Compositions and methods for modulating tumor protein p53 activity for treatment of cancer

Abstract

In certain aspects, provided herein are therapeutic compositions comprising an mRNA formulated in a lipid nanoparticle (LNP), wherein the mRNA comprises an open reading frame encoding p53 polypeptide, and methods of using the same.

Description

[0001]Attorney Docket No.7504-00302 (GRH-00562) COMPOSITIONS AND METHODS FOR MODULATING TUMOR PROTEIN p53 ACTIVITY FOR TREATMENT OF CANCER BACKGROUND Cancer impacts millions of people every year and is the second highest cause of death worldwide. All cancers mutate several tumor suppressor genes in order for the cancer to survive and thrive. The most highly mutated of these genes is TP53, which is mutated in half of all cancers. The TP53 gene's purpose is to create a protein called tumor protein p53. p53 responds to numerous cellular stresses and binds to DNA initiating repair of DNA, and if repair of the DNA is not possible, p53 induces apoptosis of the cell. Due to p53's role in the life of a cell, cancer must mutate it in order to survive. Without these mutations and a lack of functioning p53, p53 would induce apoptosis in many forms of cancer. Therefore, expressing healthy and unmutated p53 in cancer cells helps to slow cancer growth as well as induce apoptosis in cancer. SUMMARY Aspects of the invention, as provided herein, include therapeutic compositions comprising an mRNA formulated lipid nanoparticle (LNP), wherein the mRNA comprises an open reading frame encoding tumor protein o53 polypeptide (also known as cellular tumor antigen p53), or a functional fragment thereof. In some embodiments of the invention, the tumor protein p53 polypeptide, or a functional fragment thereof is cellular tumor antigen p53 isoform a transcript variant 1, cellular tumor antigen p53 isoform a transcript variant 2, cellular tumor antigen p53 isoform a transcript variant 9, cellular tumor antigen p53 isoform a transcript variant 10, cellular tumor antigen p53 isoform a transcript variant 11, cellular tumor antigen p53 isoform b transcript variant 3, cellular tumor antigen p53 isoform b transcript variant 12, cellular tumor antigen p53 isoform b transcript variant 13, cellular tumor antigen p53 isoform c, cellular tumor antigen p53 isoform d, cellular tumor antigen p53 isoform e, cellular tumor antigen p53 isoform f, cellular tumor antigen p53 isoform g transcript variant 8, cellular tumor antigen p53 isoform g transcript variant 1, cellular tumor antigen p53 isoform g transcript variant 2, cellular tumor antigen p53 isoform g transcript variant 9, cellular tumor antigen p53 isoform g transcript variant 10, cellular tumor antigen p53 isoform g transcript variant 11, cellular tumor antigen p53 isoform h, cellular tumor antigen p53 isoform i transcript variant 3, cellular tumor antigen p53 isoform i transcript variant 12, cellular tumor antigen p53 isoform i transcript variant 13, cellular tumor antigen p53 isoform, Attorney Docket No.7504-00302 (GRH-00562) cellular tumor antigen p53 isoform k, and cellular tumor antigen p53 isoform 1. Preferably, the tumor protein p53 polypeptide is cellular tumor antigen p53 isoform a transcript variant 1. In some embodiments, the LNP is a solid lipid nanoparticle (SLN). In certain aspects of the invention, provided herein are cells comprising the LNPs disclosed herein. BRIEF DESCRIPTION OF FIGURES Figure 1 depicts a DNA agarose gel representing four samples of amplified p53DNA, each having the expected length of ~1,500 base pairs relative to a 1KB DNA ladder (left-most lane). Figure 2 depicts an RNA agarose gel representing four samples of fully cleaned andcapped mRNA, each corresponding Payload 1 used to create the final lipid nanoparticle containing p53 mRNA product contemplated herein. Figure 3 depicts a single run Dynamic Light Scattering (DLS) data sheet fromcontinuous DLS data collection showing that the disclosed LNP formulations were consistently comprised of particles with a diameter of 110 nm or smaller. Figure 4 depicts an overlay of triplicate runs DLS data, illustrating the overlay of theZ-Averages (measure of the average size of a particle size distribution) and confirming minimal variance among these measurements, observed as a single visible peak. Figure 5 depicts SW1417 adenocarcinoma cells contacted with empty LNPs (control). Cells appeared as healthy cells showing normal growth. Figure 6 depicts SW1417 adenocarcinoma cells exposed to p53-mRNA-LNPs (CA-1). Tissue culture dosed with p53-mRNA-LNPs initially resulted in significantly smaller cells with some cell death and cellular debris, representing a 35-35% decrease in viability compared to control. Figure 7: depicts the p53 mRNA-LNP 24-hour cell viability assay result for SW-1417cells. Figure 8: depicts the p53 mRNA-LNP 24-hour cell total average cell count for SW-1417 cells. Figure 9: depicts the p53 mRNA-LNP 24-hour cell viability assay result for SK-N-FI cells. Figure 10: depicts the p53 mRNA-LNP 24-hour cell total average cell count for SK-N-FI cells. Attorney Docket No.7504-00302 (GRH-00562) Figure 11: depicts the p53 mRNA-LNP 48-hour (single dosing) cell viability assayresult for SW-1417 cells. Figure 12: depicts the p53 mRNA-LNP 48-hour (single dosing) cell total average cellcount for SW-1417 cells. Figure 13: depicts the p53 mRNA-LNP 48-hour (dosing every 24 hours) cell viabilityassay result for SW-1417 cells. Figure 14: depicts the p53 mRNA-LNP 48-hour (dosing every 24 hours) cell total average cell count for SW-1417 cells. Figure 15: depicts the p53 mRNA-LNP 48-hour (dosing every 24 hours) cell viability assay result for SK-N-FI cells. Figure 16: depicts the p53 mRNA-LNP 48-hour (dosing every 24 hours) cell total average cell count for SK-N-FI cells. DETAILED DESCRIPTION General In cells, p53’s purpose is to respond to genotoxic and cellular stresses and induce cell cycle arrest and apoptosis when a cell’s DNA has unrepairable DNA damage and mutations. p53 also plays a role in cellular metabolism and antioxidant response. Due to these roles, p53 plays the largest role in blocking tumor progression and the progression of cancer growth in general. Table 1p53 Sequence FeaturesMEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAA Amino Acid Sequence PPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKT CPVQLWVDSTPPPGTRVRANAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRN TFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGR DRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALEL KDAQAGKEPGGSRHSSHLKSKKGQSTSRHKKLMFKTEGPDSD (SEQ ID NO. 1)1 ctcaaaagtc tagagccacc gtccagggag caggtagctg ctgggctccg gggacactttnucleotide sequence61 gcgttcgggc tgggagcgtg ctttccacga cggtgacacg cttccctgga ttggcagcca(corresponding to Accession121 gactgccttc cgggtcactg ccatggagga gccgcagtca gatcctagcg tcgagccccc181 tctgagtcag gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtcnumber NM_00546.6181 tctgagtcag gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtc(TP53), incorporated herein301 gttcactgaa gacccaggtc cagatgaagc tcccagaatg ccagaggctg ctccccccgtby reference.)361 ggcccctgca ccagcagctc ctacaccggc ggcccctgca ccagccccct cctggcccct421 gtcatcttct gtcccttccc agaaaaccta ccagggcagc tacggtttcc gtctgggctt481 cttgcattct gggacagcca agtctgtgac ttgcacgtac tcccctgccc tcaacaagat541 gttttgccaa ctggccaaga cctgccctgt gcagctgtgg gttgattcca cacccccgcc601 oggoaccogo gtccgcgcca tggccatcta caagcagtca cagcacatga cggaggttgt661 gaggcgctgc ccccaccatg agcgctgctc agatagcgat ggtctggccc ctcctcagca721 tcttatccga gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa acacttttcg781 acatagtgtg gtggtgccct atgagccgcc tgaggttggc tctgactgta ccaccatcca841 ctacaactac atgtgtaaca gttcctgcat gggcggcatg aaccggaggc ccatcctcac901 catcatcaca ctggaagact ccagtggtaa tctactggga cggaacagct ttgaggtgcg Attorney Docket No.7504-00302 (GRH-00562)961 tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa gagaatctcc gcaagaaagg1021 ggagcctcac cacgagctgc ccccagggag cactaagcga gcactgccca acaacaccag 1081 ctcctctccc cagccaaaga agaaaccact ggatggagaa tatttcaccc ttcagatccg 1141 tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag gccttggaac tcaaggatgc 1201 ccaggctggg aaggagccag gggggagcag ggctcactcc agccacctga agtccaaaaa 1261 gggtcagtct acctcccgcc ataaaaaact catgttcaag acagaagggc ctgactcaga 1321 ctgacattct ccacttcttg ttccccactg acagcctccc acccccatct ctccctcccc1381 tgccattttg ggttttgggt ctttgaaccc ttgcttgcaa taggtgtgcg tcagaagcac1441 ccaggacttc catttgcttt gtcccggggc tccactgaac aagttggcct gcactggtgt 1501 tttgttgtgg ggaggaggat ggggagtagg acataccagc ttagatttta aggtttttac 1561 tgtgagggat gtttgggaga tgtaagaaat gttcttgcag ttaagggtta gtttacaatc 1621 agccacattc taggtagggg cccacttcac cgtactaacc agggaagctg tccctcactg 1681 ttgaattttc tctaacttca aggcccatat ctgtgaaatg ctggcatttg cacctacctc 1741 acagagtgca ttgtgagggt taatgaaata atgtacatct ggccttgaaa ccacctttta 1801 ttacatgggg tctagaactt gacccccttg agggtgcttg ttccctctcc ctgttggtcg 1861 gtgggttggt agtttctaca gttgggcagc tggttaggta gagggagttg tcaagtctct 1921 gctggcccag ccaaaccctg tctgacaacc tcttggtgaa ccttagtacc taaaaggaaa 1981 tctcacccca tcccacaccc tggaggattt catctcttgt atatgatgat ctggatccac 2041 caagacttgt tttatgctca gggtcaattt cttttttctt tttttttttt ttttttcttt 2101 ttctttgaga ctgggtctcg ctttgttgcc caggctggag tggagtggcg tgatcttggc 2161 ttactgcagc ctttgcctcc ccggctcgag cagtcctgcc tcagcctccg gagtagctgg 2221 gaccacaggt tcatgccacc atggccagcc aacttttgca tgttttgtag agatggggtc 2281 tcacagtgtt gcccaggctg gtctcaaact cctgggctca ggcgatccac ctgtctcagc 2341 ctcccagagt gctgggatta caattgtgag ccaccacgtc cagctggaag ggtcaacatc 2401 ttttacattc tgcaagcaca tctgcatttt caccccaccc ttcccctcct tctccctttt 2461 tatatcccat ttttatatcg atctcttatt ttacaataaa actttgctgc ca (SEQ ID NO. 2) In unstressed cells, there is little p53 protein expression despite mRNA for p53 being present. The E3 ligase, MDM2, targets p53 for proteasome degradation, resulting in low levels of p53 protein. In a stressed cell, inhibition or destruction of MDM2 results in the rise of p53 protein levels, which then binds as a homotetramer to several specific regulatory sequences in certain genes. There it interacts with several proteins involved in the apoptotic pathway such as NOXA, BIM, and PUMA. These proteins then induce the apoptotic pathway for a cell. In many cancer cells, p53 does not carry out its function despite the cell being stressed. This is due to several factors tha5t can occur in cancer cells: MDM2 levels / activity is unchecked due to a mutation, the p53 gene is mutated resulting in a nonfunctional p53 polypeptide, or a combination of those factors (Aubrey et al., 2017). Provided herein are nucleic acids (e.g., mRNAs) encoding tumor protein p53. The composition and methods of the present disclosure rely, at least in part, on the delivery of tumor protein p53 encoding nucleic acids (e.g., p53-encoding nucleic acids) to cells of a subject in need thereof (e.g., a gene therapy composition). For example, and without limitation, compositions comprising the tumor protein p53-encoding nucleic acids disclosed herein can be used to treat cancer. In some embodiments, said composition may be used to treat p53-associated diseases and conditions. Attorney Docket No.7504-00302 (GRH-00562) Aspects of the invention, as provided herein, include therapeutic compositions comprising an mRNA formulated in a lipid nanoparticle (LNP) (e.g., a solid lipid nanoparticle (SLN)), wherein the mRNA comprises an open reading frame encoding tumor protein p53 polypeptide, or a functional fragment thereof. In some embodiments, the tumor protein p53 polypeptide, or functional fragment thereof is selected from cellular tumor antigen p53 isoform a transcript variant 1, cellular tumor antigen p53 isoform a transcript variant 2, cellular tumor antigen p53 isoform a transcript variant 9, cellular tumor antigen p53 isoform a transcript variant 10, cellular tumor antigen p53 isoform a transcript variant 11, cellular tumor antigen p53 isoform b transcript variant 3, cellular tumor antigen p53 isoform b transcript variant 12, cellular tumor antigen p53 isoform b transcript variant 13, cellular tumor antigen p53 isoform c, cellular tumor antigen p53 isoform d, cellular tumor antigen p53 isoform e, cellular tumor antigen p53 isoform f, cellular tumor antigen p53 isoform g transcript variant 8, cellular tumor antigen p53 isoform g transcript variant 1, cellular tumor antigen p53 isoform g transcript variant 2, cellular tumor antigen p53 isoform g transcriptvariant 9, cellular tumor antigen p53 isoform g transcript variant 10, cellular tumor antigenp53 isoform g transcript variant 11, cellular tumor antigen p53 isoform h, cellular tumor antigen p53 isoform i transcript variant 3, cellular tumor antigen p53 isoform i transcript variant 12, cellular tumor antigen p53 isoform i transcript variant 13, cellular tumor antigen p53 isoform, cellular tumor antigen p53 isoform k, or cellular tumor antigen p53 isoform 1. Preferably the tumor protein p53 polypeptide, or functional fragment thereof is cellular tumor antigen p53 isoform a transcript variant 1. In some embodiments, the open reading frame is derived from the nucleic acid sequence set forth in SEQ ID NO.2, or a functional fragment thereof. For example, andwithout limitation, the mRNA comprises the nucleic acid sequence set forth in SEQ ID NO.6, SEQ ID NO. 10, or any functional fragment thereof. In some aspects, provided herein are nucleic acids encoding the p53 polypeptides disclosed herein. In certain aspects, provided herein are primers for isolating and / or amplifying a nucleic acid sequence encoding tumor protein p53 as disclosed herein, using methods known in the art (e.g., T7 RNA Polymerase techniques). In some embodiments, the primers are selected from the primer sequences set forth in SEQ ID Nos. 3, 4, 5, 7, 8, and 9.In some embodiments, the nucleic acid is isolated and / or amplified using any one of theforward and reverse primer pairs set forth in SEQ ID NOs. 3 and 5, SEQ ID NOs. 4 and 5, SEQ ID NOs. 7 and 9, and SEQ ID NOs. 8 and 9. Attorney Docket No.7504-00302 (GRH-00562) In some embodiments, one or more of the uridine nucleosides in the amplified mRNA is a pseudouridine, such as, N1-pseudouridine. In some such embodiments, all of the uridine nucleosides in the mRNA are pseudouridine, e.g., N1-methylpseudouridine. For example, and without being bound by theory or methodology, the aforementioned nucleic acids are amplified from a plasmid template. The resultant DNA sequence is used for mRNA synthesis. Without being bound by theory, and for the purpose of exemplification, the mRNAs to be incorporated in the LNPs disclosed herein may be synthesized by targeted T7 amplification of mRNA sequence from the DNA encoding p53, e.g., using primers disclosed herein. Alternatively, the mRNA is isolated from a whole cell lysate. In some embodiments, thewhole transcriptome mRNA is isolated from cell lysate. In some such embodiments cDNAsof interest, e.g., doubled-stranded cDNA encoding p53, are synthesized from the isolated mRNA, e.g., targeted amplification of sequence from forward and reverse primer pairs disclosed herein, such as in reverse transcription-polymerase chain reaction (RT-PCR). The resultant double-stranded cDNA is used for mRNA synthesis. In other aspects of the invention, provided herein are vectors comprising the nucleic acids contemplated herein. In some such embodiment, the vector is selected from nanoparticles, adenovirus vectors, adeno-associated virus (AAV) vectors, retrovirus vectors, picrona virus vectors, liposomes, cationic lipid systems, and protein / nucleic acid complexes. In some aspects of the invention, provided here are cells comprising the LNPs and / or nucleic acids disclosed herein. Preferred embodiments of the invention include cells comprising the LNPs (e.g., the SLNs) disclosed herein. In certain aspects, provided here are cells comprising the vectors disclosed herein. In further embodiments, provided herein are cells expressing the tumor protein p53 (e.g., p53 polypeptides) disclosed herein. For example and without limitation, the cell is an endothelial cell, epithelial cell, neuronal cell, or hematopoietic cell. In some such embodiments, the hematopoietic cell is an immune cell selected from a lymphocyte, a monocyte a macrophage, a dendritic cell, a mast cell, a neutrophil, a basophil, or an eosinophil. In certain embodiments, the immune cell is a T (NKT) cell a B cell, an innate lymphoid cell (ILC), a cytokine induced killer (CIK) cell, a cytotoxic T lymphocyte (CTL), a lymphokine activated killer (LAK) cell, or a regulatory T cell. In certain embodiments, the cells contemplated herein are cells of the central nervous system (CNS) or peripheral nervous system (PNS). In other embodiments, the cell is a cell of the bone marrow. Attorney Docket No.7504-00302 (GRH-00562) In some embodiments, the cell contemplated herein is a cell present in the CNS. In some embodiments, the cell is a neuronal cell. Said neuronal cell may be a sensory neuron, a motor neuron, or an interneuron. Ion other embodiments, the cell is a non-neuronal cell. In some embodiments, the non-neuronal cell is a glial cell. The glial cell may be an astrocyte cell, an oligodendrocyte, an ependymal cell, a radial glial cell, a Schwann cell, a satellite cell, an enteric glial cell, or a microglial cell. In some embodiments, the nucleic acids contemplated herein may refer to a polymeric form of nucleotides or nucleosides of any length, such as deoxyribonucleotides or ribonucleotides, or analogs thereof. Nucleic acids may have any three-dimensional structure, and may perform any function. The following are non-limiting examples of nucleic acids: coding or non-coding regions of a gene or gene fragment, loci (locus) defined from linkage analysis, exons, introns, messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozymes, cDNA, recombinant polynucleotides / polynucleosides, branched polynucleotides / polynucleosides, plasmids, vectors, isolated DNA of any sequence, isolated RNA of any sequence, nucleic acid probes, and primers. A nucleic acid may comprise modified nucleotides / nucleosides, such as methylated nucleotides / nucleosides and nucleotide / nucleoside analogs. If present, modifications to the polynucleotide / polynucleoside structure may be imparted before or after assembly of the polymer. A polynucleotide / polynucleoside may be further modified, such as by conjugation with a labeling component. Aspects of the invention include therapeutic compositions comprising the mRNAs disclosed herein. In some embodiments, the therapeutic composition comprises a vectorselected from nanoparticles, adenovirus vectors, adeno-associated virus (AAV) vectors,retrovirus vectors, picorna virus vectors, liposomes, cationic lipid systems, and protein / nucleic acid complexes. For example and without limitation, the therapeutic composition comprising an mRNA may be formulated in a lipid nanoparticle (LNP). In some embodiments of the therapeutic composition, one or more of the uridine nucleosides in the mRNA are pseudouridine, such as, N1-methyl pseudouridine. In some embodiments, all of the uridine nucleosides in the mRNA are pseudouridine, e.g., N1-methylpseudouridine. In some embodiments of the therapeutic composition, the LNP (e.g., the SLN) comprises an ionizable lipid, a structural lipid, a phospholipid, a sterol, a PEG-modified lipid, or any combination thereof. In some aspects, provided herein are methods of treating a cancer in a subject, the method comprising administering an effective amount of a therapeutic composition Attorney Docket No.7504-00302 (GRH-00562) contemplated herein. In some embodiments, the cancer is selected from: hepatocellular carcinoma, lymphoma, B cell lymphoma, T cell lymphoma, mycosis fungoides, Hodkin’s Disease, myeloid leukemia, bladder cancer, brain cancer, nervous system cancer, head and neck cancer, squamous cell carcinomas of the mouth, throat, larynx, and lung, endometrial cancer, cervical cancer, cervical carcinoma, breast cancer, epithelial cancer, renal cancer, genitourinary cancer, pulmonary cancer, esophageal carcinoma, head and neck carcinoma, large bowel cancer, hematopoietic cancers, testicular cancer, colon and rectal cancer, prostatic cancer, and pancreatic cancer. In some embodiments, the therapeutic composition is administered intrapleurally, intravenously, subcutaneously, intranodally, intraumorally, intrathecally, intraperitoneally, intracranially, or by direct administration to an organ. In certain embodiments, the method further comprises administering to the subject an immunotherapy. The immunotherapy may comprise administration of a therapeutic antibody such as monoclonal antibodies, including rituximab, cetuximab, bevacizumab, pertuzumab, and the like. The immunotherapy may comprise administration of an immune checkpoint inhibitor. In some such embodiments, the immune checkpoint inhibitor comprises an antibody or antigen-binding fragment thereof specific for PD-1, PD-L1, or CTLA4, such as nivolumab and ipilimumab. In certain embodiments, cancer immunotherapy comprises administration of a CAR-T cell or a CAR-NK cell. In other embodiments, the method further comprises administering to the subject chemotherapy, radiation, immunosuppressive agents, or performance of surgery, as are well- known in the art for the treatment of cancers and tumors. Definitions For convenience, certain terms employed in the specification, examples, and appended claims are collected here. The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. The term “about” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. Where the terms “about” or “approximately” are used in the context of compositions containing amounts of ingredients or conditions such as temperature, these values include the state value with a variation of 0-10% around the value (X ± 10%). Attorney Docket No.7504-00302 (GRH-00562) Ranges are stated in the shorthand to avoid having to set out at length and describe each and every value within the range. Therefore, when ranges are stated for a value, any appropriate value of the range. For example, a range of 0.1-1.0 represents the terminal values of 0.1 and 1.0, as well as the intermediate values of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, and allintermediate ranges encompassed within 0.1-1.0, such as 0.2-0.5, 0.2-0.8, 0.7-1.0, etc.“Pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient”, includes any and all solvents, dispersion media, coating, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with angiotensin II, its use in the pharmaceutical formulations of the invention is contemplated. In certain embodiments, the pharmaceutically acceptable canier / excipient is a saline solution. As used herein, the term “administering” means providing a pharmaceutical agent or composition to a subject, and includes, but is not limited to, administering by a medical professional and self-administering. Such an agent can contain, for example, peptide or nucleic acid described herein. As used herein, the term “treatment” refers to clinical intervention designed to alter the natural course of the individual being treated during the course of clinical pathology. Desirable effects of treatment include decreasing the rate of progression, ameliorating or palliating the pathological state, and remission or improved prognosis of a particular disease, disorder, or condition. An individual is successfully “treated”, for example, if one or more symptoms associated with a particular disease, disorder, or condition are mitigated or eliminated. As used herein, a therapeutic that “prevents” a condition refers to a compound that, when administered to a statistical sample prior to the onset of the disorder or condition, reduces the occurrence of the disorder or condition in the treated sample relative to an untreated control sample, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition relative to the untreated control sample. In certain embodiments, agents of the invention may be used alone or conjointly administered with another type of therapeutic agent. As used herein, the phrase “conjoint administration” or “administered conjointly” refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., the two agents are simultaneously effective in the subject, which may include synergistic effects of the two Attorney Docket No.7504-00302 (GRH-00562) agents). For example, the different therapeutic compositions disclosed herein can be administered either in the same formulation or in separate formulations, either concomitantly or sequentially. In certain embodiments, the different therapeutic agents (e.g., a therapeutic composition comprising an mRNA disclosed herein and an immunotherapy or standard-of- care treatment (e.g., standard-of-care treatment for a cancer, such as Adenocarcinoma of the Colon)) can be administered within about one hour, about 12 hours, about 24 hours, about 36 hours, about 48 hours, about 72 hours, or about a week of one another. Thus, a subject who receives such treatment can benefit from a combined effect of different therapeutic agents. The terms “polypeptide fragment” or “fragment”, when used in reference toa particular polypeptide, refers to a polypeptide in which amino acid residues are deleted as compared to the reference polypeptide itself, but where the remaining amino acid sequence is usually identical to that of the reference polypeptide. Such deletions may occur at the amino- terminus or carboxy-terminus of the reference polypeptide, or alternatively both. Fragments typically are at least about 5, 6, 8, or 10 amino acids long, at least about 14 amino acids long, at least 20, 30, 40 or 50 amino acids long, at least about 75 amino acids long, or at least about 100, 150, 200, 300, 500 or more amino acids long. A fragment can retain one or more of the biological activities of the reference polypeptide. In various embodiments, a fragment may comprise an enzymatic activity and / or an interaction site of the reference polypeptide. In some embodiments, a fragment may have suppressive, disruptive, or enhancing properties. Nucleic acids and vectors Nucleic acids and vectors disclosed herein include polynucleotides and polynucleotide vectors encoding the disclosed tumor protein p53 (e.g., p53 polypeptides) that allow expression in the disclosed cells. Nucleic acid sequences contemplated herein can be obtained using recombinant methods known in the art. Alternatively, the sequence of interest can be produced synthetically, rather than cloned. In addition to the polypeptide-encoding sequences, other structural properties as described herein for mRNA constructs (e.g., modified nucleobases, 5’ cap, 5’ UTR, 3’ UTR, miR binding site(s), polyA tail, as described herein). Suitable mRNA construct components arc as described herein. In some embodiments, a nucleic acid of the disclosure may be modified in a coding region (e.g., an open reading frame of an mRNA encoding polypeptide). In other embodiments, nucleic acid may be modified in regions besides a coding region, such as 5’ cap, a 5’-untranslated region (UTR) and / or a 3’-UTR, polyA tail of an mRNA, wherein any combination of elements may be independently modified. In some embodiments, such Attorney Docket No.7504-00302 (GRH-00562) regions may contain one or more different nucleoside modifications. I such embodiments, modifications may also be present in the coding region. Examples of nucleoside modifications and combinations thereof that may be present in mRNAs disclosed herein include, but are not limited to, those described in PCT PatentApplication Publications: WO 2012 / 045075, WO 2014 / 081507, WO 2014 / 093924, WO2014 / 164253, WO 2014 / 159813, WO 2018 / 144775, WO 2018 / 081459, each of which are incorporated herein in their entirety. In some embodiments, the mRNAs of the disclosure can include a combination of modifications to the sugar, the nucleobase, and / or the internucleoside linkage. These combinations can include any one of more modifications described herein. As a non-limiting example, the natural nucleotide uridine may be substituted with a modified nucleoside described herein. In another non-limiting example, the natural nucleoside uridine may be partially substituted (e.g., about 0.1%, 1%. 