Lipid nanoparticle formulations containing human milk sugars

Abstract

Methods and materials for lipid nanoparticle delivery of nucleic acids are provided herein. For example, compositions containing human milk oligosaccharides in combination with lipid nanoparticles having nucleic acid cargoes are provided herein, as are methods that include administering, to mammals in need thereof, human milk oligosaccharides and lipid nanoparticles that carry nucleic acid cargoes.

Description

[0001]Attorney Docket No.26100-0153WO1 2023-273 LIPID NANOPARTICLE FORMULATIONS CONTAINING HUMAN MILK SUGARS CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority from U.S. Provisional Application Serial No. 63 / 522,441, filed June 22, 2023. The disclosure of the prior application is considered part of (and is incorporated by reference in) the disclosure of this application. STATEMENT AS TO FEDERALLY SPONSORED RESEARCH This invention was made with government support under HD098860 awarded by the National Institutes of Health. The government has certain rights in the invention. TECHNICAL FIELD This document relates to methods and materials for lipid nanoparticle delivery of nucleic acids. For example, this document relates to compositions containing human milk oligosaccharides in combination with lipid nanoparticles having nucleic acid cargoes. This document also relates to methods that include administering, to mammals in need thereof, human milk oligosaccharides and lipid nanoparticles that have nucleic acid cargoes. BACKGROUND RNA therapeutics have a broad range of applications for treating diseases in which one or more proteins is dysregulated (e.g., upregulated or downregulated). Lipid nanoparticles (LNPs) are efficient carriers for delivering nucleic acid cargo into cells, facilitating gene delivery, mRNA delivery, antisense, and RNA interference, among other uses. LPNs can serve as drug delivery vehicles for treating a broad range of genetic and acquired diseases. Notably, two COVID-19 vaccines utilized LNPs as their delivery system. However, challenges can arise when attempting nucleic acid delivery in the presence of inflammation. LNPs themselves can induce inflammation, and LNPs may be less effective in patients experiencing inflammatory conditions. Despite achieving some degree of success in the parenteral delivery of nucleic acids via LNPs, overcoming inflammation-associated barriers remains a significant hurdle in maximizing LNP therapeutic potential. Attorney Docket No.26100-0153WO1 2023-273 SUMMARY This document is based, at least in part, on the development of methods and materials that can be used to improve the efficacy of LNP delivery of therapeutic agents (e.g., RNAs). Inflammation often occurs in the body due to disease, and normal LNP mRNA delivery efficacy often is reduced with inflammation. The methods and materials provided herein can rescue this decrease in efficacy, and therefore can improve the effectiveness of LNP- mediated delivery of nucleic acids (e.g., RNA). In a first aspect, this document features a method for delivering a therapeutic agent to a subject. The method can include, or consist essentially of, administering to the subject a human milk oligosaccharide (HMO) and a lipid-containing particle containing the therapeutic agent. The HMO can include 3-Fucosyllactose, 6′Sialyllactose, 3′Sialyllactose (3′S), neutral penta-to-hexasaccharides, 2′Fucosyllactose, Sialylated Oligosaccharides, Lacto- N-fucopentaose I, Lacto-N-tetraose, and / or Neutral tetrasaccharides. In some cases, the HMO can be 3′S. The HMO can be administered intravenously, intramuscularly, or subcutaneously. The HMO can be administered at a dose of about 0.1 mg / kg to about 100 mg / kg, or at a dose of about 0.1 mg / kg to about 50 mg / kg. The lipid-containing particle can be a lipid nanoparticle. The lipid-containing particle can be administered intravenously, intramuscularly, or subcutaneously. The HMO and the lipid-containing particle can be administered simultaneously or can be administered sequentially. The therapeutic agent can be a nucleic acid. The nucleic acid can include an RNA (e.g., a mRNA, siRNA, shRNA, miRNA, antisense RNA, guide RNA, long non-coding RNA, transfer RNA, ribosomal RNA, dsRNA, or RNA aptamer). The RNA can be an mRNA encoding luciferase. The RNA can be administered at a dose of about 0.05 mg / kg to about 5 mg / kg. In some cases, the RNA can be administered at a dose of about 0.5 mg / kg. The lipid-containing particle can include an ionizable lipid, 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol or a derivative thereof, and a polyethylene glycol-lipid. The subject can be a healthy subject. The subject can have an inflammatory condition. The inflammatory condition can include inflammation in a lung, liver, spleen, blood vessel, brain, lymph node, or gastrointestinal tract. The inflammatory condition can have been induced by a stimulator such as a lipopolysaccharide, lipid-containing particle, infection, injury, toxin, chronic disease, smoking, or any Attorney Docket No.26100-0153WO1 2023-273 combination thereof. The stimulator can be LPS or lipid-containing particles. The subject can be a mammal (e.g., a mouse or a human). The subject can have been identified as having an infectious disease, autoimmune disease, cardiovascular disease, respiratory disease, neurological disease, cancer, metabolic disease, musculoskeletal disease, hematologic disease, or any combination thereof. In another aspect, this document features a composition that contains, consists essentially of, or consists of a human milk oligosaccharide (HMO) and a lipid-containing particle containing a therapeutic agent, where the lipid-containing particle further includes a lipidoid, cholesterol or a derivative thereof, a helper lipid, and a polyethylene glycol (PEG)- based compound. The HMO can include 3-Fucosyllactose, 6′Sialyllactose, 3′Sialyllactose (3′S), neutral penta-to-hexasaccharides, 2′Fucosyllactose, Sialylated Oligosaccharides, Lacto- N-fucopentaose I, Lacto-N-tetraose, and / or Neutral tetrasaccharides. In some cases, the HMO can be 3′S. The lipid-containing particle can be a lipid nanoparticle. The therapeutic agent can be a nucleic acid. The nucleic acid can include an RNA (e.g., one or more of a mRNA, siRNA, shRNA, miRNA, antisense RNA, guide RNA, long non-coding RNA, transfer RNA, ribosomal RNA, dsRNA, and an RNA aptamer). The RNA can be an mRNA encoding luciferase. The lipidoid can be an ionizable lipid that includes one or more of 306Oi10, 200Oi10, SM-102, DLin-MC3-DMA, ALC-0315, 306O10, 503Oi10, 402O6,10, 500X1, 500Oi10; 306O11, 306O10, 306O12, 200X6, 516i10, 500O1,1,8, 514X6, 306O14, 501X1, 205O16, 500O13, 113Oi10, 306O16, 306O13, 205O18, 509X7, 501Oi10, 503Oi10, 503O8,12, 500O14, 113Oi10, 509X1, 509X3, 501X2, 402O6,10, 516O4,8, 402X8, 501O1,1,8, or 509O1,1,8. In some cases, the lipidoid can include one or more of 306Oi10, 200Oi10, SM-102, DLin-MC3-DMA, ALC-0315, C12-200, or 503O8,12, and / or the cholesterol or derivative thereof can be cholesterol, and / or the helper lipid can be 1,2-distearoyl-sn-glycero-3-phosphocholine, and / or the PEG-based compound can be a PEG-lipid. In another aspect, this document features a method for treating a subject having an infectious disease, autoimmune disease, cardiovascular disease, respiratory disease, neurological disease, cancer, metabolic disease, musculoskeletal disease, hematologic disease, or any combination thereof. The method can include, or consist essentially of, administering to the subject an effective amount of a composition described herein. Attorney Docket No.26100-0153WO1 2023-273 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 invention pertains. Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting. The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from the description and drawings, and from the claims. DESCRIPTION OF DRAWINGS FIG.1 shows the effect of inflammation on the efficacy of LNPs for mRNA delivery. Lipopolysaccharide (LPS) was added to RAW-BLUETMcells, followed by the addition of LNPs (200Oi10with DSPC) encapsulating luciferase mRNA (mLuc) at the times indicated on the x-axis after LPS treatment. Luminescence was measured to assess efficacy using BRIGHT-GLO™ (Promega Corp.; Madison, WI). Error bars represent s.e.m. (n =5) with ** and **** representing p<0.01 and p<0.0001, respectively, according to one-way ANOVA. FIGS.2A-2C show that HMOs can reduce inflammation in cell culture. FIG.2A shows the effects of different HMOs on inflammation. The indicated HMOs were added to RAW-BLUETMcells at 10 μg / mL. LPS was added at 10 ng / mL for the positive control. Inflammation was quantified by measuring the NF-kB response. FIG.2B shows the effect of HMOs on a pre-existing inflammatory environment induced by LPS treatment. LPS was added to RAW-BLUETMcells at 10 ng / mL. After 6 hours, library containing the indicated HMOs was also added to the cells, and the NF-kB response was measured. Several HMOs statistically reduced the inflammation caused by LPS. FIG.2C shows cell viability after treatment with HMOs. LPS was added to RAW-BLUETMcells at 10 ng / mL. Six hours later, each HMO from the HMO library was also added to the cells. TritonX (TX) was added as a positive control. Cell viability was measured using an MTT assay. Error bars represent s.e.m. Attorney Docket No.26100-0153WO1 2023-273 (n =5), with ** and **** representing p<0.01 and p<0.0001, respectively, according to one- way ANOVA. FIGS.3A-3C show dose dependent effects of HMOs on inflammation. LPS at 10 ng / mL was added to RAW-BLUETMcells. Six hours later, HMOs at concentrations of 1, 10, and 100 μg / mL were added to the cells, and NF-kB was measured to quantify inflammation. 3′Sialyllactose (FIG.3A), neutral penta-to-hexasaccharides (FIG.3B), and 2′Fucosyllactose (FIG.3C) varied in their concentrations for maximal reductions in inflammation. Error bars represent s.e.m. (n = 5-15), with * and **** representing p<0.05 and p<0.0001, respectively, according to one-way ANOVA. FIGS.4A-4B show that HMOs can lead to increased LNP efficacy under inflammatory conditions. LPS at 10 ng / mL was added to RAW-BLUETMcells. HMOs and LNPs (200Oi10) were added at varying times after LPS treatment (hours after LPS treatment are indicated on the x-axis). LNP efficacy was measured by BRIGHT-GLO™ luminescence. 