HERPES VIRUS, USES THEREOF, AND, METHODS FOR INTENSIFYING, INCREASING, ELEVATING, AND / OR SUPPLEMENTING THE LEVELS OF A CFTR POLYPEPTIDE IN ONE OR MORE CELLS, TO REDUCE OR INHIBIT PROGRESSIVE DESTRUCTION OF THE LUNG, TO PROVIDE PROPHYLAXIAL, PALLIATIVE, OR THERAPEUTIC RELIEF OF ONE OR MORE SIGNS OR SYMPTOMS OF CYSTIC FIBROSIS AND TO PROVIDE PROphylactic, PALLIATIVE OR THERAPEUTIC RELIEF OF ONE OR MORE SIGNS OR SYMPTOMS OF COPD

Description

Herpes virus, its uses, and methods for intensifying, increasing, raising, and / or supplementing the levels of a CFTR polypeptide in one or more cells, to reduce or inhibit progressive lung destruction, to provide prophylactic, palliative, or therapeutic relief of one or more signs or symptoms of cystic fibrosis, and to provide prophylactic, palliative, or therapeutic relief of one or more signs or symptoms of COPD. Split from BR112021014400-5, filed on 07 / 02 / 2020. CROSS-REFERENCE TO RELATED ORDERS.

[001] This application claims priority benefit of the Application Provisional US 62 / 802,871, filed February 8, 2019, which is incorporated herein by reference in its entirety. PRESENTATION OF A SEQUENCE LISTING IN AN ASCII TEXT FILE

[002] The contents of the following submission in ASCII text file are incorporated here by reference in their entirety: a machine-readable form (CRF) of the Sequence Listing (filename: 7613420001140SEQLIST.txt, registration date: January 17, 2020, size: 44 KB). FIELD OF THE INVENTION

[003] This disclosure relates, in part, to recombinant nucleic acids comprising one or more polynucleotides encoding a cystic fibrosis transmembrane conductance regulator (CFTR) polypeptide, viruses comprising the same, pharmaceutical compositions and formulations thereof, and methods of their use (for example, to provide prophylactic, palliative or therapeutic relief of one or more signs or symptoms of a chronic lung disease such as cystic fibrosis). FOUNDATION Petition 870260061885, dated 06 / 24 / 2026, page 15 / 151 / 101

[004] The cystic fibrosis transmembrane conductance regulator (CFTR) is a cAMP-activated bicarbonate and chloride channel that is critical for pulmonary homeostasis. Reduced or lost CFTR channel function generally leads to mucus stasis, chronic bacterial infections, and the accompanying chronic inflammatory responses, which promote progressive lung destruction. It has been suggested that decreases in CFTR expression are a component of the pulmonary pathology observed in patients with chronic obstructive pulmonary disease (COPD), and loss-of-function mutations in the CFTR gene lead to the dire consequences associated with cystic fibrosis (CF). More than 2,000 unique mutations in the CFTR gene have been described.

[005] CF is a hereditary disease characterized by the accumulation of thick, sticky mucus that can damage many organs in the body; however, the most serious pathological consequences are associated with the lungs. Patients with CF have dehydrated mucus in the lungs leading to airway obstruction, chronic bacterial infections (and associated inflammatory responses), bronchiectasis, and ultimately respiratory failure. Currently, more than 70,000 people live with cystic fibrosis worldwide. Historically, children born with CF died as infants, and in 1980, the average survival was less than 20 years.Although medical advances over the past three decades have dramatically improved the quality of life and life expectancy of CF patients (40.6 years in the United States as of 2013), there is a clear need for new treatment options aimed at the molecular correction of CFTR deficiencies observed in CF patients, as well as in patients suffering from other chronic lung diseases such as COPD.

[006] All references cited herein, including patent applications, patent publications, non-patent literature and NCBI / UniProtKB / Swiss-Prot accession numbers, are incorporated herein by reference. Petition 870260061885, dated 06 / 24 / 2026, page 16 / 151 / 101 in its entirety, as if each individual reference were specifically and individually indicated to be incorporated by reference. BRIEF SUMMARY

[007] In order to meet these and other needs, recombinant nucleic acids (e.g., recombinant herpes virus genomes) encoding one or more CFTR polypeptides for use in viruses (e.g., herpes viruses), pharmaceutical compositions and formulations, medicines and / or methods useful for treating CFTR deficiencies in a subject in need thereof and / or for providing prophylactic, palliative or therapeutic relief of one or more signs or symptoms of a chronic lung disease, such as cystic fibrosis, are provided here.

