.

[0279] Embodiment 24. The molecular hook of embodiment 19 or embodiment 20, wherein the first antigen binding domain specifically binds Apo C and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the 3E4 monoclonal antibody.

[0280] Embodiment 25. The molecular hook of any one of embodiments 19-24, wherein the second antigen binding domain that specifically binds to RNA comprises HuR RNA binding protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 1.

[0281] Embodiment 26. The molecular hook of any one of embodiments 19-24, wherein the second antigen binding domain that specifically binds to RNA comprises CDR-L1, CDR- L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

[0282] Embodiment 27. The molecular hook of any one of embodiments 19-26, wherein the first antigen binding domain and or the second antigen binding domain comprises single- chain variable fragment (scFv), a tandem scFv dimer [(scFv)₂], an scFv-Fc fusion, a Fab, a Fab′, an F(ab′)₂ fragment, a bispecific antibody, a multispecific antibody, a diabody, or Fc- fusion proteins, as well as isotypes, engineered variants, or functional fragments thereof derived from such formats.

[0283] Embodiment 28. The molecular hook of any one of embodiments 19-27, wherein the molecular hook comprises a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker. 66 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0284] Embodiment 29. The molecular hook of any one of embodiments 19-28, wherein the RNA is mRNA; optionally wherein the mRNA encodes a therapeutic protein or a vaccine.

[0285] Embodiment 30. The molecular hook of any one of embodiments 19-21, wherein the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain of the anti-APOA-I binding antibody and the HuR RNA binding protein are linked together by a flexible linker selected from SEQ ID NOs: 76- 79.

[0286] Embodiment 31. The molecular hook of any one of embodiments 19-21 or 30, wherein the molecular hook comprises SEQ ID NO: 75 and SEQ ID NO: 67.

[0287] Embodiment 32. Use of an RNA-HDL complex for the manufacture of a medicament for delivering ribonucleic acid (RNA) to cells, wherein the RNA-HDL complex comprises an RNA tethered to the exterior of a high-density lipoprotein (HDL), optionally through a molecular hook.

[0288] Embodiment 33. Use of an RNA-molecular hook complex for the manufacture of a medicament for delivering ribonucleic acid (RNA) to cells.

[0289] Embodiment 34. The use of embodiment 32 or embodiment 33, wherein the molecular hook comprises: a first antigen binding domain that specifically binds to an exterior facing protein of the HDL; and a second antigen binding domain that specifically binds to the ribonucleic acid (RNA).

[0290] Embodiment 35. The use of embodiment 34, wherein the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C-II (Apo C-II).

[0291] Embodiment 36. The use of embodiment 34 or embodiment 35, wherein the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR- L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR- H3 of a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 67 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 or accession No. NITE BP-02443, or CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 , or a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO: 68.

[0292] Embodiment 37. The use of embodiment 34 or embodiment 35, wherein the first antigen binding domain specifically binds ApoA-II and comprises CDR-L1, CDR-L2, CDR- L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR- H3 of the EPR2913 monoclonal antibody, or amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

[0293] Embodiment 38. The use of embodiment 34 or embodiment 35, wherein the first antigen binding domain specifically binds Apo E and comprises an amino acid sequence selected from: (i) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of MAB41445 monoclonal antibody; (ii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42; 68 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 (iii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44; (iv) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46; (v) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48; (vi) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 41 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 42; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 44; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 45 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 46; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 47 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 48; or (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 49 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 50.

[0294] Embodiment 39. The use of embodiment 34 or embodiment 35, wherein the first antigen binding domain specifically binds Apo C-II and comprises CDR-L1, CDR-L2, CDR- L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR- H3 of the 3E4 monoclonal antibody. 69 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0295] Embodiment 40. The use of any one of embodiments 34-39, wherein the second antigen binding domain that specifically binds to RNA comprises HuR RNA binding protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 1.