3′Sialyllactose was added at 1 μg / mL (FIG.4A), and 2′Fucosyllactose was added at 10 μg / mL (FIG.4B). Error bars represent s.e.m. (n = 5), with *, **, ***, and **** representing p<0.05, p<0.01, p<0.001, and p< 0.0001, respectively, according to one-way ANOVA. FIGS.5A-5I show that under inflammatory conditions, HMO treatment can increase LNP efficacy and reduce inflammation, and that HMOs are not toxic. LPS at 10 ng / mL was added to RAW-BLUETMcells, followed by the addition of LNPs 6 hours after LPS treatment. HMOs were added at varying times before or after LPS treatment (the hours after LPS treatment are indicated on the x-axis). LNPs (200Oi10) were added 6 hours after LPS treatment at 0.005 mg / mL mRNA. FIGS.5A-5C show LNP efficacy as measured by BRIGHT-GLO™ luminescence. FIGS.5D-5F show the NF-κB response, which was measured to quantify inflammation. FIGS.5G-5I show relative cell viability, which was measured using a MTT assay. 3′Sialyllactose was the HMO added at 1 μg / mL in FIGS.5A, 5D, and 5G, neutral penta-to-hexasaccharides were added at 10 μg / mL in FIGS.5B, 5E, and 5H, and 2′Fucosyllactose was added at 10 μg / mL in FIGS.5C, 5F, and 5I. Error bars represent s.e.m. (n = 5), with *, **, and **** representing p<0.05, p<0.01, and p< 0.0001, respectively, according to one-way ANOVA with Dunnett’s multiple comparison test. FIGS.6A-6D show that 3′Sialyllactose improves LNP efficacy. FIG.6A: 306Oi10LNPs encapsulating mLuc were intravenously (IV) injected into mice simultaneously along Attorney Docket No.26100-0153WO1 2023-273 with HMOs (3′Sialyllactose, neutral penta-hexasaccharides, and 2′fucosyllactose, each at 10 μg / mouse). After 6 hours, the mice were sacrificed and imaged using the IVIS® imaging system, and the total luminescent flux was measured. FIG.6B: 306Oi10 LNPs encapsulating mLuc were IV injected into mice, along with 3′Sialyllactose at a concentration from 0.5 to1000 μg / mouse. Six hours later, the mice were sacrificed and imaged using the IVIS® imaging system to determine the total luminescent flux. FIG.6C shows the organ distribution of the total luminescent flux to indicate the dose response of 3′Sialyllactose (at doses ranging from 0.5 to 1000 µg / mouse), which was injected along with 306Oi10LNPs encapsulating mLuc. FIG.6D shows the total luminescent flux that resulted from IV injection of different LNPs into mice along with 3′Sialyllactose at 10 μg / mouse. After 6 hours, the mice were sacrificed and the total luminescent flux was measured using the IVIS® imaging system. Error bars represent s.e.m. (n = 6), with *, **, ***, and **** representing p<0.05, p<0.01, p<0.001, and p< 0.0001, respectively, according to one-way ANOVA. FIGS.7A-7C show that 3′Sialyllactose rescues LNP efficacy under inflammatory conditions. FIG.7A shows the results of a kinetic study in mice. The mice received an IP injection of LPS at 0.1 mg / kg per mouse, and after 4 hours, the animals were given an IV injection of LNPs encapsulating mLuc, along with 3′Sialyllactose at 10 μg / mouse. Live images were taken using the IVIS® imaging system to measure the total luminescent flux. FIG.7B shows the total luminescent flux, indicating the response to 3′Sialyllactose at doses ranging from 0.5 to 1000 μg / mouse, administered in addition to 306Oi10 LNPs. The mice received an IP injection of LPS at a dose of 0.1 mg / kg. Four hours later, the mice received an IV injection of 306Oi10LNPs encapsulating mLuc, along with 3′Sialyllactose. After 6 hours, the mice were sacrificed and imaged using the IVIS® imaging system to determine the total luminescent flux. FIG.7C shows the total luminescent flux in the indicated organs, indicating the response to 3′Sialyllactose at the indicated doses, along with LNPs encapsulated mLuc, in mice pretreated with LPS. Error bars represent s.e.m. (n = 6), with ** and **** representing p<0.001 and p<0.0001, respectively, according to one-way ANOVA. FIGS. 8A and 8B show the effect of inflammation on LNP efficacy in vitro. FIG.8A is a graph plotting relative luminescence intensity in RAW-BLUE™ cells incubated with LPS and LNPs (306Oi10) . During 0 to 8 hours of incubation with LPS, 306Oi10LNPs loaded with firefly luciferase mRNA were applied at a dose of 100 ng / well. Luminescence was Attorney Docket No.26100-0153WO1 2023-273 measured 24 hours later. FIG.8B is a graph plotting NF-κB activity in RAW-BLUE™ cells, measured to quantify inflammation after 24 hours of incubation with LPS and LNPs. Error bars represent s.e.m. (n = 5), with *, **, and **** representing p < 0.05, p < 0.01, and p < 0.0001, respectively, according to one-way ANOVA with Tukey’s multiple comparison test. FIG.9 is a table listing the HMOs in the library used in the experiments described herein. FIGS.10A-10F show the effect of HMOs on LNP efficacy under non-inflammatory and inflammatory conditions in vitro. To probe the inherent immunogenicity of HMOs listed in the HMO library shown in FIG.9, RAW-BLUE™ cells were incubated with PBS (negative control), LPS (positive control; 10 ng / mL), or HMOs. FIG. 10A shows the NF-κB inflammatory response quantified after 24 hours of incubation with individual HMOs in the absence of LPS. To determine whether the HMOs could reduce inflammation, LPS-treated RAW-BLUE™ cells were incubated with 10 µg / mL of each individual HMO from the HMO library, and the NF-κB inflammatory response quantified after 24 hours (FIG.10B). FIG. 10C shows viability of LPS-treated RAW-BLUE™ cells, quantified after 24 hours of incubation with each HMO from the HMO library. FIGS.10D and 10E show the dose- dependent impact of 3′S (3′Sialyllactose), N5 / 6 (neutral penta-to-hexasaccharides), and 2′F (2′Fucosyllactose) on the NF-κB inflammatory response under non-inflamed (FIG. 10D) and inflamed conditions (FIG.10E). To assess if HMOs could rescue LNP efficacy under inflammation, LPS-treated RAW-BLUE™ cells were incubated with 306Oi10 LNPs at a dose of 100 ng / well, along with 3′S, N5 / 6, and 2′F at 10 μg / mL. FIG.10F shows the luminescence measured after 24 hours of incubation. Error bars represent s.e.m. (n = 5), with **, ***, and **** representing p < 0.01, p < 0.001, and p < 0.0001, respectively, according to one-way ANOVA with Dunnett’s multiple comparison test. FIGS.11A and 11B show the effect of HMOs on LNP efficacy across a time course of LPS-induced inflammatory conditions in vitro. LPS at 10 ng / mL was added to RAW- cells. HMOs and 200Oi10 LNPs were added at the indicated times after the application of LPS. LNP efficacy were measured by BRIGHT-GLO™ luminescence. FIG. 11A shows the effect of 3′Sialyllactose at a dose of 1 μg / mL. FIG.11B shows the effect of 2′Fucosyllactose at a dose of 10 μg / mL. Error bars represent s.e.m. (n = 5), with *, **, ***, Attorney Docket No.26100-0153WO1 2023-273 and **** representing p<0.05, p<0.01, p<0.001, and p<0.0001, respectively, according to one-way ANOVA. FIGS.12A-12F show the effect of 3′S on LNP efficacy under LPS-induced inflammatory conditions in vivo. FIG.12A is a schematic illustrating an experiment conducted in LPS-treated mice. First, mice received an intraperitoneal (IP) injection of LPS. Four hours later, 306Oi10LNPs encapsulating mLuc were co-delivered to the mice with 3′S via an intravenous (IV) injection at a dose of 0.5 mg / kg of mRNA and varying doses of 3′S (0-50 mg / kg). Six hours later, organs were excised and imaged for luminescence using the IVIS® imaging system. FIGS.12B and 12C include representative IVIS® images (FIG. 12B) and a graph plotting quantified total luminescent flux (FIG.12C), showing that 3′S improved LNP efficacy under pre-existing inflammation. FIG.12D shows total luminescent flux for the three most-transfected organs (n=6). FIG.12E shows the levels of cytokines and chemokines, quantified 3 hours after LNP injection using LUMINEX® Multiplex Assays (n=3). FIG. 12F includes representative images showing histological analysis for toxicity in livers and spleens harvested from mice that were euthanized 48 hours after LNP injection. Scale bars represent 100 μm. Error bars represent s.e.m., with ** and **** representing p<0.01 and p<0.0001, respectively, according to one-way ANOVA with Dunnett’s multiple comparison test (FIG. 12C), and two-way ANOVA with Tukey’s multiple comparison test (FIGS. 12D-12E). FIGS. 13A-13G show the effects of 3′S on LNP efficacy in a healthy murine model. FIG.13A is a schematic illustrating an experiment conducted in healthy mice. 306Oi10 LNPs encapsulating mLuc were co-administered with HMOs to mice via IV injection at a dose of 0.5 mg / kg of mRNA and 0-50 mg / kg of HMOs . Six hours later, organs were harvested and imaged for luminescence using the IVIS® imaging system. FIGS.13B and 13C include representative IVIS® images (FIG.13B) and a graph plotting the quantified total luminescent flux (FIG. 13C), showing that various concentrations of 3′S improved LNP baseline efficacy (n=6). FIG.13D shows Luminescent flux for the three most-transfected organs. FIG.13E shows the levels of cytokines and chemokines that were quantified using LUMINEX® Multiplex Assays (n=3) 3 hours after LNP injection. FIG.13F shows the efficacy of LNPs containing six additional ionizable lipids, with or without the HMO 3′S. FIG.13G shows total luminescent flux determined using Living Image software (n=3) when Attorney Docket No.26100-0153WO1 2023-273 306Oi10LNPs were co-administered with 3′S, 5 / 6, or 2′F at a dose of 0.5 mg / kg. Error bars represent s.e.m., with *, **, ***, and **** representing p<0.05, p<0.01, p<0.001, and p<0.0001, respectively, according to one-way ANOVA with Dunnett’s multiple comparison test (FIGS.13C and 13G), and two-way ANOVA with Tukey’s multiple comparison test (FIGS. 13D-13F). FIG.14 shows the effects on LNP efficacy in vitro when incorporating HMOs into LNP formulation. HMOs were incorporated into the LNP either in an aqueous solution, an mRNA solution, or a lipid solution during the formulation of the LNPs. 200Oi10LNPs with either DOPE or DSPC were loaded with firefly luciferase mRNA and were applied at a dose of 100 ng / well, where the lipidoid:mRNA (weight / weight) ratio was 20:1. Luminescence was measured 24 hours later. The relative luminescence intensity in RAW-BLUE™ cells incubated LNPs was plotted against various formulation of 200Oi10 LNPs with either DOPE or DSPC. Error bars represent s.e.m. (n = 5), with *, **, *** and **** representing p < 0.05, p < 0.01, p < 0.001, and p < 0.0001, respectively, according to one-way ANOVA with Dunnett’s multiple comparison test. DETAILED DESCRIPTION COMPOSITIONS Provided herein are compositions containing one or more human milk oligosaccharides (HMOs) and a lipid-containing particle (e.g., LNP) for delivery of one or more therapeutic agents (e.g., a nucleic acid, such as an mRNA). As used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. As used herein, the term “about,” when used herein in reference to a value, refers to a value that is ± 10% of the referenced value. As used herein, “nucleic acid” is used to include any compound and / or substance that comprises a polymer of nucleotides. In some embodiments, polymers of nucleotides are referred to as polynucleotides. Exemplary nucleic acids or polynucleotides include, but are not limited to, ribonucleic acids (RNAs), deoxyribonucleic acids (DNAs), threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), locked nucleic Attorney Docket No.26100-0153WO1 2023-273 acids (LNAs, including LNA having a β-D-ribo configuration, α-LNA having an a-L-ribo configuration (a diastereomer of LNA), 2’-amino-LNA having a 2’-amino functionalization, and 2’-amino-a-LNA having a 2’-amino functionalization) or hybrids thereof. Naturally occurring nucleic acids generally have a deoxyribose sugar (e.g., found in deoxyribonucleic acid (DNA)) or a ribose sugar (e.g., found in ribonucleic acid (RNA)). Nucleic acids and nucleic acid analogs comprise a backbone and a sequence of nucleobases. In the context of the present disclosure, the backbone monomer residues can be any suitable nucleic acid backbone monomer residues having a negative charge, such as a ribose or deoxyribose connected to another ribose or deoxyribose by a phosphodiester bond, or a backbone residue of a nucleic acid analog monomer. The backbone monomer includes both the structural “residue” component, such as the ribose in RNA, and any active groups that are modified in linking monomers together, such as the 5’ triphosphate and 3’ hydroxyl groups of a ribonucleotide, which are modified when polymerized into RNA to leave a negatively-charged phosphodiester linkage. As used herein, a “therapeutic agent” is any compound or composition that can be delivered to a patient to achieve a desired effect, such as a beneficial, treatment, or curative effect. Therapeutic agents include, without limitation, nucleic acids, nucleic acid analogs, proteins, polypeptides, small molecule drugs, antibiotics, antivirals, and cell-based therapies (e.g., CAR-T cell therapies). For example, therapeutic agents can be nucleic acids or nucleic acid analogs having a negatively-charged backbone, including but not limited to single- stranded DNA, single-stranded RNA, double-stranded DNA, double-stranded RNA, or modified versions of any of the preceding (e.g., versions that include one or more changes to the nucleotide components), or analogs of any of the preceding (e.g., synthetic molecules that mimic the structure and function of the original). With regard to overall structure and function of the nucleic acid or nucleic acid analog, the nucleic acid or nucleic acid analog may be, without limitation: mRNA (messenger RNA), siRNA (small interfering RNA), miRNA (microRNA), gRNA (guide RNA), tRNA (transfer RNA), rRNA (ribosomal RNA), tmRNA (transfer-messenger RNA), lncRNA (long non-coding RNA), circRNA (circular RNA), antisense RNA, ncRNA (non-coding RNA), telomerase RNA, piRNA (Piwi- interacting RNA), snRNA (small nuclear RNA), snoRNA (small nucleolar RNA), scaRNAs (small Cajal body RNA), Y RNA, eRNA (enhancer RNA), shRNA (small hairpin RNA), Attorney Docket No.26100-0153WO1 2023-273 stRNA (small temporal RNA), DNA, chloroplast DNA, cDNA (complementary DNA), gDNA (genomic DNA), Hachimoji DNA, mitochondrial DNA, msDNA (multicopy single- stranded DNA), XNA (xeno nucleic acid), glycol nucleic acid, threose nucleic acid, hexose nucleic acid, LNA (locked nucleic acid), PNA (peptide nucleic acid), morpholino oligomer, antisense oligonucleotide, ribozyme, deoxyribozyme, aptamer, cloning vector, phagemid, plasmid, lambda phage, cosmid, fosmid, or artificial chromosome. RNA Therapeutics RNA therapeutics are a class of RNA-based treatments that target specific genes or genetic pathways with high specificity. The use of RNA therapeutics allow for transient expression or inhibition, which can reduce long-term side effects. RNA therapeutics utilize various forms of RNA to treat diseases such as, without limitation, infectious diseases, cancer, genetic disorders, cardiovascular diseases, and neurological diseases. Research has been ongoing since the 1990s, with significant success in cancer therapy in the early 2010s (see, e.g., Sahin et al., Nature Reviews Drug Discovery, 13:759-780, 2014). The RNA used in an RNA therapeutic can include, for example, mRNA, siRNA, short hairpin RNA (shRNA), microRNA (miRNA), antisense RNA, gRNA, long non-coding RNA, transfer RNA, ribosomal RNA, double-stranded RNA (dsRNA), and / or an RNA aptamer. In some cases, for example, an mRNA-based therapy can be used. mRNA-based therapies can trigger synthesis of proteins by delivering coding mRNA into cells, making such therapies particularly useful in vaccine development (see, e.g., DeFrancesco, Nature Biotechnology, 35:193-197, 2017). The coding mRNA can be designed as a blueprint to generate a protein of interest (e.g., a reporter protein, a functional protein, or an antigen). In some embodiments, a protein of interest can be an antigen produced by a pathogen (e.g., a virus) or by a cancer cell. Such protein molecules can stimulate an adaptive immune response that teaches the body to identify and destroy the corresponding pathogen or cancer cells (see, e.g., Bae and Park, Advanced Drug Delivery Reviews, 158:4-16, 2020). mRNA vaccines (e.g., the Pfizer–BioNTech COVID-19 vaccine and the Moderna COVID-19 vaccine) were developed for use in combating the coronavirus disease during the COVID-19 pandemic (see, Noor, Current Clinical Microbiology Reports, 8(3):178-185, 2021). As described herein, the designed mRNA encodes firefly luciferase as a Attorney Docket No.26100-0153WO1 2023-273 reporter protein. An mRNA encoding a desired protein (e.g., an mRNA encoding luciferase) can be delivered into cells using lipid-containing particles (e.g., LNPs) to produce the protein (e.g., luciferase) in vitro, through cell culture, and in vivo, such as in mouse models or in any other appropriate mammal (e.g., humans, non-human primates, rats, rabbits, cows, pigs, sheep, dogs, and / or cats). The mammal can be healthy or can have, for example, an inflammatory condition. Challenges in the application of mRNA-based therapeutics can include efficiency of mRNA delivery, mRNA stability, and immune response of a recipient against the therapeutics. Successful mRNA-based therapeutics typically rely on efficiently delivering mRNA molecules into target cells while avoiding degradation of the mRNAs by nucleases, enhancing mRNA stability to ensure they reach and remain functional at their targets, and minimizing the immune responses to exogenous mRNA, which can reduce or prevent inflammation and / or other adverse effects. LNPs and components thereof Lipid-containing particles are small particles or structures that include lipids and / or lipid-like materials (e.g., lipidoids). Lipids include, without limitation, fats, waxes, sterols, fat-soluble vitamins, and other similar substances. Lipidoids are a class of lipid-like materials often used in biotechnology and nanomedicine, particularly for the delivery of nucleic acids, such as RNA and DNA. Lipid-containing particles can be found in various biological forms such as lipid nanoparticles (LNPs, used in drug delivery systems), lipoproteins (which transport lipids in the bloodstream), lipid droplets (intracellular storage organelles), exosome (involved in cell-to-cell communication), vesicles (small membrane-bound sacs within cells), and others. Lipid-containing particles (e.g., LNPs) have been shown to be effective for delivering nucleic acids and nucleic acid analogs to cells, often in vivo (see, e.g., Kulkarni et al., Nucleic Acid Therapeutics, 28(3):146-157, 2018; Hajj and Whitehead, Nature Reviews Materials, 2:17056, 2017; U.S. Publication No.20130245107; and U.S. Patent No.8,754,062). In various embodiments, a lipid-containing particle (e.g., a LNP) can include a mixture of an ionizable and / or cationic lipid or lipidoid, cholesterol or a cholesterol analogue, a helper lipid such as DSPC, DOPC (dioleoyl phosphatidylcholine), DSPE (distearoyl Attorney Docket No.26100-0153WO1 2023-273 phosphatidylethanolamine), or DOPE, and a polyethylene glycol-lipid conjugate (PEG-lipid) or PEG-cholesterol conjugate (PEG-cholesterol), as well as a polyanionic therapeutic agent such as a nucleic acid or nucleic acid analog. For targeting purposes, the PEG-lipid or PEG- cholesterol can be modified with a targeting moiety, such as N-acetylgalactosamine (GalNAc) for liver targeting, or with another ligand or binding reagent, such as an antibody or antibody fragment. The lipid-containing particles (e.g., LNPs) in the compositions provided herein can include any appropriate lipid or lipidoid, cholesterol or cholesterol derivative, helper lipid, and polyethylene glycol (PEG)-based compound. Lipidoids are ionizable lipids, and non- limiting examples of suitable lipidoids include 306Oi10, 200Oi10, SM-102, DLin-MC3-DMA, ALC-0315, 306O10, 503Oi10, 402O6,10, 500X1, 500Oi10; 306O11, 306O10, 306O12, 200X6, 516i10, 500O1,1,8, 514X6, 306O14, 501X1, 205O16, 500O13, 503O8,12, 113Oi10, 306O16, 306O13, 205O18, 509X7, 501Oi10, 503Oi10, 500O14, 113Oi10, 509X1, 509X3, 501X2, 402O6,10, 516O4,8, 402X8, 501O1,1,8, 509O1,1,8, and any combination thereof. In some embodiments, the lipidoid(s) in a LNP can include 306Oi10, 200Oi10, SM-102, DLin-MC3-DMA, ALC-0315, or any combination