[008] The present inventors have shown that the recombinant viruses described herein were able to effectively transduce airway epithelial cells derived from a CF patient and successfully express their encoded exogenous human CFTR polypeptides (see, for example, Example 2). Furthermore, the present inventors have shown that the recombinant viruses described herein express full-length functional human CFTR that was adequately transported to the plasma membrane (see, for example, Example 2). In addition, the present inventors have demonstrated that the recombinant viruses described herein rescued the disease phenotype in clinically relevant 3D organotypic cultures prepared from biopsies harvested from multiple CF patients harboring various underlying CFTR mutations (see, for example, Example 3).Furthermore, the present inventors have demonstrated that recombinant HSV vectors can be administered to the lungs of immunocompetent animals via multiple routes and, moreover, that a non-invasive inhaled route of administration expresses similar levels of an encoded transgene while inducing less cellular invasion of the lungs (see, for example, Example 4). Without wishing to be limited by... Petition 870260061885, dated 06 / 24 / 2026, p.17 / 151 / 101 theory, it is believed that increasing, raising and / or supplementing CFTR polypeptide levels in one or more cells (e.g., one or more airway epithelial cells and / or one or more submucosal gland cells) of an individual in need thereof by administering one or more of the nucleic acids, viruses, medications and / or recombinant compositions described herein will: 1) reduce or prevent mucus buildup in one or more organs (e.g., the lungs) of the individual; 2) reduce or prevent airway obstruction in the individual; 3) reduce or prevent chronic bacterial infections and / or associated chronic inflammation in the individual's lungs; 4) reduce or prevent bronchiectasis in the individual; 5) reduce, inhibit or treat progressive lung destruction in the individual; and / or 6) provide prophylactic, palliative, or therapeutic relief of one or more signs or symptoms of a chronic lung disease (e.g., cystic fibrosis, COPD, etc.).

[009] Therefore, certain aspects of the present disclosure relate to a recombinant herpesvirus genome comprising one or more polynucleotides encoding a cystic fibrosis transmembrane conductance regulator (CFTR) polypeptide. In some embodiments, the recombinant herpesvirus genome is replication-competent. In some embodiments, the recombinant herpesvirus genome is replication-defective. In some embodiments that may be combined with any of the foregoing embodiments, the recombinant herpesvirus genome comprises one or more polynucleotides encoding the CFTR polypeptide at one or more viral gene loci.In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes virus genome is selected from a recombinant herpes simplex virus genome, a recombinant varicella-zoster virus genome, a recombinant human cytomegalovirus genome, or a genome of... Petition 870260061885, dated 06 / 24 / 2026, page 18 / 151 / 101 recombinant herpesvirus 6A, a recombinant herpesvirus genome, a recombinant herpesvirus 6B genome, a recombinant herpesvirus 7 genome, a recombinant Kaposi's sarcoma-associated herpesvirus genome, and any combinations or derivatives thereof.

[0010] In some embodiments that can be combined with any of the previous embodiments, the CFTR polypeptide is a human CFTR polypeptide. In some embodiments that can be combined with any of the previous embodiments, the CFTR polypeptide comprises a sequence with at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 6. In some embodiments that can be combined with any of the previous embodiments, the CFTR polypeptide comprises a sequence with at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 5.In some embodiments, the CFTR polypeptide comprises the amino acid sequence SEQ ID NO: 5.