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99.9% of the natural uridines) with at least one of the modified nucleosides disclosed herein, e.g., pseudouridine. Expression of nucleic acids encoding tumor protein p53 (e.g., p53) is typically achieved by operably linking a nucleic acid encoding the p53 polypeptide to a promoter, and incorporating the construct into an expression vector. Typical cloning vectors contain transcription and translation terminators, initiation sequences, and promoters useful for regulation of the expression of the desired nucleic acid sequence. The disclosed nucleic acids can be cloned into a number of types of vectors. For example, the nucleic acid can be cloned into a vector including, but not limited to a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors, and sequencing vectors. Further, the expression vector may be provided to a cell in the form of a viral vector. Viral vector technology is well known in the art and is described, for example, in Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and in other virology and molecular biology manuals. Viruses, which are useful as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, and lentiviruses. In general, a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers. In some embodiments, the polynucleotide vectors are lentiviral or retroviral vectors. Attorney Docket No.7504-00302 (GRH-00562) A number of viral based systems have been developed for gene transfer intomammalian cells. For example, retroviruses and AAVs provide convenient platform for genedelivery systems. A selected gene can be inserted into a vector and packaged in viral particles using techniques known in the art. The recombinant virus can then be isolated and deliveredto cells of the subject either in vivo or ex vivo.One example of a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence capable of driving high levels of expression of any polynucleotide sequence operativelylinked thereto. Another example of a suitable promoter is Elongation Growth Factor- -limited to the simian virus 40 (SV40) early promoter, MND (myeloproliferative sarcoma virus) promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus (HIV) long terminal repeat (LTR) promoter, MoMuLV promoter, an avian leukemia virus promoter, an Epstein-Barr virus immediate early promoter, a Rous sarcoma virus promoter, as well as human gene promoters such as, but not limited to, the actin promoter, the myosin promoter, the hemoglobin promoter, and the creatine kinase promoter. The promoter can alternatively be an inducible promoter. Examples of inducible promoters include, but are not limited to a metallothionine promoter, a glucocorticoid promoter, a progesterone promoter, and a tetracycline promoter. Additional promoter elements, e.g., enhancers, regulate the frequency of transcriptional initiation. Typically, these are located in the region 30-110 base pairs (bp) upstream of the start site, although a number of promoters have recently been shown to contain functional elements downstream of the start site as well. The spacing between promoter elements frequently is flexible, so that promoter function is preserved when elements are inverted or more relative to one another. In order to assess the expression of tumor protein p53 disclosed herein or portions thereof, the expression vector to be introduced into a cell can also contain either a selectable marker gene or a reporter gene or both to facilitate identification and selection of expression expressing cells from the population of cells sought to be transfected or infected through viral vectors. The selectable marker may be carried on a separate piece of DNA and used in a co- transfection procedure. Both selectable markers and reporter genes may be flanked with appropriate regulatory sequences to enable expression in the host cells. Useful selectable markers include, for example, antibiotic-resistance genes. Attorney Docket No.7504-00302 (GRH-00562) Reporter genes may be used for identifying potentially transfected cells and for evaluating the functionality of regulatory sequences. In general, a reporter gene is a gene that is not present in or expressed by the recipient organism or tissue and that encodes a polypeptide whose expression is manifested by some easily detectable property, e.g.,enzymatic activity. Expression of the reporter gene is assayed at a suitable time after thenucleic acid has been introduced into the recipient cells. Suitable reporter genes may include genes encoding luciferase, beta-galactosidase, chloramphenicol acetyl transferase, secreted alkaline phosphatase, or the green fluorescent protein gene. Suitable expression systems are well known and may be prepared using known techniques or obtained commercially. Ingeneral, the construct with the minimal 5’ flanking region showing the highest level ofexpression of reporter gene is identified as the promoter. Such promoter regions may be linked to a reporter gene and used to evaluate agents for the ability to modulate promoter- driven transcription. Methods of introducing and expressing genes into a cell are known in the art. In the context of an expression vector, the vector can be readily introduced into a host cell, e.g., mammalian, bacterial, yeast, or insect cell by any method in the art. For example, the expression vector can be transferred into a host cell by physical, chemical, or biological means. Physical methods for introducing a polynucleotide into a host cell include calcium phosphate precipitation, lipofection, particle bombardment, microinjection, electroporation, and the like. Methods for producing cells comprising vectors and / or exogenous nucleic acids are well-known in the art. See, for example, Sambrook et al. (2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). Biological methods for introducing a polynucleotide of interest into a host cell include the use of DNA and RNA vectors. Viral vectors, and especially retroviral vectors, have become the most widely used method for inserting genes into mammalian, e.g., human cells. Chemical means for introducing a polynucleotide into a host cell include colloidal dispersion systems, such as macromolecular complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system for use as a delivery vehicle in vitro and in vivo is a liposome (e.g., an artificial membrane vesicle). In the case where a non-viral delivery system is utilized, an exemplary delivery vehicle is a liposome. In another aspect, the nucleic acid may be associated with a lipid. The nucleic acid associated with a lipid may be encapsulated in the aqueous interior of a Attorney Docket No.7504-00302 (GRH-00562) liposome, interspersed within the lipid bilayer of a liposome, attached to a liposome via alinking molecule that is associated with both the liposome and the oligonucleotide, entrappedin a liposome, complexed with a liposome, dispersed in a solution containing a lipid, mixed with a lipid, combined with a lipid, contained as a suspension in a lipid, contained or complexed with a micelle, or otherwise associated with a lipid. Lipid, lipid / nucleic acid orlipid / expression vector associated compositions are not limited to any particular structure insolution. For example, they may be present in a bilayer structure, as micelles, or with a“collapsed” structure. They may also simply be interspersed in a solution, possibly formingaggregates that are not uniform in size or shape. Lipids are fatty substances, which may be naturally occurring or synthetic lipids. For example, lipids include the fatty droplets that naturally occur in the cytoplasm as well as the class of compounds, which contain long-chainaliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, aminoalcohols, and aldehydes. Lipids suitable for use can be obtained from commercial sources. For example, dimyristyl phosphatidylcholine (“DMPC”) can be obtained from Sigma, St. Louis, Mo.; dicetyl phosphate (“DCP”) can be obtained from K & K Laboratories (Plainview, N.Y.); cholesterol (“Choi”) can be obtained from Calbiochem-Behring; dimyristyl phosphatidylglycerol (“DMPG”) and other lipids may be obtained from Avanti Polar Lipids, mc, (Birmingham, Ala.). In some embodiments the nucleic acids of the disclosure may be formulated in nanoparticles (e.g., lipid nanoparticles) or other delivery vehicles, e.g., to protect them from degradation when delivered to a subject. Illustrative nanoparticles are described in Panyam, J. & Labhasetwar, V. Adv. Drug Deliv. Rev. 55, 329-347 (2003) and Peer, D. et al. Nature Nanotech. 2, 751-760 (2007), WO 2018 / 144775, and WO 2018 / 081459, each of which are incorporated herein by reference in their entirety. In certain embodiments, an mRNA of the disclosure is encapsulated within a nanoparticle. In particular embodiments, a nanoparticle is a particle having at lease one dimension (e.g., a diameter) less than or equal to 1000 nanometers, less than or equal to 500 nm or less than or equal to 100 nm. In particular embodiments, a nanoparticle includes lipids. Lipid nanoparticles (LNPs) include, but are not limited to, solid lipid nanoparticles (SLNs), liposomes, and micelles. For example, and without limitation, the nucleic acids described herein (e.g., mRNAs) are formulated as a solid lipid nanoparticle (SLN), which can be spherical with an average diameter between 10 to 1000 nm. In some such embodiments, the SLN possess a solid lipid core matrix that can solubilize lipophilic molecules and can be stabilized with surfactants and / or emulsifiers, Attorney Docket No.7504-00302 (GRH-00562) Exemplary SLN can be those as described in Inti. Pub. No. WO 2013 / 105101, herein incorporated by reference in its entirety. Any of a number of lipids may be present, including cationic and / or ionizable lipids, anionic lipids, neutral lipids, amphipathic lipids, PEGylated lipids, and / or structural lipids. Such lipids can be used alone or in combination. In certain embodiments, a lipid nanoparticle comprises one or more nucleic acids, e.g., mRNAs, described herein. In certain embodiments, it is desirable to target a nanoparticle, e.g., a lipid nanoparticle, of the disclosure using a targeting moiety that is specific to a cell type and / or tissue type. In some embodiments, a nanoparticle may be targeted to a particular cell, tissue, and / or organ using a targeting moiety. In particular embodiments, a nanoparticle comprises one or more mRNA described herein and a targeting moiety. Exemplary non-limiting targeting moieties include ligands, cell surface receptors, glycoproteins, vitamins (e.g., riboflavin) and antibodies (e.g., full-length antibodies, antibody fragments (e.g., Fv fragments, single chain Fv (scFv) fragments, Fab’ fragments, or F(ab’)2 fragments), single domain antibodies, camelid antibodies, and multispecific antibodies (e.g., bispecific antibodies)). In some embodiments, the targetingmoiety may be a polypeptide. The targeting moiety may include the entire polypeptide (e.g.,peptide or protein) or fragments thereof. A targeting moiety is typically positioned on theouter surface of the nanoparticle in such a manner that the targeting moiety is available forinteraction with the target, for example, a cell surface receptor. A variety of different targetingmoieties and methods are known and available in the art, including those described, e.g., inSapra et al., Prog. Lipid Res. 42(5):439-62, 2003 and Abra et al., J. Liposome Res. 12:1-3,2002. For example, the lipid nanoparticle may include a targeting moiety that targets the lipidnanoparticle to a cell including, but not limited to, hepatocytes, colon cells, epithelial cells,hematopoietic cells, epithelial cells, endothelial cells, lung cells, bone cells, stem cells,mesenchymal cells, neural cells, cardiac cells, adipocytes, vascular smooth muscle cells,cardiomyocytes, skeletal muscle cells, beta cells, pituitary cells, synovial lining cells, ovariancells, testicular cells, fibroblasts, B cells, T cells, reticulocytes, leukocytes, granulocytes, andtumor cells (including primary tumor cells and metastatic tumor cells). In particular embodiments, the targeting moiety targets the lipid nanoparticle to a hepatocyte. In other embodiments, the targeting moiety targets the lipid nanoparticle to a colon cell. In some embodiments, the targeting moiety targets the lipid nanoparticle to a liver cancer cell (e.g., a hepatocellular carcinoma cell) or a colorectal cancer cell (e.g., a primary tumor or a metastasis). Attorney Docket No.7504-00302 (GRH-00562) In addition to nanoparticles compositions provided herein, also disclosed are methodsof producing lipid nanoparticles, which may include encapsulating a polynucleotide (e.g., anmRNA contemplated herein). Such contemplated methods comprise using any of the compositions disclosed herein and producing lipid nanoparticles in accordance with methodsof production of lipid nanoparticles known in the art, e.g., Wang et al. (2015) “Delivery ofoligonucleotides with lipid nanoparticles” Adv. Drug Deliv. Rev. 87:68-80; Silva et al. (2015) “Delivery Systems for Biopharmaceuticals. Part I: Nanoparticles and Microparticles” Curr. Pharm. Biotechnol. 16: 940-954; Naseri et al. (2015) “Solid Lipid Nanoparticles and Nanostructured Lipid Carriers: Structure, Preparation and Application” Adv. Pharm. Bull. 5:305-13; Silva et al. (2015) “Lipid nanoparticles for the delivery of biopharmaceuticals” Curr. Pharm. Biotechnol. 