thereof. For example, the lipidoid contained in a LNP can be 306Oi10, 200Oi10, SM-102, DLin-MC3-DMA, or ALC-0315. Helper lipids are cationic, anionic, neutral, or zwitterionic amphiphilic lipids that, along with cholesterol or a derivative thereof (e.g., a cholesterol analog), can aid in the molecular packing and stability of a lipid-containing particle (e.g., a LNP). Helper lipids also can enhance lipid nanoparticle efficacy by promoting fusion with both cell and endosomal membranes, facilitating cell uptake and endosomal release. Non-limiting examples of suitable helper lipids include DSPC, DSPE, DOPC, and DOPE. Additional useful helper lipids include phosphatidylcholine lipids. For example, a helper lipid can include 1,2- distearoyl-sn-glycero-3-phosphocholine. Non-limiting examples of suitable cholesterol derivatives (or analogues) include oxidized cholesterol, desmosterol, 7-dehydrocholesterol, ergosterol, lanosterol, ketosterone, cholesterol sulfate, dehydroergosterol, cholestratrienol, 5- cholestene, and pregnenolone. In some cases, the lipid-containing particles (e.g., LNPs) in the compositions provided herein can contain cholesterol. PEG is a polyether compound derived from petroleum. PEG and PEG-based compounds can be used for various applications, such as drug delivery agents, solvents, Attorney Docket No.26100-0153WO1 2023-273 adhesives, adsorbents, and tissue engineering scaffolds. PEG-based compounds include, without limitation, PEG-lipids (PEGylated lipids) and PEG-cholesterols (PEGylated cholesterols). PEG-lipids include a PEG moiety attached to one or more lipid moieties (e.g., a ceramide, succinoyl, or carbamate moiety). PEG-cholesterols include a PEG moiety attached to one or more cholesterol moieties. PEG-lipids and / or PEG-cholesterols can form a protective, non-aggregating, non-immunogenic shell around the surface of LNPs. Depending on the ultimate delivery route of the LNPs, the lipid group may be varied (e.g., in length) to dictate how long the PEG-lipid will be associated with the LNP, with longer lipid chains tending to remain associated with the LNP for longer time periods, and shorter lipid chains typically being useful for providing “diffusible” PEG lipids that diffuse from the lipid nanoparticle quickly to produce an LNP with increased transfection rates. The PEG moiety of a PEG-lipid or PEG-cholesterol can have a molecular weight ranging from about 300 g / mol to about 5000 g / mol (e.g., about 2000 g / mol, which is referred to as PEG 2000). Non-limiting examples of suitable PEG-lipids include 1 ,2-dimyristoyl-sn-glycero-3- phosphoethanolamine-N [methoxy(polyethylene glycol)-2000], N-octanoyl-sphingosine-1- {succinyl[methoxy(polyethylene glycol)2000]}, N-palmitoyl-sphingosine-1- {succinyl[methoxy(polyethylene glycol)5000]}, 1,2- dipalmitoyl-sn-glycero-3- phosphoethanolamine-N-[methoxy(polyethylene glycol)-3000] (ammonium salt), 1,2- dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-1000] (ammonium salt), and PEG-cholesterol, such as cholesterol-(polyethylene glycol-600). In some cases, the lipid-containing particles (e.g., LNPs) in the compositions provided herein can contain a PEG-lipid. Human milk oligosaccharides (HMOs) HMOs are complex carbohydrates found in human breast milk. HMOs are composed of five different monosaccharides: glucose (Glc), galactose (Gal), N-acetylglucosamine (GlcNAc), fucose (Fuc), and N-acetylneuraminic acid (Neu5Ac, which is the prevalent form of sialic acid in humans). Lactose at the reducing end serves as the backbone of all HMOs. The structural diversity of HMOs is due to combinations of elongation with N- acetyllactosamine, Lacto-N-biose, fucose and / or sialic acid. Lactose can be extended through the addition of disaccharides lacto-N-biose (Galβ1,3GlcNAc) or N-acetyllactosamine Attorney Docket No.26100-0153WO1 2023-273 (Galβ1,4GlcNAc). While the addition of lacto-N-biose stops the growth of the chain, addition of N-acetyllactosamine allows further elongation. The branching of HMOs occurs with the addition of a β1,6 linkage between two disaccharides. The addition of fucose (fucosylation) onto lactose via α1,2 or α1,3 linkage results in trisaccharide 2′FL or 3FL, while lactose elongation by the addition of Neu5Ac (sialylation) via an α2,3 or α2,6 linkage produces the trisaccharides 3′SL and 6′SL. (See, Chutipongtanate, Biomedicines, 10, 2022). As described herein, examples of HMOs that can be included in a composition provided herein and / or used in the methods provided herein include, but are not limited to, 3- Fucosyllactose (3′F), 6′Sialyllactose (6′S), 3′Sialyllactose (3′S), neutral penta-to- hexasaccharides (N5 / 6), 2′Fucosyllactose (2′F), Sialylated Oligosaccharides, Lacto-N- fucopentaose I, Lacto-N-tetraose, and Neutral tetrasaccharides. For example, the compositions provided herein can contain 3′S, 2′F, N5 / 6, or any combination thereof, and the methods provided herein can include delivering to a subject (e.g., mammal) 3′S, 2′F, N5 / 6, or any combination thereof. Methods for making LNP-HMO compositions Any appropriate method can be used to prepare a composition that contains a lipid- containing particle (e.g., an LNP) and one or more HMOs. For example, an LNP preparation can be generated by combining an ionizable lipid or lipidoid, cholesterol or a cholesterol derivative, a helper lipid, and a PEG-based compound in any appropriate amounts or ratios to generate LNPs. In some cases, LNPs can be prepared by combining: about 30 mol% to about 70 mol% (e.g., about 30 to 40 mol%, about 35 to 45 mol%, about 40 to 50 mol%, about 45 to 55 mol%, about 50 to 60 mol%, about 55 to 65 mol%, about to 60 to 70 mol%, about 30 mol%, about 40 mol%, about 50 mol%, about 60 mol%, or about 70 mol%) of a lipid or lipidoid (e.g., 306Oi10); about 2 mol% to about 25 mol% (e.g., about 2 to 10 mol%, about 5 to 15 mol%, about 10 to 20 mol%, about 5 mol%, about 10 mol%, or about 15 mol%) of a helper lipid (e.g., DSPC); about 20 to about 60 mol% (e.g., about 20 to 30 mol%, about 25 to 35 mol%, about 30 to 40 mol%, about 35 to 45 mol%, about 40 to 50 mol%, about 45 to 55 mol%, about 50 Attorney Docket No.26100-0153WO1 2023-273 to 60 mol%, about 30 mol%, about 35 mol%, about 38.5 mol%, about 40 mol%, about 45 mol%, or about 50 mol%) cholesterol or a cholesterol derivative (e.g., cholesterol); and about 0.2 to about 3 mol% (e.g., about 0.2 to 1 mol%, about 0.5 to 1.5 mol%, about 1 to 2 mol%, about 1.5 to 2.5 mol%, about 2 to 3 mol%, about 0.5 mol%, about 1 mol%, about 1.5 mol%, about 2 mol% or about 2.5 mol%) of a PEG-based compound (e.g., PEG). LNPs prepared as described herein can be combined with any appropriate cargo (e.g., a nucleic acid, as described herein) and, in some cases, with one or more HMOs (e.g., 3′S, N5 / 6, 2′F, any of the other HMOs disclosed herein, or any combination thereof). One or more HMOs can be added to an LNP preparation at any appropriate amount (e.g., about 0.5 μg / mL, about 1 μg / mL, about 2 μg / mL, about 5 μg / mL, about 10 μg / mL, about 15 μg / mL, about 20 μg / mL, about 50 μg / mL, about 100 μg / mL, about 200 μg / mL, about 500 μg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, or more than about 5 mg / mL). The HMO can be added with the cargo, lipid solutions, aqueous LNP solutions, and / or separate from the LNP. In some cases, the concentration of mRNA as a cargo in an LNP (e.g., for in vitro cell culture) can be from about 0.001 mg / mL to about 2 mg / mL (e.g., from about 0.001 mg / mL to about 1.5 mg / mL, from about 0.003 mg / mL to about 1.5 mg / mL, from about 0.005 mg / mL to about 1.5 mg / mL, from about 0.003 mg / mL to about 1 mg / mL, from about 0.003 mg / mL to about 0.5 mg / mL, from about 0.005 to about 1.5 mg / mL, from about 0.005 to about 1 mg / mL, from about 0.005 to about 0.5 mg / mL, about 0.001 mg / mL, about 0.003 mg / mL, about 0.005 mg / mL, about 0.01 mg / mL, about 0.03 mg / mL, about 0.05 mg / mL, about 0.08 mg / mL, about 0.1 mg / mL, about 0.2 mg / mL, about 0.3 mg / mL, about 0.4 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 1.5 mg / mL, or about 2 mg / mL). In some cases, the concentration of mRNA as a cargo in an LNP (e.g., for in vivo use) can be from about 0.01 mg / mL to about 10 mg / mL (e.g., from about 0.01 mg / mL to about 1 mg / mL, from about 0.03 mg / mL to about 3 mg / mL, from about 0.05 mg / mL to about 5 mg / mL, from about 0.1 mg / mL to about 2 mg / mL, from about 0.3 mg / mL to about 3 mg / mL, from about 0.5 mg / mL to about 5 mg / mL, from about 1 mg / mL to about 3 mg / mL, from about 3 to about 5 mg / mL, about 0.01 mg / mL, about 0.03 mg / mL, about 0.05 mg / mL, about 0.1 mg / mL, about 0.3 mg / mL, about 0.5 mg / mL, about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 Attorney Docket No.26100-0153WO1 2023-273 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, or about 10 mg / mL). As described herein, the ratio of lipidoid:mRNA (weight / weight) can be about 5 to about 50 (e.g., about 5 to about 40, 5 to about 30, about 5 to about 20 about, 10 to about 50, about 10 to about 40, about 10 to about 30, about 10 to about 20, about 5, about 10, about 12.5, about 15, about 17.5, about 20, about 25, about 30, about 40, or about 50). Assessing effectiveness of LNP-HMO combinations Any appropriate method can be used to determine the effectiveness of a lipid- containing particle (e.g., LNP) and a HMO (e.g., a LNP and a HMO delivered together in the same composition, or a LNP and a HMO delivered separately). In some cases, the effectiveness of a composition can be assessed in the presence of inflammation (e.g., inflammation induced by lipopolysaccharide (LPS). LPS is a component of the outer membrane of Gram-negative bacteria, and is a potent inducer of inflammation. LPS-induced inflammation can be used in experimental in vitro and in vivo models to study the mechanisms of inflammation and to test anti-inflammatory drugs. In in vitro models, for example, cultured immune cells (e.g., macrophages and / or dendritic cells) can be treated with LPS to study cytokine production and signaling pathways. In in vivo models, animal models (e.g., mice) can be injected with LPS to induce systemic inflammation or inflammation of one or more specific organs (e.g., lung, liver, and / or spleen). One or more test compositions then can be administered and their effects can be assessed. Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a protein complex that acts as a transcription factor, playing a crucial role in regulating inflammatory responses. In acute inflammation, NF-κB levels rapidly increase in response to inflammatory stimuli. This leads to a swift production of pro-inflammatory mediators that help to eliminate pathogens and initiate tissue repair. Persistent activation of NF-κB is a hallmark of chronic inflammatory diseases. Elevated NF-κB levels and prolonged activity can result in continuous production of pro-inflammatory mediators, contributing to tissue damage and disease progression. NF-κB levels can be measured to determine the effect of a composition (or a combination of compositions, such as a LNP and a HMO delivered separately) on inflammation, such that a drop in NF-κB levels after administration of one or more test Attorney Docket No.26100-0153WO1 2023-273 compositions can indicate that the composition(s) are effective to reduce inflammation. Any appropriate method can be used to determine a level of NF-κB in cells or tissues of a mammal, for example. Suitable techniques for measuring NF-κB levels include, without limitation, western blotting to measure the levels of NF-κB subunits (e.g., p65 and / or p50) and their phosphorylated forms in cell or tissue extracts, electrophoretic mobility shift assay, ELISA (enzyme-linked immunosorbent assay) to measure the levels of NF-κB subunits or phosphorylated IκB in cell lysates, immunofluorescence and immunohistochemistry, and quantitative PCR to measure expression levels of NF-κB target genes as an indirect indicator of NF-κB activity. As described herein, RAW-BLUETMcells can be used as cultured immune cells in studies to assess the effectiveness of a composition or a combination of compositions on LPS-induced inflammation. For example, RAW-BLUETMcells, derived from murine macrophages, can be used to monitor the NF-κB and AP-1 (Activator Protein-1) responses to LPS stimulation. As demonstrated herein, NF-κB can be measured by QUANTI-BlUE™ Solution assay (InvivoGen) to indicate the level of LPS induced inflammation in RAW- BLUETMcells. For example, RAW-BLUETMcells can be modified to express secreted embryonic alkaline phosphatase (SEAP) which is a downstream target of the transcription factor NF-κB. NF-κB activation induced by inflammatory agents prompts SEAP secretion into the cell culture supernatant, which can be detected using the QUANTI-BLUE™ assay. The detection value corresponds to the level of NF-κB activity, and thus can indicate inflammatory activity. METHODS This document also provides methods for delivering a therapeutic agent (e.g., an mRNA). The methods can include administering to a subject an HMO and a lipid-containing particle (e.g., LNP) that encapsulates the therapeutic agent (e.g., mRNA). The HMO and the LNP encapsulating the therapeutic agent can be delivered together, in a single composition, or can be administered separately. In some cases, one or more HMOs can be administered prior to an LNP encapsulating a therapeutic agent. In some cases, an LNP encapsulating a therapeutic agent can be administered prior to one or more HMOs. When an LNP is administered separately from an HMO, their delivery can be separated by any appropriate Attorney Docket No.26100-0153WO1 2023-273 length of time (e.g., about 30 to 60 minutes, about 1 to 2 hours, about 2 to 4 hours, about 4 to 8 hours, about 8 to 12 hours, about 12 to 24 hours, about 1 to 2 days, about 2 to 3 days, about 3 to 5 days, or about 5 to 7 days). Methods of RNA delivery Methods that include delivering RNA into cells can be useful in research and therapeutic applications, including gene silencing, gene editing, and mRNA-based therapeutics. As described herein, RNA delivery can be achieved via LNPs, which can avoid issues encountered with delivery of naked, single-stranded RNA (which is prone to nuclease degradation, can activate the immune system, and is too large and negatively charged to passively cross the cell membrane). The methods provided herein can be used for delivering a therapeutic agent to a subject (e.g., a mammal such as a human). The methods can include administering to the subject an HMO and a lipid-containing particle (e.g., a LNP) containing the therapeutic agent. The HMO can include one or more of 3-Fucosyllactose, 6′Sialyllactose, 3′Sialyllactose (3′S), neutral penta-to-hexasaccharides, 2′Fucosyllactose, Sialylated Oligosaccharides, Lacto- N-fucopentaose I, Lacto-N-tetraose, and Neutral tetrasaccharides. In some cases, 3′Sialyllactose (3′S), neutral penta-to-hexasaccharides (N5 / 6), and / or 2′Fucosyllactose (2′F) can be used to reduce inflammation. As demonstrated herein, for example, 3′S can improve the efficacy of LNP-mediated mRNA delivery under both healthy and inflammatory conditions. An HMO (e.g., 3′S) or a composition containing more than one HMO can be administered via any appropriate route. For example, one or more HMOs can be administered to a subject (e.g., a mammal) intravenously, intramuscularly, or subcutaneously. Further, an HMO can be administered in any appropriate amount. In some cases, an HMO (e.g., 3′S) can be administered at a dose of about 0.01 to about 100 mg / kg (e.g., about 0.01 to about 50 mg / kg, about 0.05 mg / kg to about 25 mg / kg, about 0.1 mg / kg to about 20 mg / kg, about 0.5 mg / kg to about 15 mg / kg, about 1 mg / kg to about 10 mg / kg, about 0.025 mg / kg, about 0.1 mg / kg, about 0.25 mg / kg, about 0.5 mg / kg, about 0.75 mg / kg, about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, Attorney Docket No.26100-0153WO1 2023-273 or about 100 mg / kg. For example, the HMO 3′S can, in some embodiments, be administered at a dose of about 0.5 mg / kg. A lipid-containing particle (e.g., a LNP) can be administered to a subject (e.g., a mammal) any appropriate route. For example, a lipid-containing particle (e.g., a LNP) can be administered intravenously, intramuscularly, or subcutaneously. A lipid-containing particle (e.g., a LNP) can be administered to a subject at a mRNA dose of about 0.05 to about 5 mg / kg. For example, a lipid-containing particle (e.g., a LNP) can be administered at a mRNA dose of about 0.05 to about 5 mg / kg (e.g., about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1, about 2, about 3, about 4, or about 5 mg / kg). In some embodiments, a lipid-containing particle (e.g., a LNP) can be administered at a mRNA dose of about 0.5 mg / kg. In some cases, a HMO (e.g., 3′S) and a lipid-containing particle (e.g., LNP) can be administered simultaneously (e.g., in a composition that contains both the LNP and the HMO, or as separate compositions). In some cases, a HMO (e.g., 3′S) and a lipid-containing particle (e.g., LNP) can be administered sequentially. For example, a HMO (e.g., 3′S) can be administered about 30 minutes to about 1 week before administration of the lipid-containing particle (e.g., LNP). For example, a HMO (e.g., 3′S) can be administered about 30 to 60 minutes, about 1 to 2 hours, about 2 to 4 hours, about 4 to 8 hours, about 8 to 12 hours, about 12 to 24 hours, about 1 to 2 days, about 2 to 3 days, about 3 to 5 days, or about 5 to 7 days before administration of the lipid-containing particle (e.g., LNP). In some cases, a HMO (e.g., 3′S) can be administered about 30 minutes to about 1 week after administration of the lipid-containing particle (e.g., LNP). For example, a HMO (e.g., 3′S) can be administered about 30 to 60 minutes, about 1 to 2 hours, about 2 to 4 hours, about 4 to 8 hours, about 8 to 12 hours, about 12 to 24 hours, about 1 to 2 days, about 2 to 3 days, about 3 to 5 days, or about 5 to 7 days after administration of the lipid-containing particle (e.g., LNP). As described herein, the therapeutic agent contained within the lipid-containing particle (e.g., LNP) used in the methods provided herein can be a nucleic acid (e.g., an RNA). The RNA as the therapeutic agent can be a mRNA, siRNA, shRNA, miRNA, antisense RNA, guide RNA, long non-coding RNA, transfer RNA, ribosomal RNA, dsRNA, or RNA aptamer. In some cases, an RNA encoding a marker (e.g., an mRNA encoding a luciferase polypeptide) can be used in the methods described herein. Luciferase is an enzyme Attorney Docket No.26100-0153WO1 2023-273 that catalyzes a bioluminescent reaction, producing light as a byproduct. This reaction can be utilized in various biological and medical research applications, particularly in reporter assays to study gene expression and cellular processes. The bioluminescence produced by luciferase encoded by the mRNA encapsulated in the lipid-containing particle (e.g., LNP) can be used to assess efficacy of mRNA delivery. Methods of treating In some cases, the methods described herein can be used to treat a clinical disorder in a subject (e.g., mammal). Disorders that can be treated according to methods provided herein include, without limitation, disorders associated with inflammatory conditions, and diseases in which one or more genes or proteins is dysregulated. In some embodiments, a disorder that can be treated as described herein can be associated with inflammation in a lung, liver, spleen, blood vessel, brain, lymph node, or gastrointestinal tract of a subject (e.g., a mammal). The inflammatory condition can be induced by a stimulator such as LPS or lipid- containing particles (e.g., LNPs), or can be due to an infectious disease, injury, toxin, chronic disease (e.g., cancer), smoking, or any combination thereof. In some embodiments, the inflammation stimulator is LPS or lipid-containing particles. In some embodiments, the disorder is an infectious disease, an autoimmune disease, a cardiovascular disease, a respiratory disease, a neurodegenerative disease, a cancer, a genetic disorder a metabolic disease, a musculoskeletal disease, a hematologic disease, or any combination thereof. Generally, the methods provided herein include administering therapeutically effective amounts of one or more HMOs (e.g., 3′S) and a lipid-containing particle (e.g., LNP) as described herein, to a subject that is in need thereof or has