[0011] In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes virus genome is a recombinant herpes simplex virus genome. In some embodiments, the recombinant herpes simplex virus genome is a recombinant herpes simplex virus type 1 (HSV-1) genome, a recombinant herpes simplex virus type 2 (HSV-2) genome, or any derivatives thereof. In some embodiments, the recombinant herpes simplex virus genome is a recombinant HSV-1 genome. Petition 870260061885, dated 06 / 24 / 2026, p. 19 / 151 / 101

[0012] In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation. In some embodiments, the inactivation mutation is in a herpes simplex virus gene. In some embodiments, the inactivation mutation is a deletion of the coding sequence of the herpes simplex virus gene. In some embodiments, the herpes simplex virus gene is selected from Infected Cell Protein (ICP) 0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27, ICP47, thymidine kinase (tk), Single Long Region (UL) 41, and UL55. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in one or both copies of the ICP4 gene.In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP22 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the UL41 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in one or both copies of the ICP0 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP27 gene. In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP47 gene.In some modalities that can be combined with any of the previous modalities, the genome of... Petition 870260061885, dated 06 / 24 / 2026, page 20 / 151 / 101. The recombinant herpes simplex virus comprises an inactivation mutation in the UL55 gene. In some embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises an inactivation mutation in the Hinge region. In some embodiments, the genome of the recombinant herpes simplex virus comprises a deletion of the Hinge region.

[0013] In some embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises one or more polynucleotides encoding the CFTR polypeptide at one or more viral gene loci. In some embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises one or more polynucleotides encoding the CFTR polypeptide at one or both of the viral gene loci ICP4. In some embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises one or more polynucleotides encoding the CFTR polypeptide at the viral gene locus ICP22.In some embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises one or more polynucleotides encoding the CFTR polypeptide at the UL41 viral gene locus. In some embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises one or more polynucleotides encoding the CFTR polypeptide at one or both of the ICP0 viral gene loci. In some embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises one or more polynucleotides encoding the CFTR polypeptide at the ICP27 viral gene locus. In some... Petition 870260061885, dated 06 / 24 / 2026, page 21 / 151 / 101 embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises one or more polynucleotides encoding the CFTR polypeptide at the ICP47 viral gene locus. In some embodiments that can be combined with any of the previous embodiments, the genome of the recombinant herpes simplex virus comprises one or more polynucleotides encoding the CFTR polypeptide at the UL55 viral gene locus.

[0014] In some embodiments that can be combined with any of the previous embodiments, the recombinant herpesvirus genome has reduced cytotoxicity when introduced into a target cell compared to a corresponding wild-type herpesvirus genome. In some embodiments, the cell is a human cell. In some embodiments that can be combined with any of the previous embodiments, the target cell is a respiratory tract cell. In some embodiments that can be combined with any of the previous embodiments, the target cell is an airway epithelial cell or a submucosal gland cell.

[0015] Other aspects of the present disclosure relate to a herpes virus comprising any of the recombinant herpes virus genomes described herein. In some aspects, the herpes virus is competent for replication. In some embodiments, the herpes virus is defective in replication. In some embodiments that can be combined with any of the foregoing embodiments, the herpes virus has reduced cytotoxicity compared to a corresponding wild-type herpes virus. In some embodiments, the herpes virus has reduced cytotoxicity when introduced into a target cell compared to a corresponding wild-type herpes virus. In some embodiments, the cell is a human cell. In some embodiments that can be combined with any of the embodiments Petition 870260061885, dated 06 / 24 / 2026, p. 22 / 151 / 101. In previous embodiments, the target cell is a respiratory tract cell. In some embodiments that can be combined with any of the previous embodiments, the target cell is an airway epithelial cell or a submucosal gland cell. In some embodiments that can be combined with any of the previous embodiments, the herpes virus is selected from a herpes simplex virus, a varicella-zoster virus, a human cytomegalovirus, a herpesvirus 6A, a herpesvirus 6B, a herpesvirus 7, a Kaposi's sarcoma-associated herpesvirus, and any combinations or derivatives thereof. In some embodiments that can be combined with any of the previous embodiments, the herpes virus is a herpes simplex virus. In some forms, the herpes simplex virus is HSV-1, HSV-2, or any derivative thereof. In some forms, the herpes simplex virus is HSV-1.