16:29 1-302, and references cited therein, all of which are incorporated herein by reference in their entirety. In certain embodiments, lipid nanoparticles (LNPs) comprise lipids including an ionizable lipid, a structural lipid, a phospholipid, astabilizing lipid, and one or more mRNAs. For example, without being bound by theory ormethodology, a solid lipid nanoparticle (SLN) may include one or more mRNAs. Thus, each if the LNPs described herein may be used in a formulation comprising the mRNA described herein. In one embodiment, a lipid nanoparticle comprises an ionizable lipid, a structural lipid, a phospholipid, a PEG-modified lipid, a sterol and a phospholipid. In some embodiments, the LNP has a molar ratio about 20-60% ionizable lipid: about 5-25% phospholipid: about 25-55% sterol: and about 0.5-15% PEG-modified lipid, about 38.5% cholesterol and about 10% phospholipid. In some embodiments, the LNP comprises a molar ratio of about 55% ionizable lipid, about 2.5% PEG lipid, about 32.5% cholesterol and about 10% phospholipid. In some embodiments, the ionizable lipid is an ionizable amino or cationic lipid and the neutral lipid is a phospholipid, and the sterol is a cholesterol. The ionizable lipids include but are not limited to, SM-102, 9-Heptadecanyl 8-{(2- hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino}octanoate, 3-(didodecylamino)-N1,N4,N4- tridodecyl-1-piperazineethanamine (KL10), N1-[2-(didodecylamino)ethyl]-N1,N4,N4- tridoceyl-1,4,-piperazinediethanamine (KL22), 14,25-ditridecyl-15,18,21,24-tetraaza-octatriacontane (KL25), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (Dlin-DMA), 2,2-dilinoleyl-4-(dimethylaminomethyl-[1,3]-dioxolane (Dlin-K-DMA), heptatriaconta- 6,9,28,31-tetraen-19-yl 4-(dimethylamino)butanoate (Dlin-MC3-DMA), 2,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (Dlin-KC2-DMA), 2-({8- -cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9z,12Z)-octadeca-9,12-dien-1-yloxy]propan-1-amine(Octyl-CLinDMA), (2R)-2-({8- -cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3- Attorney Docket No.7504-00302 (GRH-00562)[(9Z,12Z)-octadeca-9, 12-dien-1-yloxyl]propan-(Octyl-CLinDMA (2R)), (2S)-2-({8- -cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-9,12-dien-1-yloxypropan-1- amine (Octyl-CLinDMA (2S)). N,N-dioleyl-N,N-dimethylammonium chloride (“DODAC”): N-(2,3-dioleyloxy)propyl-N,N--N-triethylammonium chloride (“DOTMA”); N,N-disterayl- N,N-dimethylammonium bromide (“DDAB”); N-(2,3-dioleoyloxy)propyl)-N,N,N- trimethylammonium chloride (“DOTAP”); 1,2-Dioleyloxy-3-trimethylarninopropane chloridesalt (“DOTAP.01”); 3- -(N-(N’,N’-dimehtylaminoethane)-carbamoyl)cholesterol (“DC-Chol”), N-(1-(2,3-diolexyloxy)propyl)-N-2-(sperminecarboxamido)ethyl)-N,N-dimethyl- ammonium trifluoraceate (“DOSPA”), dioctadecylamidoglycyl carboxyspermine (“DOGS”), 1,2-dioleoyl-3dimethylammonium propane (“DODAP”), N,N-dimethyl-2,3- dioleyloxy)propylamine (“DODMA”), and N-(1,2-dimyristyloxyprop-3-yl)-N,N-dimethyl-N- hydroxyethyl ammonium bromide (“DMRIE”). Additionally, a number of commercial preparations of cationic and / or ionizable lipids can be used, such as, e.g., LIPIOFECTIN® (including DOTMA and DOPE, available from GIBCO / BRL), and LIPOFECTAMINE® (including DOSPA and DOPE, available from GIBCO / BRL). KL10, KL22, and KL25 are described, for example, in U.S. Pat. No, 8,691,750, which is incorporated herein by reference in its entirety. In particular embodiments, the lipid is Dlin-MC3-DMA, Dlin-KC2-DMA, or ALC-0159. The phospholipids provided herein may, for example, be one or more saturated or (poly)unsaturated phospholipids or a combination thereof. In general, phospholipids comprise a phospholipid moiety and one or more fatty acid moieties. A phospholipid moiety can be selected, for example, from the non-limiting group consisting of phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, phosphatidyl serine, phosphatidic acid, 2-lysophosphatidyl chole, and a sphingomyelin. A fatty acid moiety can be selected, for example, from the non-limiting groupconsisting of lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid,stearic acid, oleic acid, linoleic acid, alpha-linolenic acid, erucic acid, phytanoic acid,arachidic acid, arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid,and docosahexaenoic acid. Particular phospholipids can facilitate fusion to a membrane. For example, a cationic phospholipid can interact with one or more negatively charged phospholipids of a membrane (e.g., a cellular or intracellular membrane). Fusion of a phospholipid to a membrane can allow one or more elements (e.g., a therapeutic agent) of a lipid-containing composition (e.g., Attorney Docket No.7504-00302 (GRH-00562) LNPs) to pass through the membrane permitting, e.g., delivery of the one or more elements to a target tissue. Non-natural phospholipid species including natural species with modifications and substitutions including branches, oxidation, cyclization, alkynes are also contemplated. For example, a phospholipid can be functionalized with or cross-linked to one or more alkynes (e.g., an alkenyl group in which one or more double bonds is replaced with a triple bond). Under appropriate reaction conditions, an alkyne group can undergo a copper-catalyzed cycloaddition upon exposure to an azide. Such reactions can be useful in functionalizing the surface (e.g., the lipid monolayer or bilayer) of a nanoparticle composition to a useful component such as a targeting or imaging moiety (e.g., a dye). Phospholipids include, but are not limited to, glycerophospholipids such as phosphatidylcholines, phosphatidylethanolamines, phosphatidylserines, phosphatidylinositols, phosphatidylglycerols, and phosphatidic acids. In some embodiments, the phospholipid is distearoylphosphatidylcholine (DSPC). Phospholipids also include phosphosphingolipid, such as sphingomyelin. The lipid composition of a pharmaceutical composition disclosed herein can comprise one or more structural lipids. As used herein, the term “structural lipid” refers to sterols and also to lipids containing sterol moieties. Incorporation of structural lipids in the lipid nanoparticle may help aggregation of other lipids in the particle. Structural lipids can be selected from the group including but not limited to, cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, tomatine, ursolic acid, alpha-tocopherol, hopanoids, phytosterols, steroids, and mixtures thereof. In some embodiments, the structural lipid is a sterol. As defined herein, “sterols” are a subgroup of steroids consisting of steroid alcohols. In certain embodiments, the structural lipid is a steroid. In some embodiments, the structural lipid is cholesterol. In certain embodiments, the structural lipid is an analog of cholesterol. The term “PEG-modified lipid” may refer to polyethylene glycol (PEG)-modified lipids. Non-limiting examples of PEG-lipids include PEG-modified phosphatidylethanolamine and phosphatidic acid, PEG-ceramide conjugates (e.g., PEG- CerC14 or PEG-CerC20), PEG-modified dialkylamines and PEG-modified 1,2- diacyloxpropan-3-amines. Such lipids are also referred to as PEGylated lipids. For example, a PEG lipid can be PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, or a PEG-DSPE lipid. Attorney Docket No.7504-00302 (GRH-00562) In some embodiments, the PEG-lipid includes, but is not limited to 1,2-dimyrstoyl-sn- glycerol methoxypolyethyleneglycol (PEG-DMG), 1,2-diasteroyl-sn-glycero-3- phosphoethanolamine-N-[amino(polyethylene glycol)] (PEG-DSPE), PEG-disteryl glycerol (PEG-DSG), PEG-dipalmitoyl phosphatidylethanolamine (PEG-DPPE), or PEG-1,2- dimyrstyloxlpropyl-3-amine (PEG-c-DMA). Preferably, the PEG-modified lipid is 1,2- dimyristoyl-sn-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG 2000). In some embodiments, the PEG-lipid is selected from a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, and mixture thereof. In some embodiments, the lipid moiety of the PEG-lipids includes those having lengths of from about C14 to about C22, preferably from about C14 to about C16. In some embodiments, a PEG moiety, for example an mPEG-NH2, has a size of about 1000, 2000, 5000, 10,000, 15,000 or 20,000 daltons. In some embodiments, the PEG-lipid is PEG2k- DMG. In certain embodiments, the lipid nanoparticles described herein can comprise a PEG lipid which is a non-diffusible PEG. Non-limiting examples of non-diffusible PEGs include PEG-DSG and PEG-DSPE PEG-lipids are known in the art, such as those described in U.S. Pat. No, 8,158,601 and International Publ. No. WO 2015 / 130584 A2, which are incorporated herein by reference in their entirety. In general, some of the other lipid components (e.g., PEG lipids) of various formulae, described herein may be synthesized as described in PCT Application WO 2017 / 099823 entitled “Compositions and methods for Deliver of Therapeutic Agents,” which is now incorporated herein by reference in its entirety. The lipid component of a lipid nanoparticle composition may include one or more molecules comprising polyethylene glycol, such as PEG or PEG-modified lipids. Such species may be alternatively referred to as PEGylated lipids. A PEG lipid is a lipid modified with polyethylene glycol. A PEG lipid may be selected from the non-limiting group including PEG-modified phosphatidylethanolamines, PEG-modified phosphatidic acids, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols, and mixtures thereof. For example, a PEG lipid may be DMG-PEG 2000, PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, or a PEG-DSPE lipid. Attorney Docket No.7504-00302 (GRH-00562) In some embodiments the PEG-modified lipids are a modified form of PEG DMG, including DMG-PEG 2000. In some embodiments, the LNPs comprise ALC-0159, a PEGylated lipid; the N,N- dimyristylamide of 2-hydroxyacetic acid, O-pegylated to a PEG chain mass of about 2 kilodaltons. In certain embodiments, the LNPs comprise ALC-0315, a synthetic ionizable cationic amino lipid. “Stabilizing lipids”, as used herein, may include, but is not limited to, lipids that contain surface stabilizing polymers conjugated to the lipid headgroup. In some embodiments, the polymer conjugated to the lipid headgroup is hydrophilic. The hydrophilic polymer-conjugated lipid may be a polyethyleneglycol (PEG)-conjugated lipid. In other embodiments, the polymer making up the polymer-lipid conjugate can be a polymer that contains a backbone that allows it to associate with the core of the particle thereby enhancing the stability of the delivery vehicle (e.g., poly(vinyl alcohol) conjugated to a lipid). PEG lipids may be used to stabilize the nanoparticle, e.g., in terms of making it invisible to the immune system. Without being bound by theory, hydrophilic polymer PEG on the outer surface of the nanoparticle induces steric stabilization due to the local surface concentration of highly hydrated PEG groups. This attracts a water shell that surrounds the nanoparticle that acts as a barrier against certain interactions in the biological environment, e.g., making the nanoparticle less detectable by, or otherwise invisible to, the immune system, including inhibition of adsorption and opsonization of the nanoparticle and its contents. Such nanoparticles may have reduced detection and destruction in the biological environment, and can lead to extended blood circulation time and preferential accumulation at target sites. Stabilizing lipids may include some lipids that are not conjugated to a stabilizing polymer. Such lipids contain a negatively charged phosphate group shielded by a hydrophilic neutral moiety such as phosphatidylglycerol (PG) and phosphatidylinositol (PI). In some embodiments, the LNP has a molar ratio of 50:38.5:10:1.5 of ionizable lipid: structural lipid: phospholipid: PEG-modified lipid. Preferably the LNP has a molar ratio of 50:10:38.5:1.5 of SM-102: cholesterol: DSPC (Distearoylphosphatidylcholine): DMG-PEG 2000. In some such embodiments, the LNP is a solid lipid nanoparticle (SLN). Compositions In some aspects, provided herein are compositions (e.g., a pharmaceutical composition, such as a therapeutic or vaccine composition), comprising the nucleic acids disclosed herein, optionally formulated together with a pharmaceutically acceptable carrier, Attorney Docket No.7504-00302 (GRH-00562) (e.g., a composition comprising the nanoparticles disclosed herein) as well as methods of administering such pharmaceutical compositions. In some embodiments, the nucleic acids, polypeptides, or compositions provided herein are used as an adjuvant. As used, herein, the term “adjuvant” broadly refers to an agent that affects an immunological or physiological response in a patient or subject. For example and without limitation, when used as an adjuvant the polypeptides or compositions provided herein may increase the presence of an antigen over time or to an area of interest like a tumor, facilitate absorption of a presented antigen, activate macrophages and lymphocytes, and / or support the production of cytokines. By changing an immune response, the adjuvant might permit a smaller dose of an immune interacting agent to increase the effectiveness or safety of a particular dose of the immune interacting agent. For example, the adjuvant might prevent T cell exhaustion and thus increase the effectiveness or safety of a particular immune interacting agent. Compositions contemplated herein may be administered intrapleurally, intravenously, subcutaneously, intranodally, intratumorally, intrathecally, intraperitoneally, intracranially, or by direct administration to an organ. Said compositions may comprise one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solution, dispersions,suspensions, or emulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending thickening agents. Examples of suitable aqueous and nonaqueous carriers which may be employed in the pharmaceutical compositions include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof vegetable oils, such as olive oil, and injectable organic esters, such as ethyloleate. Proper fluidity can be maintained, for example, by is of the coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. In some embodiments, the administered dose size is about 100 μL to about 50 mL (50,000 μL) or any intermediate value encompassed therein, particularly, about: 100 μL, 200μL, 300 μL, 400 μL, 500 μL, 600 μL, 700 μL, 800 μL, 900 μL, or 1000 μL (1 mL), 2 mL, 3mL, 4 mL, 5 mL, 6 mL, 7 mL, 8 mL, 9 mL, 10 mL, 11 mL, 12 mL, 13 mL, 14 mL, 15 mL, 16 mL, 17 mL, 18 mL, 