been determined to be in need of, such treatment. The lipid-containing particle (e.g., LNP) encapsulates a therapeutic agent for the treatment needed. The therapeutic agent can be mRNA, siRNA, shRNA, miRNA, antisense RNA, guide RNA, long non-coding RNA, transfer RNA, ribosomal RNA, dsRNA, or RNA aptamers. Effective doses can vary depending on the severity of the disorder, the route of administration, the age and general health condition of the subject, excipient usage, the possibility of co-usage with other therapeutic treatments, and the judgment of the treating clinician. An effective amount of a composition containing one or more HMOs and / or a Attorney Docket No.26100-0153WO1 2023-273 lipid-containing particle (e.g., LNP) as described herein can be any amount that reduces one or more symptoms of the disorder (e.g., by at least 10, 25, 35, 45, 50, 55, 65, 75, 80, 90, or 100 percent) within a subject (e.g., a mammal), without producing severe toxicity in the mammal. As described herein, for example, an effective dose of a HMO can be from about 0.01 mg / kg to about 100 mg / kg, and an effective dose of a lipid-containing particle (e.g., LNP) can be an mRNA dose of about 0.05 to about 5 mg / kg. The effective amounts can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amounts used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple treatment agents, route of administration, severity of disorder, or risk level for development of the same or another disorder in the mammal being treated may require an increase or decrease in the actual effective amount administered. If a particular mammal fails to respond to a particular amount of a HMO and / or a lipid-containing particle (e.g., LNP), then the amount of the HMO and / or the lipid-containing particle can be increased by, for example, two-fold. After receiving the higher amount, the mammal can be monitored for both responsiveness to the treatment and toxicity symptoms, and adjustments can be made accordingly. The effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal’s response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the frequency of administration, duration of treatment, use of multiple HMOs, route of administration, and severity of the disorder may require an increase or decrease in the actual effective amount administered. The frequency of administration of one or more HMOs and one or more lipid- containing particles (e.g., LNPs) to a subject (e.g., a mammal) can be any frequency that reduces a symptom of a disorder in the subject, without producing significant toxicity to the subject. For example, the frequency of administration of a HMO and a lipid-containing particle (e.g., LNP), either individually or together, can be from about four times daily to about once a day, from about once daily to three times a week, from about three times a week to about twice a week, from about twice a week to about once a week, or from about once a week to about once a month (e.g., from about once a week to about once every other week). in some cases, a HMO (or a composition containing a HMO) can be administered at a first Attorney Docket No.26100-0153WO1 2023-273 frequency (e.g., twice daily) and a LNP (or a composition containing LNP) can be administered at a second frequency (e.g., once daily). The frequency of administration can remain constant or can be variable during the duration of treatment. A course of treatment with one or more HMOs and a lipid-containing particle (e.g., LNP) described herein can include rest periods. For example, a HMO and a LNP can be administered daily over a one- week period followed by a one-week rest period, and such a regimen can be repeated multiple times. As with the effective amount, various factors can influence the actual frequency of administration used for a particular application. For example, the effective amount, duration of treatment, use of multiple HMOs, route of administration, and severity of the disorder may require an increase or decrease in administration frequency. An effective duration for administering one or more HMOs and one or more lipid- containing particles (e.g., LNPs) to a subject (e.g., a mammal) can be any duration that reduces a symptom of a disorder in the mammal, without producing significant toxicity to the mammal. In some cases, the effective duration can vary from several days to several months. Multiple factors can influence the actual effective duration used for a particular treatment. For example, an effective duration can vary with the effective amount, frequency of administration, use of multiple HMOs, route of administration, and severity of the disorder being treated. In some cases, the progression of a disorder in a subject (e.g., a mammal) or the severity of one or more symptoms related to the disorder in the subject being treated can be monitored. Any appropriate method can be used to determine whether or not a subject having a disorder is effectively being treated. As used in this context, to “treat” means to ameliorate at least one symptom of the disorder associated with inflammatory conditions. For example, a treatment can result in a reduction in NF-κB levels. Administration of a therapeutically effective amount of a HMO as described herein can result in rescuing and / or improving LNP efficacy of mRNA delivery. Thus, the methods described herein provide an approach to enhance the effectiveness of RNA therapeutics. The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims. Attorney Docket No.26100-0153WO1 2023-273 EXAMPLES Example 1 Identification of human milk oligosaccharides (HMOs) that can reduce inflammation in murine cell lines. LNPs can have reduced delivery efficacy under inflammatory conditions, possibly because inflammation can hinder the protein translational process (FIG. 1). Studies were conducted to determine whether human milk oligosaccharides (sugars), which can have anti- inflammatory properties, would recover or improve the efficacy of RNA delivery by LNPs and subsequent protein translation. First, experiments were conducted to determine whether milk sugars could reduce inflammation in the absence of LNPs. For these in vitro screens, nine oligosaccharides from human milk were tested. These sugars were added to RAW- BLUETMcells and NF-κB activity was measured as a proxy for immune activation, indicating that HMOs can stimulate the immune system themselves (FIG. 2A). Next, the ability of HMOs to inhibit NF-κB activity in a pre-existing inflammatory environment was tested using LPS-stimulated cells (FIG.2B). Finally, the toxicity of these sugars on these cells was assessed (FIG.2C). From this screen, it was established that the most promising sugars for reduction of inflammation were 3′Sialyllactose, neutral penta-to-hexasaccharides, and 2′fucosyllactose. To determine if these sugars had dose responsive effects under an inflammatory state, the concentrations of these sugars were varied from 1 to100 μg / mL for LPS-stimulated cells and non-inflamed cells, and NF-κB activity was measured. The results of these studies showed that the most effective dose of 3′Sialyllactose was 1-10 μg / mL (FIG. 3A), while the most effective dose of neutral penta-to-hexasaccharides and 2′fucosyllactose was 10 μg / mL (FIGS.3B and 3C). Combining human milk oligosaccharides with LNPs to rescue mRNA delivery efficacy under inflammatory conditions in murine cell lines. LNPs are made up of four components: ionizable lipid, 1,2-distearoyl-sn-glycero-3- phosphocholine (DSPC), cholesterol, and polyethylene glycol (PEG)-lipid. The ionizable lipid 200Oi10, which is representative of common lipids used for LNP delivery, was used for in vitro studies at 50 mol%. DSPC at 10 mol%, cholesterol at 38.5 mol%, and PEG at 1.5 mol% also were used. The LNPs were utilized to deliver mRNA encoding luciferase in Attorney Docket No.26100-0153WO1 2023-273 studies to evaluate the timing and kinetics of treating cells with 10 ng / mL LPS along with the HMOs 3′Sialyllactose (FIG.4A) and 2′fucosyllactose (FIG. 4B). The results demonstrated that adding LNPs 6 hours after LPS treatment reduced LNP efficacy and that the addition of HMOs rescued and / or improved LNP efficacy under inflammation. The most effective timing for adding HMOs to improve efficacy, maximize inflammation reduction, and avoid cell toxicity was determined. The effects of 3′Sialyllactose at 1 μg / mL (FIG.5A, 5D, and 5G), neutral penta-to-hexasaccharides at 10 μg / mL (FIGS. 5B, 5E, and 5H), and 2′fucosyllactose at 10 μg / mL (FIGS.5C, 5F, and 5I) were evaluated. The results showed that under inflammatory conditions, adding the HMOs at the same time as adding the LNPs abrogated the reduction in efficacy and also reduced the inflammatory responses. Effects of HMOs with LNPs (LNP-HMOs) on mRNA delivery in mice. Studies were conducted to determine which HMO would work best when combined with LNPs in vivo via intravenous (IV) injection using the mRNA encoding luciferase. C57BL / 6 mice were IV injected with LNP-HMOs. The LNPs contained 306Oi10at 50 mol%, DSPC at 10 mol%, cholesterol at 38.5 mol%, and PEG at 1.5 mol%, and carried 0.5 mg / kg luciferase mRNA, and the amount of HMO administered was 10 µg / mouse. The HMOs tested were 3′Sialyllactose, neutral penta-to-hexasaccharides, and 2′fucosyllactose. Total luminescent flux was measured after 6 hours (FIG.6A).3′Sialyllactose had the greatest effect with this LNP in vivo and resulted in increased luciferase expression. The dose response of LNP-3′Sialyllactose was established, revealing that 10 μg / mouse had the greatest effect (FIGS.6B). Luciferase expression was increased in the liver more than in other organs (FIGS. 6C). Using different lipidoids in the LNP-HMOs with 10 μg / mouse of 3′Sialyllactose also improved LNP efficacy (FIG.6D). Taken together, these studies demonstrated that LNP-HMO improved efficacy compared to LNPs alone. Effect of human milk oligosaccharides with LNPs on mRNA delivery in mice under inflammatory conditions. To examine the kinetics of LNP-HMOs with LPS, LPS was intraperitoneally administered to C57BL / 6 mice at a dose of 0.1 mg / kg, followed by IV administration of LNP-HMOs 4 hours later. The animals underwent live imaging to determine the total luminescent flux (FIG. 7A). It was determined that 6-9 hours was the most effective time to Attorney Docket No.26100-0153WO1 2023-273 assess luciferase expression in LPS-treated mice. The dose response of 3′Sialyllactose with LNPs was then assessed in LPS-injected mice. The results showed that LNP-HMOs rescued the efficacy that was lost under inflammation. 