[0016] Other aspects of this disclosure relate to a pharmaceutical composition comprising any of the recombinant herpesvirus genomes described herein and / or any of the herpesviruses described herein and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is suitable for topical, transdermal, subcutaneous, intradermal, oral, intranasal, intratracheal, sublingual, buccal, rectal, vaginal, inhalation, intravenous, intra-arterial, intramuscular, intracardiac, intraosseous, intraperitoneal, transmucosal, intravitreal, subretinal, intra-articular, periarticular, local and / or epicutaneous use. In some embodiments, the pharmaceutical composition is suitable for oral, intranasal, intratracheal and / or inhalation administration. In some embodiments, the pharmaceutical composition is suitable for inhalation administration. In some embodiments, the pharmaceutical composition is suitable for non-invasive inhalation administration.In some embodiments, the pharmaceutical composition is suitable for use in a dry powder inhaler, a pressurized metered-dose inhaler, a soft mist inhaler, a. Petition 870260061885, dated 06 / 24 / 2026, page 23 / 151 / 101 nebulizer, an electro-hydrodynamic aerosol device or any combination thereof. In some embodiments, the pharmaceutical composition is suitable for nebulization (e.g., using a vibrating mesh nebulizer). In some embodiments that can be combined with any of the previous embodiments, the pharmaceutical composition comprises a phosphate buffer. In some embodiments that can be combined with any of the previous embodiments, the pharmaceutical composition comprises glycerol. In some embodiments that can be combined with any of the previous embodiments, the pharmaceutical composition comprises a lipid carrier. In some embodiments that can be combined with any of the previous embodiments, the pharmaceutical composition comprises a nanoparticle carrier.

[0017] Other aspects of this disclosure relate to the use of any of the recombinant nucleic acids, herpes viruses and / or pharmaceutical compositions described herein as a medicine.

[0018] Other aspects of this disclosure relate to the use of any of the recombinant nucleic acids, herpes viruses and / or pharmaceutical compositions described herein in therapy.

[0019] Other aspects of this disclosure relate to the use of any of the recombinant nucleic acids, herpes viruses and / or pharmaceutical compositions described herein in the production or manufacture of a medicament for the treatment of one or more signs or symptoms of a CFTR deficiency and / or a chronic lung disease (e.g., cystic fibrosis, COPD, etc.).

[0020] Other aspects of the present disclosure relate to a method of intensifying, increasing, raising and / or supplementing the levels of a CFTR polypeptide in one or more cells of a subject, the method comprising administering to the subject an effective amount of either Petition 870260061885, dated 06 / 24 / 2026, page 24 / 151 / 101 of the recombinant herpes virus genomes described herein, any of the herpes viruses described herein and / or any of the pharmaceutical compositions described herein. In some embodiments, one or more cells are one or more cells of the respiratory tract. In some embodiments, one or more cells are one or more epithelial cells of the airways and / or one or more cells of the submucosal glands. In some embodiments that may be combined with any of the preceding embodiments, the subject suffers from a chronic lung disease. In some embodiments, the chronic lung disease is cystic fibrosis or chronic obstructive pulmonary disease (COPD). In some embodiments that may be combined with any of the preceding embodiments, the subject is a human.In some embodiments that can be combined with any of the above embodiments, the subject's genome comprises a loss-of-function mutation in a CFTR gene. In some embodiments that can be combined with any of the above embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, sublingually, buccally, topically, rectally, by inhalation, transdermally, subcutaneously, intradermally, intravenously, intra-arterially, intramuscularly, intracardiacly, intraosseously, intraperitoneally, transmucosally, vaginally, intravitreally, intraorbitally, subretinally, intra-articularly, periarticularly, locally, and / or epicutaneously to the subject. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, or by inhalation to the subject.In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered to the subject by inhalation. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, or by inhalation to the subject. In some embodiments, the recombinant herpes virus genome, the virus... Petition 870260061885, dated 06 / 24 / 2026, page 25 / 151 / 101 of the herpes virus and / or the pharmaceutical composition are administered using a dry powder inhaler, a pressurized metered-dose inhaler, a soft mist inhaler, a nebulizer, or an electro-hydrodynamic aerosol device. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered via a nebulizer (e.g., a vibrating mesh nebulizer).