19 mL, 20 mL, 21 mL, 22 mL, 23 mL, 24 mL, 25 mL, 26 mL, 27 mL, 28 mL, 29 mL, 30 mL, 31 mL, 32 mL, 33 mL, 34 mL, 35 mL, 36 mL, 37 mL, 38 mL, 39 mL, 40 mL, 41 mL, 42 mL, 43 mL, 44 mL, 45 mL, 46 mL, 47 mL, 48 mL, 49 mL, or 50 mL. In some Attorney Docket No.7504-00302 (GRH-00562)preferred embodiments, the dose size is 10 mL. In some such embodiments, the doseencompassed therein, particularly, about: ,, or 2,500 In some such embodiments, the doseadministered may be at a concentration of about 0.002 mg / kg to about 0.03 mg / kg mRNA or any intermediate value encompassed therein such as about: 0.003 mg / kg, 0.008mg / kg, 0.01 mg / kg, 0.012 mg / kg, 0.015 mg / kg, 0.016mg / kg, 0.02 mg / kg, 0.021 mg / kg, 0.022mg / kg,0.023 mg / kg, or 0.025 mg / kg. For exemplary purposes, the dose comprises an mRNAconcentration of 0.33 μg / μL. For example, without being bound by any particular theory or methodology, the dose size is 500 μL, composed of 330 μL of mRNA (concentration of 500 μg / mL, i.e., 165 μg) and 170 μL of ethanol lipid nanoparticle solution. The ethanol may be removed after the particle formation via dialysis leaving the lipid nanoparticles containing mRNA in saline buffer alone. This may then be stored at 4oC until use. Thus, in some embodiments, the administered dose may comprise an mRNA concentration of 0.025 μg / μL – 0.33 μg / μL for the suspension of LNPs in saline buffer (preferably phosphate buffered saline)or any intermediate value encompassed therein, particularly, about: 0.025 μg / μL, 0.03 μg / μL,0.033 μg / μL, 0.035 μg / μL, 0.04 μg / μL, 0.045 μg / μL, 0.05 μg / μL, 0.055 μg / μL, 0.06 μg / μL,0.065 μg / μL 0.07 μg / μL, 0.075 μg / μL, 0.08 μg / μL, 0.085 μg / μL, 0.09 μg / μL, 0.095, μg / μL,0.1 μg / μL, 0.15 μg / μL, 0.2 μg / μL, 0.25 μg / μL, or 0.3 μg / μL, 0.33 μg / μL.In other aspects, provided herein is a composition containing the cells comprising the LNPs (e.g., the SLNs) disclosed herein, carrying the nucleic acids encoding the tumor protein p53 (e.g., p53 polypeptides) contemplated herein. The person of skill in the relevant art will appreciate that such cells may be administered via the adoptive transfer of said cells to a recipient subject in need thereof, as is known in the art. Briefly, and without being limited by theory, cells (selected from a third-party donor or cells derived from the recipient subject)may be brought into contact with the LNPs provided herein (e.g., in vitro or ex vivo), andadministered to the subject in need, by means known in the art. Therapeutic Methods In certain embodiments, provided herein are methods of treating a subject, comprising administering to the subject a therapeutic composition provided herein. Attorney Docket No.7504-00302 (GRH-00562) In some embodiments, the methods provided herein are used to treat or prevent Acute Lymphoblastic Leukemia (ALL), Acute Myeloid Leukemia (AML), AdrenocorticalCarcinoma, AIDS-Related Cancers (Kaposi Sarcoma (Soft Tissue Sarcoma), AIDS-RelatedLymphoma (Lymphoma), Primary CNS Lymphoma (Lymphoma)), Anal Cancer, AppendixCancer, Astrocytomas, Atypical Teratoid / Rhabdoid Tumor, Basal Cell Carcinoma of the Skin,Bile Duct Cancer, Bladder Cancer, Bone Cancer (includes Ewing Sarcoma and Osteosarcomaand Malignant Fibrous Histiocytoma), Brain Tumors, Breast Cancer, Bronchial Tumors(Lung Cancer), Burkitt Lymphoma, Carcinoma of Unknown Primary, Brain Cancer,Medulloblastoma and Other CNS Embryonal Tumors, Gemi Cell Tumor, Primary CNSLymphoma, Cervical Cancer, Childhood Cancers, Childhood Cardiac Tumors, Rarechildhood cancers (Nasopharyngeal Cancer, Esthesioneuroblastoma, Thyroid Cancer, OralCavity Cancer, Salivary Gland Tumors, Laryngeal Cancer and Papillomatosis, Midline TractCancer with NUT Gene Changes (NUT Midline Carcinoma)), Cholangiocarcinoma,Chordoma, Chronic Lymphocytic Leukemia (CLL), Chronic Myelogenous Leukemia (CML),Chronic Myeloproliferative Neoplasms, Colorectal Cancer, Craniopharyngioma, CutaneousT-Cell Lymphoma, Ductal Carcinoma In Situ (DCIS), Diffuse Intrinsic Pontine Glioma(DIPG), Endometrial Cancer, Ependymoma, Esophageal Cancer, Esthesioneuroblastoma,Ewing Sarcoma, Eye Cancer (including Intraocular Melanoma and Retinoblastoma),Fallopian Tube Cancer, Gallbladder Cancer, Gastric Cancer, Gastrointestinal Stromal Tumors(GIST), Germ Cell Tumors (including Extracranial Germ Cell Tumors, Extragonadal GermCell Tumors, Ovarian Germ Cell Tumors, and Testicular Cancer), Gestational TrophoblasticDisease, Hairy Cell Leukemia, Head and Neck Cancer, Heart Tumors, Hepatocellular Cancer,Langerhans Cell Histiocytosis, Hodgkin Lymphoma, Hypopharyngeal Cancer, Islet CellTumors, Pancreatic Neuroendocrine Tumors, Kidney or Renal Cell Cancer, LaryngealCancer, Leukemia, Lip and Oral Cavity Cancer, Liver Cancer, Lung Cancer (including (Non-Small Cell, Small Cell, Pleuropulmonary Blastoma, Pulmonary InflammatoryMyofibroblastic Tumors, and Tracheobronchial Tumors), Lymphoma, Male Breast Cancer,Melanoma, Merkel Cell Carcinoma, Mesothelioma, Metastatic Cancer, Metastatic SquamousNeck Cancer with Occult Primary, Mouth Cancer, Multiple Endocrine Neoplasia Syndromes,Multiple Myeloma / Plasma Cell Neoplasms, Mycosis Fungoides, MyelodysplasticSyndromes, Nasal Cavity and Paranasal Sinus Cancer, Nasopharyngeal Cancer, Neuroblastoma, Neuroendocrine Tumors, Non-Hodgkin Lymphoma, Oral Cancer, Oropharyngeal Cancer, Osteosarcoma and Undifferentiated Pleomorphic Sarcoma of Bone, Ovarian Cancer, Pancreatic Cancer, Papillomatosis, Paraganglioma, Paranasal Sinus and Attorney Docket No.7504-00302 (GRH-00562)Nasal Cavity Cancer, Parathyroid Cancer, Pheochromocytoma, Pituitary Tumor, Plasma CellNeoplasm (including Multiple Myeloma), Pleuropulmonary Blastoma, Pregnancy and Breast Cancer, Primary Central Nervous System (CNS) Lymphoma, Primary Peritoneal Cancer, Prostate Cancer, Pulmonary Inflammatory Myofibroblastic Tumors, Rectal Cancer, Recurrent Cancer, Renal Cell Cancer, Rhabdomyosarcoma, Salivary Gland Cancer, Various Sarcoma (including Uterine Sarcoma, Soft Tissue Sarcoma, and more), Sézary Syndrome, SkinCancer, Small Intestine Cancer, Squamous Cell Carcinoma of the Skin, Squamous NeckCancer with Occult Primary, Stomach (Gastric) Cancer, T-Cell Lymphoma, Throat Cancer(including Nasopharyngeal Cancer, Oropharyngeal Cancer, and Hypopharyngeal Cancer),Thymoma and Thymic Carcinoma, Thyroid Cancer, Tracheobronchial Tumors, TransitionalCell Cancer of the Renal Pelvis and Ureter, Urethral Cancer, Vaginal Cancer, VascularTumors, Vulvar Cancer, Wilms Tumor and Other Childhood Kidney Tumors, and otherdiseases and disorders resulting in tumors or cancer formations. Actual dosage levels of theactive ingredients in the pharmaceutical compositions provided herein may be varied so as toobtain an amount of the active ingredient which is effective to achieve the desired therapeuticresponse for a particular patient, composition, and mode of administration, without beingtoxic to the patient. The selected dosage level will depend upon a variety of factors including the activity of the particular agent employed, the route of administration, the time of administration, the rate of excretion or metabolism of the particular compound being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health, and prior medical history of the patient being treated, and like factors well known in the medical arts. In some embodiments, the methods provided herein further comprise treating the identified subject using a therapeutic method provided herein (e.g., by administering to the subject a composition provided herein). The administration of the disclosed compositions may be carried out in any convenient manner, including by injection, transfusion, or implantation, the compositions described herein may be administered to a patient subcutaneously, intradermally,intratumorally, intranodally, intramedullary, intramuscularly, by intravenous (i.v.) injection orinfusion, or intraperitoneally. In some embodiments, the disclosed compositions areadministered to a patient by intradermal or subcutaneous injection. In some embodiments, thedisclosed compositions are administered by i.v. injection or infusion. The compositions mayalso be injected directly into a tumor, lymph node, organ, or site of disease or disorder. In Attorney Docket No.7504-00302 (GRH-00562) certain embodiments, the disclosed compositions are administered to a patient in conjunction with (e.g., before, simultaneously or following) any number of relevant treatment modalities, including but not limited to standard-of-care treatment for a disease or condition contemplated herein, e.g., a cancer and / or tumor. In some embodiments, the compositions may be used in combination with chemotherapy, radiation, immunosuppressive agents, such as cyclosporine, azathioprine, methotrexate, mycophenolate, and FK506, antibodies, or other immunoablative agents such as CAM PATH, anti-CD3 antibodies or other antibody therapies, cytoxin, fludarabine, cyclosporin, FK506, rapamycin, mycophenolic acid, steroids, FR901228, cytokines, andirradiation. In some embodiments, the said compositions are administered to a patient inconjunction with (e.g., before simultaneously or following) bone marrow transplantation, T cell ablative therapy using either chemotherapy agents such as, fludarabine, external-beam radiation therapy (XRT), cyclophosphamide, or antibodies such as OKT3 or CAMPATH. Inother embodiments, the compositions disclosed herein are administered following B-cellablative therapy such as agents that react with CD20, e.g., Rituxan. For example, in someembodiments, subjects may undergo standard treatment with high-dose chemotherapy followed by peripheral blood stem cell transplantation. In certain embodiments, following the transplant, subjects receive an infusion of the cells disclosed herein. In additional embodiments, expanded cells are administered before or following surgery. EXAMPLES Example 1: Composition of Lipid Nanoparticle The composition contemplated herein may comprise four primary parts, an ionizable lipid, a phospholipid, a sterol, and a PEG-modified lipid. The LNPs are prepared by mixing an ethanolic lipid mixture with an acidic aqueous buffer containing the oligonucleotides of interest. A 1:3 ratio of ethanolic lipid mixture to aqueous buffer is generally used. For Example, the compositions comprises, 1. the ionizable lipid SM-1022. the phospholipid DSPC (distearoylphosphatidylcholine),3. cholesterol, and4. DMG-PEG 2000.Optionally, a secondary ionizable lipid may be used, such as ALC-0519 or ALC-0315, which has a PEG-lipid conjugate. A dose size of 500 μL was composed of 330 μL of mRNA and 170 μL of ethanol / lipid nanoparticle solution. For example, an initial mRNA payload concentration of 500 μg / mL Attorney Docket No.7504-00302 (GRH-00562) was diluted to achieve a desired mass ratio. In some embodiments, the mRNA to lipid mass- to-mass ratio is about 1:10. In some embodiments, the ethanol-to-water ratio may be about 1:3. The ethanol may be removed after the particle formation via dialysis leaving the lipid nanoparticles containing mRNA in PBS (phosphate buffered saline). This can then be run through a 220 nm filter to remove any aggregation that may have occurred during the dialysis process. Ethanolic Lipid Mixtures The four primary components comprises a molar ratio of about: 50 (SM-102):10 (DSPC):38.5 (Cholesterol):1.5 (PEG-Lipid)The molar ratio may be adjusted to optimally deliver the payload based on this starting ratio. For example, the ionizable lipid range can be adjusted ± 5, the DSPC can be adjusted ± 3, the cholesterol can be adjusted ± 5, and the DMG-PEG 2000 can be adjusted ± 0.5. Individual lipid stock solutions for each of the lipids (i.e., SM-102, DSPC, Cholesterol, and PEG-Lipid) in absolute ethanol were brought to room temperature prior to use, and the lipid mixture prepared as described in Table 2, which yielded 5 syntheses at a total volume of 4.0 mL. Amounts were calculated such that the ratio was kept at 50:10:38.5:1.5 molar ratio for ionizable lipid, DSPC, cholesterol, and PEG-lipid. Ethanol was used to dilute to the final volume. For example, 10.0 mg or DSPC was mixed with 400 μL ethanol, and so on, in accordance with Table 2 below. The appropriate volume of each lipid mixture component, as listed in Table 2, was then transferred to a single tube to prepare the ethanolic lipid mixture and pipetted several times to ensure mixing and avoid precipitation and cloudiness. Table 2 Component MolarMolar Concentration of mg needed Volumes each Volumes each, Mass Ratio stock in ethanol in 5 mL diluted to 5 mL diluted to 5 mL for 1.5 in total solution in total solution mg / mL for 1.5 g / L for 2.0 g / L SM-102 710.2 50 50 mg in 500 μL 4.27 mg 43 μL 57 μL(100 mg / mL) DSPC 790.145 10 10 mg in 400 μL at 0.95 mg 38 μL 51 μL35 mg / mL Cholesterol 386.65 38.5 5.0 mg in 1000 μL 1.79 mg 358 μL 477 μLat 5 mg / mL PEG-lipid 2509.2 1.5 5.0 mg in 1000 μL 0.45 mg 90 μL 120 μL Attorney Docket No.7504-00302 (GRH-00562) at 5 mg / mL Total Mass Lipids 7.46 mg in 5 mL9.95 mg in 5 mL lipid mixture lipid mixture stock solution stock solution Aqueous mRNA Solution: A lipid:mRNA (w:w) ratio of 10:1 and an ethanol aqueous ratio of 1:3 was used. Utilizing 0.50 mL from the 5.0 mL lipid mixture stock solution (e.g., for 1.5 g / L above), 75 μg mRNA was added to a separate tube and adjusted to a volume of 1.5 mL with 50 mM sodium acetate, pH 5.0 (e.g., 7.46 mg / 10= 0.75 mg; for a ratio of 10:1, 0.75 mg / 10 = 0.075 mg = 75 μg). The ethanolic lipid mixture was mixed with mRNA in a microfluidic mixer. The output, comprising the LNPs, were collected and injected into a dialysis cartridge. The cartridge was dialyzed in fresh PBS buffer to remove ethanol and loose lipids. The buffer was exchanged three times and the final LNP product was extracted and placed into a vessel for storage. Example 2: Payload An intended payload the LNPs was an mRNA strand which codes for the protein p53 which is located on the gene TP53. Utilizing specifically designed primers the following sequence was