[0296] Embodiment 41. The use of any one of embodiments 34-39, wherein the second antigen binding domain that specifically binds to RNA comprises CDR-L1, CDR-L2, CDR- L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

[0297] Embodiment 42. The use of any one of embodiments 34-41, wherein the first antigen binding domain and or the second antigen binding domain comprises a single-chain variable fragment (scFv), a tandem scFv dimer [(scFv)₂], an scFv-Fc fusion, a Fab, a Fab′, an F(ab′)₂ fragment, a bispecific antibody, a multispecific antibody, a diabody, or Fc-fusion proteins, as well as isotypes, engineered variants, or functional fragments thereof derived from such formats.

[0298] Embodiment 43. The use of any one of embodiments 33-42, wherein the molecular hook comprises a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker.

[0299] Embodiment 44. The use of any one of embodiments 32-43, wherein the RNA comprises mRNA, and optionally wherein the mRNA encodes a therapeutic protein; and optionally wherein the mRNA encodes a vaccine.

[0300] Embodiment 45. The use of any one of embodiments 32-36, wherein the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain of the anti-APOA-I binding antibody and the HuR RNA binding protein are linked together by a flexible linker selected from SEQ ID NOs: 76-79. 70 4865-1048-3383.1Atty. Dkt. No.: 139886-1010