[0021] Other aspects of the present disclosure relate to a method of reducing or inhibiting the progressive destruction of the lung in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the recombinant herpes virus genomes described herein, any of the herpes viruses described herein, and / or any of the pharmaceutical compositions described herein. In some embodiments, the subject suffers from a chronic lung disease. In some embodiments, the chronic lung disease is cystic fibrosis or chronic obstructive pulmonary disease (COPD). In some embodiments that may be combined with any of the foregoing embodiments, the subject is a human. In some embodiments that may be combined with any of the foregoing embodiments, the subject's genome comprises a loss-of-function mutation in a CFTR gene.In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, sublingually, buccally, topically, rectally, by inhalation, transdermally, subcutaneously, intradermally, intravenously, intra-arterially, intramuscularly, intracardiacally, intraosseously, intraperitoneally, transmucosally, vaginally, intravitreally, intraorbitally, subretinally, intra-articularly, periarticularly, locally, and / or epicutaneously to the subject. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, or by inhalation to the subject. Petition 870260061885, dated 06 / 24 / 2026, p. 26 / 151 / 101 In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered to the subject by inhalation. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, or by inhalation to the subject. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered using a dry powder inhaler, a pressurized metered-dose inhaler, a soft mist inhaler, a nebulizer, or an electro-hydrodynamic aerosol device. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered via a nebulizer (e.g., a vibrating mesh nebulizer).

[0022] Other aspects of the present disclosure relate to a method of providing prophylactic, palliative or therapeutic relief of one or more signs or symptoms of cystic fibrosis in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the recombinant herpes virus genomes described herein, any of the herpes viruses described herein and / or any of the pharmaceutical compositions described herein. In some embodiments, one or more signs or symptoms of cystic fibrosis are selected from a persistent cough producing thick mucus, thick and sticky mucus that accumulates in the airways, wheezing, shortness of breath, sinusitis, repeated lung infections, inflamed nasal passages, bronchiectasis, nasal polyps, hemoptysis, pneumothorax, pancreatitis, recurrent pneumonia, respiratory failure and any combinations thereof.In some modalities that can be combined with any of the previous modalities, the subject is a human. In some modalities that can be combined with any of the previous modalities, the subject's genome comprises a loss-of-function mutation in a CFTR gene. Petition 870260061885, dated 06 / 24 / 2026, p. 27 / 151 / 101, in some embodiments that may be combined with any of the preceding embodiments, the recombinant herpes virus genome, the herpes virus and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, sublingually, buccally, topically, rectally, via inhalation, transdermally, subcutaneously, intradermally, intravenously, intra-arterially, intramuscularly, intracardiacly, intraosseously, intraperitoneally, transmucosally, vaginally, intravitreously, intraorbitally, subretinally, intra-articularly, periarticularly, locally and / or epicutaneously to the subject. In some embodiments, the recombinant herpes virus genome, the herpes virus and / or the pharmaceutical composition are administered orally, intranasally, intratracheally or by inhalation to the subject. In some modalities, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered to the subject via inhalation.In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, or by inhalation to the subject. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered using a dry powder inhaler, a pressurized metered-dose inhaler, a soft mist inhaler, a nebulizer, or an electro-hydrodynamic aerosol device. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered via a nebulizer (e.g., a vibrating mesh nebulizer).

[0023] Other aspects of the present disclosure relate to a method of providing prophylactic, palliative or therapeutic relief of one or more signs or symptoms of COPD in a subject in need thereof, the method comprising administering to the subject an effective amount of any of the recombinant herpes virus genomes described herein, any of the herpes viruses described herein and / or any of the pharmaceutical compositions described herein. In Petition 870260061885, dated 06 / 24 / 2026, p. 28 / 151 / 101 In some modalities, one or more signs or symptoms of COPD are selected from shortness of breath, wheezing, chest tightness, excess mucus in the lungs, chronic cough, cyanosis, frequent respiratory infections, and any combination thereof. In some modalities that may be combined with any of the above modalities, the subject is a human. In some modalities that may be combined with any of the above modalities, the subject's genome comprises a loss-of-function mutation in a CFTR gene.In some embodiments that can be combined with any of the previous embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, sublingually, buccally, topically, rectally, by inhalation, transdermally, subcutaneously, intradermally, intravenously, intra-arterially, intramuscularly, intracardiacly, intraosseously, intraperitoneally, transmucosally, vaginally, intravitreally, intraorbitally, subretinally, intra-articularly, periarticularly, locally, and / or epicutaneously to the subject. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, or by inhalation to the subject. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered by inhalation to the subject.In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered orally, intranasally, intratracheally, or by inhalation to the subject. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered using a dry powder inhaler, a pressurized metered-dose inhaler, a soft mist inhaler, a nebulizer, or an electro-hydrodynamic aerosol device. In some embodiments, the recombinant herpes virus genome, the herpes virus, and / or the pharmaceutical composition are administered via a... Petition 870260061885, dated 06 / 24 / 2026, p. 29 / 151 / 101 nebulizer (for example, a vibrating mesh nebulizer).