amplified and applied to the lipid nanoparticles disclosed herein. Table 3p53 Sequence FeaturesGTCTAGAGCCACCGTCCAGG (SEQ ID NO. 3) Forward Primers:TAATACGACTCACTATAGGTCTAGAGCCACCGTCCAGG (SEQ ID NO. 4) • T7 RNA polymerasePromoterGCAGGCCAACTTGTTCAGTG (SEQ ID NO. 5) Reverse Primer1 ctcaaaagtc tagagccacc gtccagggag caggtagctg ctgggctccg gggacactttp53 nucleotide sequence61 gcgttcgggc tgggagcgtg ctttccacga cggtgacacg cttccctgga ttggcagccadepicting complementary121 gactgccttc cgggtcactg ccatggagga gccgcagtca gatcctagcg tcgagcccccprimer sites:181 tctgagtcag gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtc241 ccccttgccg tcccaagcaa tggatgattt gatgctgtcc ccggacgata ttgaacaatg • forward primer301 gttcactgaa gacccaggtc cagatgaagc tcccagaatg ccagaggctg ctccccccgt • reverse primer361 ggcccctgca ccagcagctc ctacaccggc ggcccctgca ccagccccct cotggcccct • coding region421 gtcatcttct gtcccttccc agaaaaccta ccagggcagc tacggtttcc gtctgggctt481 cttgcattct gggacagcca agtctgtgac ttgcacgtac tcccctgccc tcaacaagat541 gttttgccaa ctggccaaga cctgccctgt gcagctgtgg gttgattcca cacccccgcc601 cggcacccgc gtccgcgcca tggccatcta caagcagtca cagcacatga cggaggttgt661 gaggcgctgc ccccaccatg agcgctgctc agatagcgat ggtctggccc ctcctcagca721 tcttatccga gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa acacttttcg781 acatagtgtg gtggtgccct atgagccgcc tgaggttggc tctgactgta ccaccatcca841 ctacaactac atgtgtaaca gttcctgcat gggcggcatg aaccggaggc ccatcctcac901 catcatcaca ctggaagact ccagtggtaa tctactggga cggaacagot ttgaggtgcg Attorney Docket No.7504-00302 (GRH-00562)961 tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa gagaatctcc gcaagaaagg1021 ggagcctcac cacgagctgc ccccagggag cactaagcga gcactgccca acaacaccag 1081 ctcctctccc cagccaaaga agaaaccact ggatggagaa tatttcaccc ttcagatccg 1141 tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag gccttggaac tcaaggatgc 1201 ccaggctggg aaggagccag gggggagcag ggctcactcc agccacctga agtccaaaaa 1261 gggtcagtct acctcccgcc ataaaaaact catgttcaag acagaagggc ctgactcaga1321 ctgacattct ccacttcttg ttccccactg acagcctccc acccccatct ctccctcccc1381 tgccattttg ggttttgggt ctttgaaccc ttgcttgcaa taggtgtgcg tcagaagcac 1441 ccaggacttc catttgcttt gtcccggggc tccactgaac aagttggcct gcactggtgt 1501 tttgttgtgg ggaggaggat ggggagtagg acataccagc ttagatttta aggtttttac 1561 tgtgagggat gtttgggaga tgtaagaaat gttcttgcag ttaagggtta gtttacaatc 1621 agocacatto taggtagggg cccacttcac cgtactaacc agggaagctg tccctcactg 1681 ttgaattttc tctaacttca aggcccatat ctgtgaaatg ctggcatttg cacctacctc 1741 acagagtgca ttgtgagggt taatgaaata atgtacatct ggccttgaaa ccacctttta 1801 ttacatgggg tctagaactt gacccccttg agggtgcttg ttccctctcc ctgttggtcg 1861 gtgggttggt agtttctaca gttgggcagc tggttaggta gagggagttg tcaagtctct 1921 gctggcccag ccaaaccctg tctgacaacc tcttggtgaa ccttagtacc taaaaggaaa 1981 tctcacccca tcccacaccc tggaggattt catctcttgt atatgatgat ctggatccac 2041 caagacttgt tttatgctca gggtcaattt cttttttctt tttttttttt ttttttcttt 2101 ttctttgaga ctgggtctcg ctttgttgcc caggctggag tggagtggcg tgatcttggc 2161 ttactgcagc ctttgcctcc ccggctcgag cagtcctgcc tcagcctccg gagtagctgg 2221 gaccacaggt tcatgccacc atggccagcc aacttttgca tgttttgtag agatggggtc 2281 tcacagtgtt gcccaggctg gtctcaaact cctgggctca ggcgatccac ctgtctcagc 2341 ctcccagagt gctgggatta caattgtgag ccaccacgtc cagctggaag ggtcaacatc 2401 ttttacattc tgcaagcaca tctgcatttt caccccaccc ttcccctcct tctccctttt 2461 tatatcccat ttttatatcg atctcttatt ttacaataaa actttgctgc ca (SEQ ID NO.2) guc uagagccacc guccagggag cagguagcug cugggcuccg gggacacuuu gcuucgggc Amplified mRNA sequenceugggagcgug cuuuccacga cggugacacg cuucccugga uuggcagcca gacugccuuccgggucacug ccauggagga gccgcaguca gauccuagcg ucgagccccc ucugagucag• coding regiongaaacauuuu cagaccuaug gaaacuacuu ccugaaaaca acguucuguc ccccuugccgucccaagcaa uggaugauuu gaugcugucc ccggacgaua uugaacaaug guucacugaagacccagguc cagaugaagc ucccagaaug ccagaggcug cuccccccgu ggccccugca ccagcagcuc cuacaccggc ggccccugca ccagcccccu ccuggccccu gucaucuucu gucccuuccc agaaaaccua ccagggcagc uacgguuucc gucugggcuu cuugcauucu gggacagcca agucugugac uugcacguac uccccugccc ucaacaagau guuuugccaa cuggccaaga ccugcccugu gcagcugugg guugauucca cacccccgcc cggcacccgc guccgcgcca uggccaucua caagcaguca cagcacauga cggagguugu gaggcgcugc ccccaccaug agcgcugcuc agauagcgau ggucuggccc cuccucagca ucuuauccga guggaaggaa auuugcgugu ggaguauuug gaugacagaa acacuuuucg acauagugug guggugcccu augagccgcc ugagguuggc ucugacugua ccaccaucca cuacaacuac auguguaaca guuccugcau gggcggcaug aaccggaggc ccauccucac caucaucaca cuggaagacu ccagugguaa ucuacuggga cggaacagcu uugaggugcg uguuugugcc uguccuggga gagaccggcg cacagaggaa gagaaucucc gcaagaaagg ggagccucac cacgagcugc ccccagggag cacuaagcga gcacugccca acaacaccag cuccucuccc cagccaaaga agaaaccacu ggauggagaa uauuucaccc uucagauccg ugggcgugag cgcuucgaga uguuccgaga gcugaaugag gccuuggaac ucaaggaugc ccaggcuggg aaggagccag gggggagcag ggcucacucc agccaccuga aguccaaaaa gggucagucu accucccgcc auaaaaaacu cauguucaag acagaagggc cugacucaga cugacauucu ccacuucuug uuccccacug acagccuccc acccccaucu cucccucccc ugccauuuug gguuuugggu cuuugaaccc uugcuugcaa uaggugugcg ucagaagcac ccaggacucc cauuugcuuu gucccggggc uccacugaac aaguuggccu gc (SEQ ID NO. 6) The amplified mRNA sequence was inserted into the lipid nanoparticles (LNPs) of the invention. Such mRNA sequences comprised modifications to the 5’ cap, and optionally substitution some or all of the uridines with pseudouridines, as disclosed herein. Example 3: Payload II Attorney Docket No.7504-00302 (GRH-00562) An intended payload for the LNPs was an mRNA strand which codes for the protein p53 which is located on the gene TP53. Utilizing specifically designed primers the following sequence was amplified and applied to the lipid nanoparticles disclosed herein. Table 4p53 Sequence FeaturesGTGCTTTCCACGACGGTGA (SEQ ID NO. 7)Forward Primers:TAATACGACTCACTATAG GTGCTTTCCACGACGGTGA (SEQ ID NO. 8) • T7 RNA polymerasePromoterAAACTACCAACCCACCGACC (SEQ ID NO. 9) Reverse Primer1 ctcaaaagtc tagagccacc gtccagggag caggtagctg ctgggctccg gggacactttp53 nucleotide sequence61 gcgttcgggc tgggagcgtg ctttccacga cggtgacacg cttccctgga ttggcagccadepicting complementary121 gactgccttc cgggtcactg ccatggagga gccgcagtca gatcctagcg tcgagcccccprimer sites:181 tctgagtcag gaaacatttt cagacctatg gaaactactt cctgaaaaca acgttctgtc241 ccccttgccg tcccaagcaa tggatgattt gatgctgtcc ccggacgata ttgaacaatg • forward primer301 gttcactgaa gacccaggtc cagatgaagc tcccagaatg ccagaggctg ctccccccgt • reverse primer361 ggcccctgca ccagcagctc ctacaccggc ggcccctgca ccagccccct cotggcccct • coding region421 gtcatcttct gtcccttccc agaaaaccta ccagggcagc tacggtttcc gtctgggctt481 cttgcattct gggacagcca agtctgtgac ttgcacgtac tcccctgccc tcaacaagat541 gttttgccaa ctggccaaga cctgccctgt gcagctgtgg gttgattcca cacccccgcc601 cggcacccgc gtccgcgcca tggccatcta caagcagtca cagcacatga cggaggttgt661 gaggcgctgc ccccaccatg agcgctgctc agatagcgat ggtctggccc ctcctcagca721 tcttatccga gtggaaggaa atttgcgtgt ggagtatttg gatgacagaa acacttttcg781 acatagtgtg gtggtgccct atgagccgcc tgaggttggc tctgactgta ccaccatcca841 ctacaactac atgtgtaaca gttcctgcat gggcggcatg aaccggaggc ccatcctcac901 catcatcaca ctggaagact ccagtggtaa tctactggga cggaacagot ttgaggtgcg961 tgtttgtgcc tgtcctggga gagaccggcg cacagaggaa gagaatctcc gcaagaaagg1021 ggagcctcac cacgagctgc ccccagggag cactaagcga gcactgccca acaacaccag 1081 ctcctctccc cagccaaaga agaaaccact ggatggagaa tatttcaccc ttcagatccg 1141 tgggcgtgag cgcttcgaga tgttccgaga gctgaatgag gccttggaac tcaaggatgc 1201 ccaggctggg aaggagccag gggggagcag ggctcactcc agccacctga agtccaaaaa 1261 gggtcagtct acctcccgcc ataaaaaact catgttcaag acagaagggc ctgactcaga1321 ctgacattct ccacttcttg ttccccactg acagcctccc acccccatct ctccctcccc1381 tgccattttg ggttttgggt ctttgaaccc ttgcttgcaa taggtgtgcg tcagaagcac 1441 ccaggacttc catttgcttt gtcccggggc tccactgaac aagttggcct gcactggtgt 1501 tttgttgtgg ggaggaggat ggggagtagg acataccagc ttagatttta aggtttttac 1561 tgtgagggat gtttgggaga tgtaagaaat gttcttgcag ttaagggtta gtttacaatc 1621 agocacatto taggtagggg cccacttcac cgtactaacc agggaagctg tccctcactg 1681 ttgaattttc tctaacttca aggcccatat ctgtgaaatg ctggcatttg cacctacctc 1741 acagagtgca ttgtgagggt taatgaaata atgtacatct ggccttgaaa ccacctttta 1801 ttacatgggg tctagaactt gacccccttg agggtgcttg ttccctctcc ctgttggtcg1861 gtgggttggt agtttctaca gttgggcagc tggttaggta gagggagttg tcaagtctct1921 gctggcccag ccaaaccctg tctgacaacc tcttggtgaa ccttagtacc taaaaggaaa1981 tctcacccca tcccacaccc tggaggattt catctcttgt atatgatgat ctggatccac 2041 caagacttgt tttatgctca gggtcaattt cttttttctt tttttttttt ttttttcttt 2101 ttctttgaga ctgggtctcg ctttgttgcc caggctggag tggagtggcg tgatcttggc 2161 ttactgcagc ctttgcctcc ccggctcgag cagtcctgcc tcagcctccg gagtagctgg 2221 gaccacaggt tcatgccacc atggccagcc aacttttgca tgttttgtag agatggggtc 2281 tcacagtgtt gcccaggctg gtctcaaact cctgggctca ggcgatccac ctgtctcagc 2341 ctcccagagt gctgggatta caattgtgag ccaccacgtc cagctggaag ggtcaacatc 2401 ttttacattc tgcaagcaca tctgcatttt caccccaccc ttcccctcct tctccctttt 2461 tatatcccat ttttatatcg atctcttatt ttacaataaa actttgctgc ca (SEQ ID NO.2)gug cuuuccacga cggugacacg cuucccugga uuggcagcca gacugccuuc cgggucacugAmplified mRNA sequenceccauggagga gccgcaguca gauccuagcg ucgagccccc ucugagucag gaaacauuuucagaccuaug gaaacuacuu ccugaaaaca acguucuguc ccccuugccg ucccaagcaa • coding regionuggaugauuu gaugcugucc ccggacgaua uugaacaaug guucacugaa gacccagguc Attorney Docket No.7504-00302 (GRH-00562)cagaugaagc ucccagaaug ccagaggcug cuccccccgu ggccccugca ccagcagcuccuacaccggc ggccccugca ccagcccccu couggccccu gucaucuucu gucccuucccagaaaaccua ccagggcagc uacgguuucc gucugggcuu cuugcauucu gggacagccaagucugugac uugcacguac uccccugccc ucaacaagau guuuugccaa cuggccaagaccugcccugu gcagcugugg guugauucca cacccccgcc cggcacccgc guccgcgccauggccaucua caagcaguca cagcacauga cggagguugu gaggcgcugc ccccaccaugagcgcugcuc agauagcgau ggucuggccc cuccucagca ucuuauccga guggaaggaaauuugcgugu ggaguauuug gaugacagaa acacuuuucg acauagugug guggugcccuaugagccgcc ugagguuggc ucugacugua ccaccaucca cuacaacuac auguguaacaguuccugcau gggcggcaug aaccggaggc ccauccucac caucaucaca cuggaagacuccagugguaa ucuacuggga cggaacagou uugaggugcg uguuugugcc uguccugggagagaccggcg cacagaggaa gagaaucucc gcaagaaagg ggagccucac cacgagcugcccccagggag cacuaagcga gcacugccca acaacaccag cuccucuccc cagccaaagaagaaaccacu ggauggagaa uauuucaccc uucagauccg ugggcgugag cgcuucgagauguuccgaga gcugaaugag gccuuggaac ucaaggaugc ccaggcuggg aaggagccaggggggagcag ggcucacucc agccaccuga aguccaaaaa gggucagucu accucccgccauaaaaaacu cauguucaag acagaagggc cugacucaga cugacauucu ccacuucuuguuccccacug acagccuccc acccccaucu cucccucccc ugccauuuug gguuuugggucuuugaaccc uugcuugcaa uaggugugcg ucagaagcac ccaggacuuc cauuugcuuugucccggggc uccacugaac aaguuggccu gcacuggugu uuuguugugg ggaggaggauggggaguagg acauaccagc uuagauuuua agguuuuuac ugugagggau guuugggagauguaagaaau guucuugcag uuaaggguua guuuacaauc agocacauuo uagguaggggcccacuucac cguacuaacc agggaagcug ucccucacug uugaauuuuc ucuaacuucaaggcccauau cugugaaaug cuggcauuug caccuaccuc acagagugca uugugaggguuaaugaaaua auguacaucu ggccuugaaa ccaccuuuua uuacaugggg ucuagaacuugacccccuug agggugcuug uucccucucc cuguuggucg guggguuggu aguuu(SEQ ID NO. 10) The amplified mRNA sequence was inserted into the lipid nanoparticles (LNPs) of the invention. Such mRNA sequences comprised modifications to the 5’ cap, and optionally substituting some or all of the uridines with pseudouridines, as disclosed herein. Example 4: Dynamic Light Scattering (DLS) In order for the LNP formulations disclosed herein to be used as a cancer treatment,each particle must be small enough to pass through the blood brain barrier (to treat any formsof intracranial tumors). Particles that are less than 200 nm (and preferably about 100 nm) areable to efficiently pass through the blood brain barrier (Ceña & Jávita, 2018). Doses of LNP-encapsulated mRNAs were tested with a Dynamic Light Scattering (DLS) machine. In DLS, when laser light encounters macromolecules in a solution the incident light scatters in all directions and scattering intensity is recorded by a detector. The rate of fluctuations in scattered light is directly related to the rate of diffusion of the particle through the solvent,which is related in turn to the particles' hydrodynamic radii. Smaller particles diffuse faster,causing more rapid fluctuations in the intensity than larger particles. Therefore, thefluctuation in light intensity contains information about the diffusion of the molecules andcan be used to extract a diffusion coefficient and calculate a particle size. Continuous DLS data collection showed that the LNP formulations disclosed herein consistently comprised particles with a diameter of 110 nm or smaller. In a typical DLS Attorney Docket No.7504-00302 (GRH-00562)analysis the Z-Average is the estimated average size of the particles being measured. The D50showed that 50% of the particles are the reported size or below. For example, Figure 3 showsthat 50% of the particles are estimated to be 90.9 nm or below in size (see HistogramOperations: % Cumulative (6), the aforementioned D50). The Count Rate (measured in kilocounts per second (kCPS)) correlates with the concentration of the sample being measured. ACount Rate above 400 kCPS is acceptable for the particles of the invention disclosed herein.Higher Count Rates can indicate a more concentrated dose, i.e., that there are more particlescontaining the p53 mRNA. This without being bound by theory or methodology the higherthe concentration, the more effective that dose can be.LNP solutions were filter sterilized with a 0.22 μm filter and stored at 4°C until use. Optionally, the LNPs can be lyophilized and stored at -80°C for long-term storage. DLS tests were run at least in triplicate. Thus, at least three measurements were taken of a sample, as illustrated in the overlay of the Z-Average (measure of the average size of a particle size distribution) depicted in Figure 4. Notably, there is little variance among these measurements resulting in a single visible peak. (86.9 nm to 88.7 nm, (all three replicates laid out over each other). This data does not reflect the average kCPS of all three measurements as this outputwas used to compare triplicate measurements to each other.In addition to testing each batch of doses made, doses were saved to measure