10 μg / mouse of 3′Sialyllactose with LNPs was the most effective dose in these studies (FIGS. 7B). The increase in expression was statistically significant in the spleen LNP-HMOs (FIGS.7C). This established that under inflammatory conditions, LNP-HMOs rescued and improved the efficacy of mRNA delivery compared to LNPs alone. Example 2 The results in this Example re-present and expand on at least some of the results provided in Example 1. Screening HMOs that can reduce inflammation in murine cell lines. LNPs consist of four components: ionizable lipid, 1,2-distearoyl-sn-glycero-3- phosphocholine (DSPC), cholesterol, and PEG-lipid. For in vitro testing, the ionizable lipid 306Oi10, which is a representative, commonly used lipid for LNP delivery, was utilized at 50 mol%. DSPC was used at 10 mol%, cholesterol at 38.5 mol%, and PEG at 1.5 mol%. These LNPs were employed to deliver mRNA encoding luciferase. LNPs under inflammatory conditions exhibited reduced delivery efficacy, possibly because inflammation hindered the protein translational process (FIGS.8A and 8B). HMOs are milk sugars can have anti-inflammatory properties. To determine whether HMOs can increase the efficacy of LNPs for RNA delivery and subsequent protein translation, studies were conducted to examine whether milk sugars could reduce inflammation in the absence of LNPs. For these in vitro screens, nine oligosaccharides from human milk were tested (FIG. 9). These sugars were added to RAW-BLUETMcells and NF-κB activity was measured by QUANTI-BLUE™ Solution assay to serve as a proxy for immune activation. Initial studies determined whether HMOs themselves could stimulate the immune system (FIG. 10A).The ability of HMOs to inhibit NF-κB activity in a pre-existing inflammatory environment was then assessed using LPS-treated cells (FIG.10B). Finally, the toxicity of HMOs on the cells was measured (FIG.10C). The screening results established that the most promising HMOs for reducing inflammation were 3′Sialyllactose (3′S), neutral penta-hexasaccharides (N5 / 6), Attorney Docket No.26100-0153WO1 2023-273 and 2′Fucosyllactose (2′F).To determine whether HMOs exhibited dose responsive behavior under an inflammatory state, concentrations of HMOs ranging from 1 to 100 μg / mL were tested on both LPS-treated cells and non-inflamed (PBS treated) cells, followed by measurement of NF-κB activity. The results of these studies indicated that the most effective dose of 3′Sialyllactose was 1 μg / mL, while for neutral penta-hexasaccharides and 2′Fucosyllactose, the most effective dose was 10 μg / mL (FIGS.10D and 10E). Combining HMOs with LNPs to rescue the efficacy of mRNA delivery under inflammatory conditions in murine cell lines. LNPs consist of four components: ionizable lipid, 1,2-distearoyl-sn-glycero-3- phosphocholine (DSPC), cholesterol, and PEG-lipid. For the in vitro testing described herein, the ionizable lipid 306Oi10, representative of commonly used lipids for LNP delivery, was utilized at 50 mol%. DSPC was used at 10 mol%, cholesterol at 38.5 mol%, and PEG at 1.5 mol%. These LNPs were employed to deliver mRNA encoding luciferase. The results of these studies indicated that when combining HMOs (3′S, N5 / 6, or 2′F at 10 μg / mL) with LNPs under inflammatory conditions (LPS at 10 ng / mL), the LNP efficacy for mRNA delivery was significantly improved compared to LNPs alone under the LPS-treated condition. Additionally, the efficacy of LNPs when combined with HMOs was not significantly different than the LNPs alone without LPS treatment (FIG.10F). The timing and kinetics following the addition of 10 ng / mL LPS to cells were examined to study the efficacy of LNPs with or without the HMOs 3′S (FIG.11A) or 2′F (FIG. 11B), utilizing the ionizable lipid 200Oi10. These studies showed that adding LNPs 6 hours after LPS treatment significantly diminished LNP efficacy. Adding HMOs demonstrated the potential to rescue and / or improve LNP efficacy under inflammatory conditions. 3′Sialyllactose increased LNP efficacy under inflammatory conditions in vivo. Since HMOs demonstrated the ability to rescue LNP efficacy for mRNA delivery in pre-existing inflammatory conditions in vitro, studies were conducted to determine whether this effect would hold true in vivo. C57 / BL6 mice were injected intraperitoneally with LPS at 0.1 mg / kg. Four hours later, 306Oi10LNP combined with 3′S was intravenously injected. The Attorney Docket No.26100-0153WO1 2023-273 306Oi10LNPs encapsulated firefly luciferase mRNA at a dose of 0.5 mg / kg mRNA, and the concentration of 3′S ranged from 0.025 to 50 mg / kg (FIG.12A). The addition of 3′S was able to rescue LNP efficacy for mRNA delivery under inflammatory conditions. A dose- dependent increase in total luciferase protein expression was observed when 3′S was added with LNPs (FIG.12B). The effective concentration of 3′S was 0.5 mg / kg; above this concentration, the efficacy decreased (FIG.12C). The spleen was the major organ that contributed to the increased luciferase protein expression (FIG.12D). Cytokine and chemokine levels in the blood after LPS treatment were measured 3 hours after the administration of LNPs (FIG. 12E). Upon normalization to the LPS-only group, no significant differences were observed between the LNP groups with or without 3′S (LPS+LNP and LPS+LNP+3′S). However, the levels of all cytokines and chemokines except TNF-α and IL-10 were significantly elevated in the LNP groups (LPS+LNP and LPS+LNP+3′S) compared to LPS+3′S group . To assess the in vivo safety of HMOs, toxicity was examined through histological analysis 48 hours after the injection of LNPs. Hematoxylin and eosin (H&E) staining did not show any gross morphological changes in spleen and liver, indicating that there was no overt tissue toxicity post HMO treatment (FIG.12F). Furthermore, the LPS+LNP group showed an increase in white pulp in the spleen, suggesting an increase in the immune activity. The white pulp was reduced with LPS+LNP+3′S treatment, which aligned with the 3′S activity of reducing inflammation. 3′Sialyllactose increased LNP efficacy in vivo. To determine whether HMOs can rescue LNP efficacy for mRNA delivery under normal, non-inflamed conditions, 306Oi10LNPs encapsulating firefly luciferase mRNA were combined with 3′S and administered by intravenous injection into C57 / BL6 mice at a dose of 0.5 mg / kg of mRNA and varying concentrations of 3′S, ranging from 0.025 to 50 mg / kg (FIG.13A). A dose-dependent increase in total protein expression was observed when 3′S was co-delivered with LNPs (FIG.13B). These findings showed that 0.5 mg / kg 3′S was most effective for total protein expression. Above this concentration of 3′S, the total protein expression was decreased (FIG.13C). The liver was the major site of elevated protein expression (FIG.13D). Attorney Docket No.26100-0153WO1 2023-273 To probe systemic immunogenicity, cytokine and chemokine levels in the blood were assessed 3 hours after the administration of LNPs. Upon normalization to the untreated group, a significant decrease was observed in IL-1α levels for the LNP+3′S group compared to the LNP group. There were also significant decreases in the levels of the chemokines CCL2, CXCL1, and CXCL2 for the 3′S only group compared to the other groups (LNP and LNP+3′S) (FIG.13E). To determine whether this effect was also evident with other LNPs, the ionizable lipids SM-102 (Moderna COVID-19 vaccine ionizable lipid), DLin-MC3-DMA (ONPATTRO®), ALC-0315 (Pfizer-BioNtech COVID-19 vaccine ionizable lipid), 200Oi10, C12-200, and 503O8,12 were employed for further studies. These LNPs, encapsulating firefly luciferase mRNA, were combined with 3′S for intravenous injection into mice at 0.5 mg / kg of mRNA and 0.5 mg / kg of 3′S. In these studies, only ionizable lipid SM-102 significantly increased total protein expression when combined with 3′S. The total efficacy of the other LNPs without or with 3′S did not show a significant difference (FIG.13F). To test the effect of N5 / 6 and 2′F on LNP efficacy under normal conditions, 306Oi10LNPs encapsulating firefly luciferase mRNA were formulated with N5 / 6 and 2′F for an intravenous injection these into mice at 0.5 mg / kg of mRNA and 0.5 mg / kg of N5 / 6 or 2′F. N5 / 6 significantly decreased LNP efficacy, while 2′F maintained LNP efficacy without any improvement (FIG. 13G). Overall, the results of these studies suggested that adding HMO 3′S at a dose of 0.5 mg / kg enhanced the efficacy of LNP delivery of mRNA, particularly when using the ionizable lipids 306Oi10 and SM-102. In further studies, HMOs were added at the same time as LNPs by delivering separate solutions of the HMOs and LNPs at the same time, or alternatively, the HMOs were incorporated into LNPs in a mRNA solution, a lipid solution, or an aqueous solution. The LNPs were made up of four components: ionizable lipid, 1,2-distearoyl-sn-glycero-3- phosphocholine (DSPC) or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), cholesterol, and polyethylene glycol (PEG)-lipid. The ionizable lipid was 200Oi10, which is representative of lipids commonly used for LNP delivery. The LNPs with DSPC contained 200Oi10 at 50 mol%, DSPC at 10 mol%, cholesterol at 38.5 mol%, and PEG at 1.5 mol%. The LNPs with DOPE contained 200Oi10at 35 mol%, DSPC at 16 mol%, cholesterol at 46.5 mol%, and PEG at 2.5 mol%. The lipidoid:mRNA (w / w) ratio was 20:1 with both LNPs, Attorney Docket No.26100-0153WO1 2023-273 with 100 ng of luciferase mRNA per well. Neutral penta-to-hexasaccharides were combined at different concentrations, incorporated with the LNP, and added to RAW-BLUE™ cells. LNP efficacy was measured after 24 hours, demonstrating an increase in efficacy for the HMOs incorporated with the LNPs (FIG. 14). LNP exhibited better efficacy in the aqueous solution. OTHER EMBODIMENTS It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