[0301] Embodiment 46. The use of any one of embodiments 32-36 or 45, wherein the molecular hook comprises SEQ ID NO: 75 and SEQ ID NO: 67. List of Sequences SEQ ID NO: 1, HuR RNA binding protein, Homo sapiens MSNGYEDHMAEDCRGDIGRTNLIVNYLPQNMTQDELRSLFSSIGEVESAKLIRDKVA GHSLGYGFVNYVTAKDAERAINTLNGLRLQSKTIKVSYARPSSEVIKDANLYISGLPR TMTQKDVEDMFSRFGRIINSRVLVDQTTGLSRGVAFIRFDKRSEAEEAITSFNGHKPP GSSEPIAVKFAANPNQNKNVALLSQLYHSPARRFGGPVHHQAQRFRFSPMGVDHMS GLSGVNVPGNASSGWCIFIYNLGQDADEGILWQMFGPFGAVTNVKVIRDFNTNKCK GFGFVTMTNYEEAAMAIASLNGYRLGDKILQVSFKTNKSHK SEQ ID NO: 2, CDR-L1 of an RNA antibody, Artificial sequence SVSS SEQ ID NO: 3, CDR-L2 of an RNA antibody, Artificial sequence YSASSLY SEQ ID NO: 4, CDR-L3 of an RNA antibody, Artificial sequence SYSYPY SEQ ID NO: 5, CDH-H1 of an RNA antibody, Artificial sequence SISSSYIH SEQ ID NO: 6, CDH-H2 of an RNA antibody, Artificial sequence YIYPYYGSTY SEQ ID NO: 7, CDR-H3 of an RNA antibody, Artificial sequence YYSSGGSYYRYSRGFD SEQ ID NO: 8, CDR-L1 of an RNA antibody, Artificial sequence SVYY SEQ ID NO: 9, CDR-L3 of an RNA antibody, Artificial sequence FLSSLI SEQ ID NO: 10, CDR-H1 of an RNA antibody, Artificial sequence YLYSSSSMH SEQ ID NO: 11, CDR-H2 of an RNA antibody, Artificial sequence SIYPYSGSTS SEQ ID NO: 12, CDR-H3 of an RNA antibody, Artificial sequence YWSYKYGRAMD SEQ ID NO: 13, CDR-L1 of an RNA antibody, Artificial sequence YVYS SEQ ID NO: 14, CDR-L3 of an RNA antibody, Artificial sequence SYRHLF 71 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 SEQ ID NO: 15, CDR-H1 of an RNA antibody, Artificial sequence SLSSYYSMH SEQ ID NO: 16, CDR-H3 of an RNA antibody, Artificial sequence SKKYAYKHAMD SEQ ID NO: 17, CDR-L1 of an APOA-II antibody, Artificial sequence RSSKSLLYKDGKTYLN SEQ ID NO: 18, CDR-L2 of an APOA-II antibody, Artificial sequence LMSTRAS SEQ ID NO: 19, CDR-L3 of an APOA-II antibody, Artificial sequence QQLVEYPLT SEQ ID NO: 20, CDR-H1 of an APOA-II antibody, Artificial sequence GYTFTNYWMH SEQ ID NO: 21, CDR-H2 of an APOA-II antibody, Artificial sequence NIYPGSGNTNYNEKFK SEQ ID NO: 22, CDR-H3 of an APOA-II antibody, Artificial sequence RYGYVDWFAY SEQ ID NO: 23, CDR-L1 of an APOA-II antibody, Artificial sequence RASQDTSNYLN SEQ ID NO: 24, CDR-L2 of an APOA-II antibody, Artificial sequence YTSRLHS SEQ ID NO: 25, CDR-L3 of an APOA-II antibody, Artificial sequence QQGNTLPYT SEQ ID NO: 26, CDR-H1 of an APOA-II antibody, Artificial sequence GYTFTSYWMH SEQ ID NO: 27, CDR-H2 of an APOA-II antibody, Artificial sequence FINPSTGYTENNQRFN SEQ ID NO: 28, CDR-H3 of an APOA-II antibody, Artificial sequence RPYNPYAMDY SEQ ID NO: 29, CDR-L1 of an APOA-II antibody, Artificial sequence RASSSLSSSYLH SEQ ID NO: 30, CDR-L2 of an APOA-II antibody, Artificial sequence STSNLAS SEQ ID NO: 31, CDR-L3 of an APOA-II antibody, Artificial sequence QQFSVFPLT SEQ ID NO: 32, CDR-H1 of an APOA-II antibody, Artificial sequence 72 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 NYGMN SEQ ID NO: 33, CDR-H2 of an APOA-II antibody, Artificial sequence WKNTYTGESTYADDFK SEQ ID NO: 34, CDR-H3 of an APOA-II antibody, Artificial sequence RDGSKYKIFDY SEQ ID NO: 35, CDR-L1 of an APOA-II antibody, Artificial sequence RASQDISNYLN SEQ ID NO: 36, CDR-L2 of an APOA-II antibody, Artificial