the size change over time. Such doses were used to measure if the LNPs encapsulating mRNA were aggregating. Even minor aggregation would be detectable. Generally, for DLS measure in intensity, one larger particle can block out many more smaller particles and skew the data to show a larger Z-Average and D50 than is actually true. The test sample was prepared andstored at 4°C (the temperature at which all tested doses are stored). Particle size wasmeasured at several different time points following completion of initial dialysis andfiltration. To re-measure, at each time point a portion of sample was applied to a cuvette andread on the DLS machine and retired, e.g., to a 15 mL conical tube for storage at 4°C again.Results showed that even after 145 days in storage, there was no aggregation of particles.Thus, the LNPs of the invention are stable at 4°C for long periods of time without changingsize. In summary, the DLS data showed that particles at 110 nm or smaller can be made consistently. This will allow LNPs to cross the blood-brain barrier to deliver p53 mRNA tothe cells of the brain. Said particles do not aggregate together or change in size over time, andcan be stored at 4°C with no degradation or aggregation over 145 days of repeated testing. Attorney Docket No.7504-00302 (GRH-00562) Example 5 synthesis PCR (DNA synthesis): Plasmids containing the nucleotide sequence for p53 were used in the synthesis and amplification of double stranded DNA containing the nucleotide sequence of the p53 protein.Commercially available kits, such as the LongAmp® Taq PCR Kit, were used. In a singlereaction mixture, the enzyme LongAmp® Taq DNA polymerase was used to create DNAfrom a p53 plasmid template with the forward and reverse primer pairs disclosed herein (e.g., SEQ ID NOs. 4 and 5, 8 and 9, 12 and 13, and 16 and 17). This double stranded DNA is coded for the TP53 gene. Synthesis reactions were run with the following thermal cycler parameters: Table 5 InitialDenature Anneal Extend FinalFinal Hold Denaturation Extension 1cycle 30 cycles 1 cycle 1 cycle94°C 94°C 55°C 65°C 65°C 4°C30 seconds 20 seconds 35 seconds 1 minute and 10 minutes indefinite25 seconds After the cycler finished running, the reaction tubes could be stored at -20°C freezer,otherwise the resultant double-stranded DNA was used immediately for mRNA creation. DNA Agarose Analysis: Samples were prepared for loading onto agarose gel by taking 1 μL of DNA sample with 3 μL of 1X loading dye (4 μL total), pipetted up and down slowly 3-5 times to mix. This was done with each sample run on an agarose gel. A1KB DNA ladder was applied to the first well and the 4 μL samples were added toseparate wells (i.e., up to 9 DNA samples in each agarose gel). Samples were electrophoresed for 1 hour at a constant 100V. The expected length of the sequence should be ~1500 base pairs which was observed for all 4 samples shown in the p53 DNA Agarose Gel of Figure 1. The nucleotide sequence of the double-stranded DNA was assessed and confirmed to be the intended target sequence by Sanger sequencing (i.e., chain termination method) analysis. mRNA synthesis: Attorney Docket No.7504-00302 (GRH-00562) The prepared DNA was used in the synthesis of p53 mRNA. Commercially available kits, such as the HiScribe® T7 Quick Yield RNA Synthesis Kit, were used. Thermal cycler reactions were run according to kit manufacture protocols. Once thermal cycling was complete, the mRNA was capped using a Fausto virus capping enzyme (FCE) and Cap-2’-O-methyltransferase. For each tube 24.2 μL of Nuclease- Free Water was added, followed by 6.5 μL of FCE capping buffer, 3.25 μL of S-Adenosyl methionine (SAM) (32 mM), 3.25 μL of Guanosine triphosphate (GTP) (10 mM), 2.6 μL of the FCE enzyme (25,000 units / mL), and 5.2 μL of Cap 2’-O-methyltransferase (50,000 units / mL) was added in that order, giving a final volume of 65 μL. The tubes were thenincubated in a thermal cycler and set for a 1-hour hold at 37°C followed by an indefinite 4°Chold. RNA purification and concentration following enzymatic reaction was performedusing commercially available kits such as the Monarch® RNA Clean up Kit. Briefly, 100 μLof RNA Binding Buffer was added to the 65 μL mRNA synthesis and mixed by pipetting. Thetotal 165 μL of solution was added to the spin column provided with 165 μL of 100% ethanol and carefully pipetted to mix. Spin columns (with collection tubes) were centrifuged for 1 minute at 13,000 rpm. Liquid flowthrough (liquid in collection tube) was removed and 500 μL of appropriate RNA wash buffer was added to the spin column. Following centrifugationfor 1 minute at 13,000 rpm, the liquid flow through was removed, and the wash repeated atleast once more.Following washing, 50-100 μL of nuclease free water was added to the spin column (with a new empty collection tube) and incubated at room temperature for 5 minutes. After the 5 minute incubation, the spin column (with empty collection tube) was centrifuged for 1 minute at 13,000 rpm. The collection tine containing the resultant flowthrough was held at4°C. The purified mRNA from multiple collection tubes were combined and mixed viapipetting. Following collection of purified RNA, the concentration of the combined RNA wasdetermined by using a Nanophotometer. Purified p53 RNA could be stored at 4°C until use.Optionally a DNase step can be performed prior to the capping step. This would be done by adding 30 μL of water to the mRNA tube followed by 2 μL of the DNase enzyme(2,000 units / mL). This would then be incubated at 37°C for 15 minutes. After this incubationfinishes, the mRNA would proceed to mRNA capping step. mRNA Agarose Gel Analysis: Samples were prepared for loading onto a 1% agarose gel by diluting 1 μL of mRNA to a concentration of about 300-400 μg / mL in nuclease free water. This diluted mRNA was Attorney Docket No.7504-00302 (GRH-00562) mixed by slowly pipetting up and down 3-5 times. 1 μL of diluted mRNA sample was added to 3 μL of 2X RNA loading dye, pipetting up and down slowly 3-5 times to mix. A ssRNA ladder was applied to the first well and the 4 μL sample(s) were added to each subsequent well (i.e., synthesized mRNA from a day followed by synthesized mRNA from another day). Samples were electrophoresed for 1 hour at a constant 100 V. Multiple separate p53 mRNA synthesis runs yielded identical results and produced mRNA for p53 at ~1500 bases long in high concentrations (see Figure 2). This mRNA represents the sequence (or payload) disclosed herein and incorporated into the LNPs of the invention In summary, p53 DNA was created from a p53 plasmid template. The DNApolymerase enzyme LongAmp® Taq DNA Polymerase was added to the p53 plasmidtemplate to create double stranded DNA coding for the p53 gene using the appropriateprimers. This 1485 base pair long strand was consistently synthesized as shown in Figure 1.The double-stranded DNA coding for the TP53 gene was then used to create mRNA, as theforward primer used to synthesize DNA contained a promoter for T7 RNA polymerase. Afterthe mRNA synthesis, a 5’ cap was added with a Faustovirus Capping Enzyme (FCE) and Cap2’-O-methyltransferase. After the cap was added, the resultant sequence was then cleaned toremove any impurities such as free-floating nucleotides, enzymes, DNA, etc. The fullycleaned and capped mRNA is shown in Figure 2 and represents Payload 1 used to create afinal lipid nanoparticle containing p53 mRNA product contemplated herein.Example 6: Cellular Viability Testing p53 is one of the main proteins involved with apoptosis in cells. It is also often mutated to become ineffective or is inhibited in many forms of cancer. To test the effectiveness of p53 mRNA-LNPs on the treatment of cancers, cellular viability testing was run on both SW-1417 and SK-N-FI human neuroblastoma cells. Experiments were performed 24 and 48 hours after the initial dosing of cells with p53 mRNA-LNPs or Empty LNPs. The p53 mRNA-LNP dosed SW-1417 cells saw a decrease in viability between 50% and 70% when compared to SW-1417 cells dosed with Empty LNPs. The p53 mRNA-LNP dosed SK- N-FI cells saw a decrease in viability between 11% and 21% when compared to SK-N-FI cells dosed with Empty LNPs. Both cell types also show a significant decrease in the total number of cells in each well when treated with p53 mRNA-LNPs instead of Empty LNPs. All experiments were performed in 12 well plates where each well was seeded with 250,000 cells and 1 mL of complete DMEM media. They were then allowed to grow and adhere overnight in an incubator set to 37oC / 5% CO2. Attorney Docket No.7504-00302 (GRH-00562) 24-hour cell count: After being allowed to grow and adhere overnight, 6 wells of a 12-LNPs. The other 6 wells were dosed with 40 oC / 5% CO2) for 24 hours. After the 24-hour incubation, the cell viability in each well was assessedby microscopy on a hemocytometer using trypan blue staining. See figures 5 and 6 for theview of SW-1417 cells dosed with empty LNPs and p530 mRNA-LNPs respectively. SeeFigures 7-10 for results of the assay. 48-hour (single dosed) cell count: After being allowed to grow and adhere overnight,6 wells of a 12 well plate (seeded with SW- mRNA- was then returned to a cell incubator (set to 37oC / 5% CO2) for 48 hours. After the 48-hour incubation, the cell viability in each well was assessed by microscopy on a hemocytometerusing trypan blue staining. See figures 11 and 12.48-hour (dosing every 24 hours) cell count: After being allowed to grow and adhere overnight, 6-LNPs. The other 6 wells were dosed oC / 5% CO2) for 24 hours. After this 24-hour incubation, the cells were dosed again following the same procedure as above. The 12 well plate was then returned to the incubator for another 24-hour incubation period. After the second 24-hour incubation, the cell viability in each well was assessed bymicroscopy on a hemocytometer using trypan blue staining. See figures 13-16.Example 7: Efficacy testing in PDX-NSG miceTo test p53 mRNA-LNPs’ ability to treat cancer, such as colorectal adenocarcinoma, 15 PDX-NSG mice were xenografted with colorectal adenocarcinoma onto their lower right side. After the tumors were seen to grow into measurable sizes on all the mice, 12 of the mice were dosed with p53 mRNA-LNPs. These mice were split into 6 different groups. Group Awas dosed with 75 of p53 mRNA-LNPs, group B was dosed with 15 of p53 mRNA-LNPs,group C was dosed with 8 of p53 mRNA-LNPs, group D was dosed with 4 of p53 mRNA-LNPs, group E was dosed with 2 of p53 mRNA-LNPs, and group F was dosed with 1 of p53mRNA-LNPs. 2 of the remaining 3 mice were dosed with 75 of PBS (group G). One mouse ofeach group was dosed with the respective treatment via a lateral tail vein intravenous injection. The other mouse of each group was dosed with the respective treatment via direct injection into the tumor.The 15th mouse was left undosed (group H). Dosing occurred every 48 hours for 21 days. Tumorsizes were measured every day and mouse weights were recorded everyday. After the 21st day of dosing, all mice were euthanized, and the tumors of each mouse were removed for tumor weight measurements. See Figure 17 for images of all removed tumors. Attorney Docket No.7504-00302 (GRH-00562) Table 6 Mouse Dose Volume and Route ofFinal Tumor Weight Tumor Volume Administration (g) Change from Day 1 (mm3)Group A #1 75 p53-mRNA-LNP / Direct 0.417 285.95InjectionGroup A #2 75 p53-mRNA-LNP / I.V. 0.116 275.49InjectionGroup B #1 15 p53-mRNA-LNP / Direct 0.240 29.37InjectionGroup B #2 15 p53-mRNA-LNP / I.V. 0.088 99.36InjectionGroup C #1 8 p53-mRNA-LNP / Direct 0.407 259.81InjectionGroup C #2 8 p53-mRNA-LNP / I.V. 0.380 327.01InjectionGroup D #1 4 p53-mRNA-LNP / Direct 0.415 194.13InjectionGroup D #2 4 p53-mRNA-LNP / I.V. 0.191 -14.01InjectionGroup E #1 2 p53-mRNA-LNP / Direct 0.346 188.42InjectionGroup E #2 2 p53-mRNA-LNP / I.V. 0.143 229.69InjectionGroup F #1 1 p53-mRNA-LNP / Direct 0.709 326.79InjectionGroup F #2 1 p53-mRNA-LNP / I.V. 0.358 735.65InjectionGroup G #1 75 PBS / Direct Injection 0.741 525.82Group G #2 75 PBS / I.V. Injection 0.293 463.52Group H #1 NA (mouse was undosed during 0.530 393.37trial) The average change in tumor volume of the control mice (groups G and H) was+494.24 mm3. The change in tumor volume of both mice in groups A, B, C, D, and E wassignificantly less than the average change in control mice tumor volume. In group F, the Attorney Docket No.7504-00302 (GRH-00562) change in tumor volume was either not significantly different, or greater than the averagecontrol. This suggested that the 1 p53-mRNA-LNP dose was ineffective in treating thexenografted tumor. Groups A, B, C, D, and E clearly display the ability of a p53 mRNA lipidnanoparticle to slow or halt the growth of colorectal adenocarcinoma as all tumors grewsignificantly slower than the control tumors with the 4 p53-mRNA-LNP dosed via I.V.injection tumor shrinking in size. Overall, the results of this study suggest that p53 mRNA-LNPs are effective in slowing the growth of colorectal adenocarcinoma tumors in mice.INCORPORATION BY REFERENCE All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present specification,including its specific definitions, will control. While specific aspects of the patient matterhave been discussed, the above specification is illustrative and not restrictive. Manyvariations will become apparent to those skilled in the art upon review of this specificationand the claims below. The full scope should be determined by reference to the claims, alongwith their full scope of equivalents, and the specification, along with such variations.EQUIVALENTS The present invention has been described in connection with what are presently considered to be the most practical and preferred embodiments. However, the invention hasbeen presented by way of illustration and is not intended to be limited to the disclosedembodiments. Accordingly, one of skill in the art will realize that the invention is intended toencompass all modification and alternative arrangements within the spirit and scope as setforth in the appended claims.