Attorney Docket No.26100-0153WO1 2023-273 WHAT IS CLAIMED IS:

1. A method for delivering a therapeutic agent to a subject, wherein the method comprises administering to the subject a human milk oligosaccharide (HMO) and a lipid- containing particle comprising the therapeutic agent.

2. The method of claim 1, wherein the HMO comprises 3-Fucosyllactose, 6′Sialyllactose, 3′Sialyllactose (3′S), neutral penta-to-hexasaccharides, 2′Fucosyllactose, Sialylated Oligosaccharides, Lacto-N-fucopentaose I, Lacto-N-tetraose, or Neutral tetrasaccharides.

3. The method of claim 1, wherein said HMO comprises 3′S.

4. The method of any one of claims 1-3, wherein the HMO is administered intravenously, intramuscularly, or subcutaneously.

5. The method of any one of claims 1-4, wherein the HMO is administered at a dose of about 0.1 mg / kg to about 100 mg / kg.

6. The method of any one of claims 1-5, wherein the HMO is administered at a dose of about 0.1 mg / kg to about 50 mg / kg.

7. The method of any one of claims 1-6, wherein the lipid-containing particle is a lipid nanoparticle.

8. The method of any one of claims 1-7, wherein the lipid-containing particle is administered intravenously, intramuscularly, or subcutaneously.

9. The method of any one of claims 1-8, wherein the HMO and the lipid-containing particle are administered simultaneously.

10. The method of any one of claims 1-8, wherein the HMO and the lipid-containing particle are administered sequentially.Attorney Docket No.26100-0153WO1 2023-273 11. The method of any one of claims 1-10, wherein the therapeutic agent is a nucleic acid.

12. The method of claim 11, wherein the nucleic acid comprises an RNA.

13. The method of claim 12, wherein the RNA comprises mRNA, siRNA, shRNA, miRNA, antisense RNA, guide RNA, long non-coding RNA, transfer RNA, ribosomal RNA, dsRNA, or RNA aptamers.

14. The method of claim 12 or 13, wherein the RNA is an mRNA encoding luciferase.

15. The method of any one of claims 12-14, wherein the RNA is administered at a dose of about 0.05 mg / kg to about 5 mg / kg.

16. The method of any one of claims 12-14, wherein the RNA is administered at a dose of about 0.5 mg / kg.

17. The method of any one of claims 1-16, wherein the lipid-containing particle comprises an ionizable lipid, 1,2-distearoyl-sn-glycero-3-phosphocholine, cholesterol or a derivative thereof, and a polyethylene glycol-lipid.

18. The method of any one of claims 1-17, wherein the subject is healthy.

19. The method of any one of claims 1-18, wherein the subject has an inflammatory condition.

20. The method of claim 19, wherein the inflammatory condition comprises inflammation in a lung, liver, spleen, blood vessel, brain, lymph node, or gastrointestinal tract.

21. The method of claim 19 or 20, wherein the inflammatory condition was induced by a stimulator comprising lipopolysaccharide, lipid-containing particle, infection, injury, toxin, chronic disease, smoking, or any combination thereof.

22. The method of claim 21, wherein the stimulator is LPS or lipid-containing particles.

23. The method of any one of claims 1-22, wherein the subject is a mammal.Attorney Docket No.26100-0153WO1 2023-273 24. The method of any one of claims 1-22, wherein the subject is a mouse or a human.

25. The method of any one of claims 1-24, wherein the subject has been identified as having an infectious disease, autoimmune disease, cardiovascular disease, respiratory disease, neurological disease, cancer, metabolic disease, musculoskeletal disease, hematologic disease, or any combination thereof.

26. A composition comprising a human milk oligosaccharide (HMO) and a lipid- containing particle comprising a therapeutic agent, wherein the lipid-containing particle further comprises: a lipidoid; cholesterol or a derivative thereof; a helper lipid; and a polyethylene glycol (PEG)-based compound.

27. The composition of claim 26, wherein the HMO comprises 3-Fucosyllactose, 6′Sialyllactose, 3′Sialyllactose (3′S), neutral penta-to-hexasaccharides, 2′Fucosyllactose, Sialylated Oligosaccharides, Lacto-N-fucopentaose I, Lacto-N-tetraose, or Neutral tetrasaccharides.

28. The composition of claim 26, wherein said HMO comprises 3′S.

29. The composition of any one of claims 26-28, wherein the lipid-containing particle is a lipid nanoparticle.

30. The composition of any one of claims 26-29, wherein the therapeutic agent is a nucleic acid.

31. The composition of claim 30, wherein the nucleic acid comprises an RNA.

32. The composition of claim 31, wherein the RNA comprises one or more of a mRNA, siRNA, shRNA, miRNA, antisense RNA, guide RNA, long non-coding RNA, transfer RNA, ribosomal RNA, dsRNA, and an RNA aptamer.Attorney Docket No.26100-0153WO1 2023-273 33. The composition of claim 31 or 32, wherein the RNA is an mRNA encoding luciferase.

34. The composition of any one of claims 26-33, wherein the lipidoid is an ionizable lipid comprising one or more of 306Oi10, 200Oi10, SM-102, DLin-MC3-DMA, ALC-0315, 306O10, 503Oi10, 402O6,10, 500X1, 500Oi10; 306O11, 306O10, 306O12, 200X6, 516i10, 500O1,1,8, 514X6, 306O14, 501X1, 205O16, 500O13, 113Oi10, 306O16, 306O13, 205O18, 509X7, 501Oi10, 503Oi10, 503O8,12, 500O14, 113Oi10, 509X1, 509X3, 501X2, 402O6,10, 516O4,8, 402X8, 501O1,1,8, or 509O1,1,8.

35. The composition of any one of claims 26-34, wherein the lipidoid comprises one or more of 306Oi10, 200Oi10, SM-102, DLin-MC3-DMA, ALC-0315, C12-200, or 503O8,12.

36. The composition of any one of claims 26-35, wherein the cholesterol or a derivative thereof is cholesterol.

37. The composition of any one of claims 26-36, wherein the helper lipid is 1,2- distearoyl-sn-glycero-3-phosphocholine.

38. The composition of any one of claims 26-37, wherein the PEG-based compound is a PEG-lipid.

39. A method for treating a subject having an infectious disease, autoimmune disease, cardiovascular disease, respiratory disease, neurological disease, cancer, metabolic disease, musculoskeletal disease, hematologic disease, or any combination thereof, wherein the method comprises administering to the subject an effective amount of the composition of any one of claims 26-38.

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