sequence YTSRLQS SEQ ID NO: 37, CDR-L3 of an APOA-II antibody, Artificial sequence QQGNTLPYT SEQ ID NO: 38, CDR-H1 of an APOA-II antibody, Artificial sequence GYTFTSYWMH SEQ ID NO: 39, CDR-H2 of an APOA-II antibody, Artificial sequence FINPSTGYTENNQNFK SEQ ID NO: 40, CDR-H3 of an APOA-II antibody, Artificial sequence RTYNPYGMDY SEQ ID NO: 41, light chain variable region of an ApoE antibody, Artificial sequence DVLMTQTPLSLPVSLGDQASISCRSSQSIVHSNGNTYLEWYLQKPGQSPKLLIYKVSN RFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPYTFGGGTKLEIKR SEQ ID NO: 42, heavy chain variable region of an ApoE antibody, Artificial sequence RVQLQQSGPGLVKPGASVKISCKASGYTFTDYYINWVRQRPGQGLEWIGWIFPGSGI TYYNEKFKGKATLTVDRSSSTAYMLLSSLTSEDSAVYFCARYAYGSPFAYWGQGTL VTVSA SEQ ID NO: 43, light chain variable region of an ApoE antibody, Artificial sequence DVLMTQTPLSLPVSLGDQASISCRSSQNIIHSNGNTYLEWFLQRPGQSPELLIYKVSNR FSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPYTFGGGTKLEIKR SEQ ID NO: 44, heavy chain variable region of an ApoE antibody, Artificial sequence QVQLQQSGPELVKPGASVKISCKASGYIFTDYYINWVKRRPGQGLEWIGWIFPGSGV SYYNEQFKGKATLTVDKPSNTAYIFLSRLTSEDSAVYFCARYYSSSPFAYWGQGTLV TVSA SEQ ID NO: 45, light chain variable region of an ApoE antibody, Artificial sequence DVLMTQTPLSLPVSLGDQASISCRSSQNIVYSNGNTYLEWYLQKPGQSPKLLIYKVSN RFSGVPDRFSGSGSGTYFTLKISRVEAEDLGIYYCFQGSHVPYTFGGGTKLEIKR SEQ ID NO: 46, heavy chain variable region of an ApoE antibody, Artificial sequence QVQLQQSGPDLVKPGASVKISCKASGYTFIDYYINWAKQRPGQGLEWIGWIFPGSGS TYYNEKFKGKATLTVDKSSSTAYMFLSSLTSEDSAVFFCARYYGSSPFAYWGQGTL VTVSA 73 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 SEQ ID NO: 47, light chain variable region of an ApoE antibody, Artificial sequence DVLMTQTPLSLPVSLGDQASISCRSSQNIIHSNGNTYLEWFLQKPGQSPKLLIYKVSN RFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPYTFGGGTKLEIKR SEQ ID NO: 48, heavy chain variable region of an ApoE antibody, Artificial sequence QIQLQQSGPELVKPGASVKISCKASGYTFTDYYINWVRQRPGQGLEWIGWIYPESFN TYYNEKFKGKATLTVDTSSNTAYMQLSSLTSEDSAVYFCARYYVSSPFAYWGQGTL VTVSA SEQ ID NO: 49, light chain variable region of an ApoE antibody, Artificial sequence DVLMTQSPLSLPVSLGDQASISCRSSQSIVHRNGNTYLEWYLQKPGQSPKLLIYKVSN RFSGVPDRFSGSGSGTDFTLKISRVEAEDLGVYYCFQGSHVPYTFGGGTKLEIRR SEQ ID NO: 50, heavy chain variable region of an ApoE antibody, Artificial sequence QVQLQQSGPELVKPGASVKISCKASGYTFSDYYINWVKQRPGQGLEWIGWIFPGSGS SYYNEKFKGKATLTVDKSSRTAYMLLSSLTSEDSAVYFCARYYGSSPFAYWGQGTL VTVST SEQ ID NO: 51, CDR-L1 of the light chain variable region of SEQ ID NOS: 41, 43, 45 and 47 RSSQSIVHSNGNTYLE SEQ ID NO: 52, CDR-L2 of the light chain variable region of SEQ ID NOS: 41, 43, 45 and 47 KVSNRFS SEQ ID NO: 53, CDR-L3 of the light chain variable region of SEQ ID NOS: 41, 43, 45 and 47 FQGSHVPYT SEQ ID NO: 54, CDR-H1 of the heavy chain variable region of SEQ ID NOS: 42, 44, 46, 48 and 50 DYYIN SEQ ID NO: 55, CDR-H2 of the heavy chain variable region of SEQ ID NO: 42 WIFPGSGITYYNEKFKG SEQ ID NO: 56, CDR-H3 of the heavy chain variable region of SEQ ID NO: 42 YAYGSPFAY SEQ ID NO: 57, CDR-L1 of the light chain variable region of SEQ ID NOS: 43, 45, 47, and 49 RSSQNIIHSNGNTYLE SEQ ID NO: 58, CDR-H2 of the heavy chain variable region of SEQ ID NO: 44 WIFPGSGVSYYNEQFKG