Claims

Attorney Docket No.7504-00302 (GRH-00562) What is claimed is:

1. A therapeutic composition comprising a nucleic acid formulated in a lipidnanoparticle (LNP), wherein the nucleic acid comprises an open reading frame encoding tumor protein p53 polypeptide, or a functional fragment thereof.

2. The therapeutic composition of claim 1, where in the tumor protein p53 polypeptide iscellular tumor antigen p53 isoform a transcript variant 1, cellular tumor antigen p53 isoform a transcript variant 2, cellular tumor antigen p53 isoform a transcript variant 9, cellular tumor antigen p53 isoform a transcript variant 10, cellular tumor antigen p53 isoform a transcript variant 11, cellular tumor antigen p53 isoform b transcript variant 3, cellular tumor antigen p53 isoform b transcript variant 12, cellular tumor antigen p53 isoform b transcript variant 13, cellular tumor antigen p53 isoform c, cellular tumor antigen p53 isoform d, cellular tumor antigen p53 isoform e, cellular tumor antigen p53 isoform f, cellular tumor antigen p53 isoform g transcript variant 8, cellular tumor antigen p53 isoform g transcript variant 1, cellular tumor antigen p53 isoform g transcript variant 2, cellular tumor antigen p53 isoform g transcript variant 9, cellular tumor antigen p53 isoform g transcript variant 10, cellular tumor antigen p53 isoform g transcript variant 11, cellular tumor antigen p53 isoform h, cellular tumor antigen p53 isoform i transcript variant 3, cellular tumor antigen p53 isoform i transcript variant 12, cellular tumor antigen p53 isoform i transcript variant 13, cellular tumor antigen p53 isoform, cellular tumor antigen p53 isoform k, or cellular tumor antigen p53 isoform 1.

3. The therapeutic composition of claim 1, wherein the open reading frame is derivedfrom the nucleic acid sequence set forth in SEQ ID NO. 2, or a functional fragment thereof.

4. The therapeutic composition of claim 1, wherein the nucleic acid is mRNA, optionallywherein the mRNA comprises the nucleic acid sequence set forth in any one of SEQ ID NO. 6, SEQ ID NO. 10, or any functional fragment thereof.

5. The therapeutic composition of any one of claims 1-4, wherein the LNP comprises anionizable lipid, a phospholipid, a sterol, a PEG-modified lipid, or any combination thereof.Attorney Docket No.7504-00302 (GRH-00562)6. The therapeutic composition of any one of claims 1-4, wherein the LNP consistsessentially of an ionizable lipid, a phospholipid, a sterol, a PEG-modified lipid, or any combination thereof.

7. The therapeutic composition of claim 5 or 6, wherein the ionizable lipid is SM-102.

8. The therapeutic composition of any one of claims 5-7, wherein the phospholipid isdistearoylphosphatidylcholine (DSPC).

9. The therapeutic composition of any one of claims 5-8, wherein the sterol ischolesterol.

10. The therapeutic composition of any one of claims 5-9, wherein the PEG-modifiedlipid is DMG-PEG 2000.

11. A cell comprising the LNP of any one of claims 1-10.

12. The cell of claim 11, expressing the p53 polypeptide encoded by an mRNA.

13. The cell of claim 12, wherein the p53 polypeptide comprises the amino acid sequenceset for in SEQ ID NO. 1, or a functional fragment thereof.

14. The cell of any one of claims 11-13, wherein the cell is an endothelial cell, epithelialcell, neuronal cell, non-neuronal cell, or haematopoietic cell.

15. The cell of claim 14, wherein the haematopoietic cell is an immune cell selected froma lymphocyte, a monocyte, a dendritic cell, a mast cell, a neutrophil, a basophil, or an eosinophil.

16. The cell of claim 15, wherein the immune cell is lymphocyte selected from a T cell,T cell, a Natural Killer (NK) cell, a Natural Killer T (NKT) cell, a B cell, an innate lymphoid cell (ILC), a cytokine induced killer (CIK) cell, a cytotoxic T lymphocyte (CTL), a lymphokine activated killer (LAK) cell, or a regulatory T cell.Attorney Docket No.7504-00302 (GRH-00562)17. The cell of any one of claims 11-16, wherein the cell is a cell derived from bonemarrow.

18. The cell of any one of claims 11-14, wherein the cell is a cell of the central nervoussystem (CNS) or peripheral nervous system (PNS).

19. The cell of any one of claims 11-18, wherein the cell is a cell present in the CNS.

20. The cell of 18 or 19, wherein the cell is a neuronal cell.

21. The cell of claim 20, wherein the nerve cell is a sensory neuron, a motor neuron, or aninterneuron.

22. The cell of claim 18 or 19, wherein the cell is a non-neuronal cell.

23. The cell of claim 22, wherein the non-neuronal cell is a glial cell.

24. The cell of claim 23, wherein the glial cell is an astrocyte cell, an oligodendrocytecell, an ependymal cell, a radial glial cell, a Schwann cell, a satellite cell, an enteric glial cell, or a microglial cell.

25. A method of treating cancer in a subject, the method comprising administering thetherapeutic composition of any one of claims 1-10.

26. A method of treating cancer in a subject, the method comprising administering acomposition comprising the cells of any one of claims 11-24.

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