SEQ ID NO: 59, CDR-H3 of the heavy chain variable region of SEQ ID NO: 44 YYSSSPFAY SEQ ID NO: 60, CDR-L1 of the light chain variable region of SEQ ID NO: 45 74 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 RSSQNIVYSNGNTYLE SEQ ID NO: 61, CDR-H2 of the heavy chain variable region of SEQ ID NO: 46 WIFPGSGSTYYNEKFKG SEQ ID NO: 62, CDR-H3 of the heavy chain variable region of SEQ ID NOS: 46 and 50 YYGSSPFAY SEQ ID NO: 63, CDR-H2 of the heavy chain variable region of SEQ ID NO: 48 WIYPESFNTYYNEKFKG SEQ ID NO: 64, CDR-H3 of the heavy chain variable region of SEQ ID NO: 48 YYVSSPFAY SEQ ID NO: 65, CDR-L1 of the light chain variable region of SEQ ID NO: 49 RSSQSIVHRNGNTYLE SEQ ID NO: 66, CDR-H1 of the heavy chain variable region of SEQ ID NO: 50 WIFPGSGSSYYNEKFKG SEQ ID NO: 67, Anti-APOA-I light chain IVMTQTPSSKSVPVGGTVTINCQASESVSSNNYLSWFQQKPGQPPKLLIYLASTLVSG VPSRFKGSGSGTQFTLTISDVVCDDAATYYCAGYKKRSTDVIAFGGGTEVVVKGDP VAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTVTWEVDGTTQTTGIENSKTPQNS ADCTYNLSSTLTLTSTQYNSHKEYTCKVTQGTTSVVQSFNRGDC SEQ ID NO: 68, Anti-APOA-I heavy chain QEQLVESGGGLVQPEGSLALTCTASGFSFSTNYICWVRQAPGKGLEWVGCIDNGDA STYYASWAKGRFTISKTSSTTLTLQMTSLTAADTATYFCARYLAFDLWGPGTLVTVS SGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSV RQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPELLGGP SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQF NSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARGQPLEPKVYTMGPP REELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSV PTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGK SEQ ID NO: 69, CDR-L1 Anti-APOA-I light chain QASESVSSNNYLS SEQ ID NO: 70, CDR-L2 Anti-APOA-I light chain LASTLVS SEQ ID NO: 71, CDR-L3 Anti-APOA-Ilight chain AGYKKRSTDVIA SEQ ID NO: 72, CDR-H1 Anti-APOA-I heavy chain GFSFSTN SEQ ID NO: 73, CDR-H2 Anti-APOA-I heavy chain DNGDAS SEQ ID NO: 74, CDR-H3 Anti-APOA-I heavy chain 75 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 YLAFDL SEQ ID NO: 75, Molecular Hook Heavy chain (ApoA-I-Linker-HuR RNA binding protein) QEQLVESGGGLVQPEGSLALTCTASGFSFSTNYICWVRQAPGKGLEWVGCIDNGDA STYYASWAKGRFTISKTSSTTLTLQMTSLTAADTATYFCARYLAFDLWGPGTLVTVS SGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTVTWNSGTLTNGVRTFPSV RQSSGLYSLSSVVSVTSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPELLGGP SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLREQQF NSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPAPIEKTISKARGQPLEPKVYTMGPP REELSSRSVSLTCMINGFYPSDISVEWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSV PTSEWQRGDVFTCSVMHEALHNHYTQKSISRSPGKGGGGSGGGGSGGGGSGGGGSS NGYEDHMAEDCRGDIGRTNLIVNYLPQNMTQDELRSLFSSIGEVESAKLIRDKVAGH SLGYGFVNYVTAKDAERAINTLNGLRLQSKTIKVSYARPSSEVIKDANLYISGLPRTM TQKDVEDMFSRFGRIINSRVLVDQTTGLSRGVAFIRFDKRSEAEEAITSFNGHKPPGSS EPITVKFAANPNQNKNVALLSQLYHSPARRFGGPVHHQAQRFRFSPMGVDHMSGLS GVNVPGNASSGWCIFIYNLGQDADEGILWQMFGPFGAVTNVKVIRDFNTNKCKGFG FVTMTNYEEAAMAIASLNGYRLGDKILQVSFKTNKSHK The Light chain (ApoA-I- HuR RNA binding protein) for this molecular hook comprises SEQ ID NO: 67 (the light chain of the APOA-I antibody). SEQ ID NO: 76, Linker GGGGSGGGGS SEQ ID NO: 77, Linker GGGGSGGGGSGGGGS SEQ ID NO: 78, Linker GGGGSGGGGSGGGGSGGGGS SEQ ID NO: 79, Linker GGSGGGGSGGGGSGG Equivalents

[0302] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this technology belongs.

[0303] The present technology illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown 76 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 and described or portions thereof, but it is recognized that various modifications are possible within the scope of the present technology claimed.

[0304] Thus, it should be understood that the materials, methods, and examples provided here are representative of preferred aspects, are exemplary, and are not intended as limitations on the scope of the present technology.

[0305] It should be understood that although the present invention has been specifically disclosed by certain aspects, embodiments, and optional features, modification, improvement and variation of such aspects, embodiments, and optional features can be resorted to by those skilled in the art, and that such modifications, improvements and variations are considered to be within the scope of this disclosure.

[0306] The present technology has been described broadly and generically herein. Each of the narrower species and sub-generic groupings falling within the generic disclosure also form part of the present technology. This includes the generic description of the present technology with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

[0307] In addition, where features or aspects of the present technology are described in terms of Markush groups, those skilled in the art will recognize that the present technology is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0308] All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety, to the same extent as if each were incorporated by reference individually. In case of conflict, the present specification, including definitions, will control.

[0309] Other aspects are set forth within the following claims. 77 4865-1048-3383.1

Claims

Atty. Dkt. No.: 139886-1010 WHAT IS CLAIMED IS:

1. An in vivo method for delivering ribonucleic acid (RNA) to cells of a subject in need thereof comprising administering to the subject (i) an RNA-HDL complex wherein the RNA is tethered to the exterior of a high-density lipoprotein (HDL), or (ii) an RNA-molecular hook complex wherein the molecular hook is capable of tethering to the exterior of any HDL molecule.

2. A method for delivering ribonucleic acid (RNA) to a cell comprising contacting the cells with an RNA-HDL complex wherein the RNA is tethered to the exterior of an HDL, optionally wherein the RNA is tethered to the exterior of an HDL through a molecular hook.

3. The method of claim 2, wherein the contacting is performed in vitro or in vivo.

4. The method of claim 1, wherein the cells of the subject are behind the blood brain barrier of the subject.

5. The method of claim 4, wherein the cells of the subject are in the brain or the spine of the subject.

6. The method of any one of claims 1-5, wherein the molecular hook comprises: a first antigen binding domain that specifically binds to an exterior facing protein of the HDL; and a second antigen binding domain that specifically binds to the ribonucleic acid (RNA).

7. The method of claim 6, wherein the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C-II (Apo C-II).

8. The method of claim 6 or claim 7, wherein the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or accession No. NITE BP-02443, or CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 , or 78 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO:

68.

9. The method of claim 6 or claim 7, wherein the first antigen binding domain specifically binds ApoA-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the EPR2913 monoclonal antibody, or amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

10. The method of claim 6 or claim 7, wherein the first antigen binding domain specifically binds Apo E and comprises an amino acid sequence selected from: (i) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of MAB41445 monoclonal antibody; (ii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42; (iii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44; (iv) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46; (v) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48; 79 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 (vi) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 41 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 42; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 44; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 45 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 46; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 47 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 48; or (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 49 and a heavy chain variable region having at least 90% identity to SEQ ID NO:

50.

11. The method of claim 6 or claim 7, wherein the first antigen binding domain specifically binds Apo C-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the 3E4 monoclonal antibody.

12. The method of any one of claims 6-11, wherein the second antigen binding domain that specifically binds to RNA comprises HuR RNA binding protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:

1.

13. The method of any one of claims 6-11, wherein the second antigen binding domain that specifically binds to RNA comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

14. The method of claim any one of claims 6-13, wherein the first antigen binding domain and or the second antigen binding domain comprises an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody. 80 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 15. The method of any one of claims 1-14, wherein the molecular hook comprises a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker.

16. The method of any one of claims 1-15, wherein the RNA comprises mRNA, and optionally wherein the mRNA encodes a therapeutic protein; and optionally wherein the mRNA encodes a vaccine.

17. The method of any one of claims 1-8, wherein the molecular hook comprises an anti- APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain of the anti-APOA-I binding antibody and the HuR RNA binding protein are linked together by a flexible linker selected from SEQ ID NOs: 76-79.

18. The method of any one of claims 1-8 or 17, wherein the molecular hook comprises SEQ ID NO: 75 and SEQ ID NO:

67.

19. A molecular hook comprising: a first antigen binding domain that specifically binds to an exterior facing protein of a high density lipoprotein (HDL); and a second antigen binding domain that specifically binds to a ribonucleic acid (RNA).

20. The molecular hook of claim 19, wherein the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C (Apo C).

21. The molecular hook of claim 19 or claim 20, wherein the first antigen binding domain specifically binds ApoA-I and comprises HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 comprising amino acid sequences that are respectively same as amino acid sequences of HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 of monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or accession No. NITE BP- 02443, or CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 , or 81 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO:

68.

22. The molecular hook of claim 19 or claim 20, wherein the first antigen binding domain specifically binds ApoA-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the EPR2913 monoclonal antibody, or amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

23. The molecular hook of claim 19 or claim 20, wherein the first antigen binding domain specifically binds Apo E and comprises an amino acid sequence selected from: (i) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of MAB41445 monoclonal antibody; (ii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42; (iii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44; (iv) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46; (v) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48; 82 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 (vi) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 41 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 42; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 44; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 45 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 46; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 47 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 48; or (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 49 and a heavy chain variable region having at least 90% identity to SEQ ID NO:

50.

24. The molecular hook of claim 19 or claim 20, wherein the first antigen binding domain specifically binds Apo C and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the 3E4 monoclonal antibody.

25. The molecular hook of any one of claims 19-24, wherein the second antigen binding domain that specifically binds to RNA comprises HuR RNA binding protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:

1.

26. The molecular hook of any one of claims 19-24, wherein the second antigen binding domain that specifically binds to RNA comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 . 83 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 27. The molecular hook of any one of claims 19-26, wherein the first antigen binding domain and or the second antigen binding domain comprises an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody.

28. The molecular hook of any one of claims 19-27, wherein the molecular hook comprises a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker.

29. The molecular hook of any one of claims 19-28, wherein the RNA is mRNA; optionally wherein the mRNA encodes a therapeutic protein or a vaccine.

30. The molecular hook of any one of claims 19-21, wherein the molecular hook comprises an anti-APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain of the anti-APOA-I binding antibody and the HuR RNA binding protein are linked together by a flexible linker selected from SEQ ID NOs: 76-79.

31. The molecular hook of any one of claims 19-21 or 30, wherein the molecular hook comprises SEQ ID NO: 75 and SEQ ID NO:

67.

32. Use of an RNA-HDL complex for the manufacture of a medicament for delivering ribonucleic acid (RNA) to cells, wherein the RNA-HDL complex comprises an RNA tethered to the exterior of a high-density lipoprotein (HDL), optionally through a molecular hook.

33. Use of an RNA-molecular hook complex for the manufacture of a medicament for delivering ribonucleic acid (RNA) to cells.

34. The use of claim 32 or claim 33, wherein the molecular hook comprises: a first antigen binding domain that specifically binds to an exterior facing protein of the HDL; and a second antigen binding domain that specifically binds to the ribonucleic acid (RNA).

35. The use of claim 34, wherein the exterior facing protein is selected from the group consisting of Apolipoprotein A-I (ApoA-I), Apolipoprotein A-II (ApoA-II), Apolipoprotein E (Apo E) and Apolipoprotein C-II (Apo C-II).

36. The use of claim 34 or claim 35, wherein the first antigen binding domain specifically binds ApoA-I and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 84 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of a monoclonal antibody produced from a hybridoma of accession No. NITE BP-02442 or accession No. NITE BP- 02443, or CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences from a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 69 SEQ ID NO: 70 SEQ ID NO: 71 SEQ ID NO: 72 SEQ ID NO: 73 SEQ ID NO: 74 , or a light chain variable region as shown in SEQ ID NO: 67 and a heavy chain variable region as shown in SEQ ID NO:

68.

37. The use of claim 34 or claim 35, wherein the first antigen binding domain specifically binds ApoA-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the EPR2913 monoclonal antibody, or amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 17 SEQ ID NO: 18 SEQ ID NO: 19 SEQ ID NO: 20 SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23 SEQ ID NO: 24 SEQ ID NO: 25 SEQ ID NO: 26 SEQ ID NO: 27 SEQ ID NO: 28 SEQ ID NO: 29 SEQ ID NO: 30 SEQ ID NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 SEQ ID NO: 37 SEQ ID NO: 38 SEQ ID NO: 39 SEQ ID NO: 40 .

38. The use of claim 34 or claim 35, wherein the first antigen binding domain specifically binds Apo E and comprises an amino acid sequence selected from: (i) CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as the amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of MAB41445 monoclonal antibody; (ii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 41 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 42; (iii) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 43 and CDR-H1, CDR-H2 and CDR-H3 85 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 44; (iv) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 45 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 46; (v) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 47 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 48; (vi) CDR-L1, CDR-L2, and CDR-L3 that are respectively same as amino acid sequences of CDR-L1, CDR-L2, and CDR-L3 of SEQ ID NO: 49 and CDR-H1, CDR-H2 and CDR-H3 that are respectively same as amino acid sequences of CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 50; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 41 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 42; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 43 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 44; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 45 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 46; (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 47 and a heavy chain variable region having at least 90% identity to SEQ ID NO: 48; or (vii) a light chain variable region having at least 90% identity to SEQ ID NO: 49 and a heavy chain variable region having at least 90% identity to SEQ ID NO:

50.

39. The use of claim 34 or claim 35, wherein the first antigen binding domain specifically binds Apo C-II and comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences that are respectively same as amino acid sequences of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of the 3E4 monoclonal antibody.

40. The use of any one of claims 34-39, wherein the second antigen binding domain that specifically binds to RNA comprises HuR RNA binding protein comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:

1. 86 4865-1048-3383.1Atty. Dkt. No.: 139886-1010 41. The use of any one of claims 34-39, wherein the second antigen binding domain that specifically binds to RNA comprises CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 comprising amino acid sequences of a single line of the following table: CDR-L1 CDR-L2 CDR-L3 CDR-H1 CDR-H2 CDR-H3 SEQ ID NO: 2 SEQ ID NO: 3 SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID NO: 3 SEQ ID NO: 9 SEQ ID NO: 10 SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13 SEQ ID NO: 3 SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 11 SEQ ID NO: 16 .

42. The use of any one of claims 34-41, wherein the first antigen binding domain and or the second antigen binding domain comprises an scFv, an (scFv)2, an scFvFc, a Fab, a Fab’, a F(ab’)2, a bispecific antibody or a diabody.

43. The use of any one of claims 33-42, wherein the molecular hook comprises a linker between the first antigen binding domain and the second antigen binding domain, optionally wherein the linker is an enzymatically cleavable linker, a hydrolysable linker, a pH sensitive linker, a photolabile linker, or a self immolative linker.

44. The use of any one of claims 32-43, wherein the RNA comprises mRNA, and optionally wherein the mRNA encodes a therapeutic protein; and optionally wherein the mRNA encodes a vaccine.

45. The use of any one of claims 32-36, wherein the molecular hook comprises an anti- APOA-I binding antibody having a heavy chain sequence having at least 90% identity to SEQ ID NO: 68, a light chain having at least 90% identity to SEQ ID NO: 67, and a HuR RNA binding protein having a sequence as shown by SEQ ID NO: 1, wherein the heavy chain of the anti-APOA-I binding antibody and the HuR RNA binding protein are linked together by a flexible linker selected from SEQ ID NOs: 76-79.

46. The use of any one of claims 32-36 or 45, wherein the molecular hook comprises SEQ ID NO: 75 and SEQ ID NO:

67. 87 4865-1048-3383.1

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