[0024] Other aspects of this disclosure relate to an article of manufacture or kit comprising any of the recombinant herpes virus genomes, herpes viruses, medicaments and / or pharmaceutical compositions described herein and instructions for administering the recombinant herpes virus genome, herpes virus, medicament, or pharmaceutical composition. In some embodiments, the article of manufacture or kit further comprises a device for aerosolizing the recombinant herpes virus genome, herpes virus, medicament and / or pharmaceutical composition. In some embodiments, the device is a dry powder inhaler, a pressurized metered-dose inhaler, a soft mist inhaler, a nebulizer, or an electro-hydrodynamic aerosol device. In some embodiments, the device is a nebulizer (e.g., a vibrating mesh nebulizer). BRIEF DESCRIPTION OF THE DRAWINGS

[0025] FIGS. 1A-1I show schematic diagrams of wild-type and modified herpes simplex virus genomes. FIG. 1A shows a wild-type herpes simplex virus genome. FIG. 1B shows a modified herpes simplex virus genome comprising deletions of the ICP4 coding sequence (both copies), with an expression cassette containing a nucleic acid encoding a human CFTR polypeptide integrated into each of the ICP4 loci. FIG. 1C shows a modified herpes simplex virus genome comprising deletions of the ICP4 coding sequences (both copies) and UL41, with an expression cassette containing a nucleic acid encoding a human CFTR polypeptide integrated into each of the ICP4 loci. Figure 1D shows a modified herpes simplex virus genome comprising deletions of the coding sequences of ICP4 (both copies) and UL41, with an expression cassette containing a nucleic acid encoding a CFTR polypeptide integrated into it. Petition 870260061885, dated 06 / 24 / 2026, page 30 / 151 / 101 locus UL41. FIG. 1E shows a modified herpes simplex virus genome comprising deletions of the coding sequences of ICP4 (both copies) and ICP22, with an expression cassette containing a nucleic acid encoding a human CFTR polypeptide integrated into each of the ICP4 loci. FIG. 1F shows a modified herpes simplex virus genome comprising deletions of the coding sequences of ICP4 (both copies) and ICP22, with an expression cassette containing a nucleic acid encoding a CFTR polypeptide integrated into the ICP22 locus. Figure 1G shows a modified herpes simplex virus genome comprising deletions of the coding sequences of ICP4 (both copies), UL41, and ICP22, with an expression cassette containing a nucleic acid encoding a human CFTR polypeptide integrated into each of the ICP4 loci.FIG. 1H shows a modified herpes simplex virus genome comprising deletions of the coding sequences of ICP4 (both copies), UL41, and ICP22, with an expression cassette containing a nucleic acid encoding a CFTR polypeptide integrated into the UL41 locus. FIG. 1I shows a modified herpes simplex virus genome comprising deletions of the coding sequences of ICP4 (both copies), UL41, and ICP22, with an expression cassette containing a nucleic acid encoding a CFTR polypeptide integrated into the ICP22 locus.

[0026] FIG. 2 shows the expression of human CFTR in primary small airway epithelial cells (SAECs) derived from cystic fibrosis (CF) patients infected at indicated multiplicities of infection (MOIs) with an HSV-CFTR vector, as assessed by qRT-PCR analysis. Sham-infected CF SAECs were used as a negative control. Data are presented as the mean of two replicates ± SEM.

[0027] FIG. 3 shows the expression of human CFTR protein in primary SAECs derived from CF patients infected in MOIs indicated with an HSV-CFTR vector, as assessed by analysis of Petition 870260061885, dated 06 / 24 / 2026, page 31 / 151 / 101 western blot. Simulated infected CF SAECs were used as a negative control. GAPDH was used as a loading control.

[0028] FIGS. 4A-4B show immunofluorescence images representative of human CFTR protein expression in primary CF SAECs with sham infection or HSV-CFTR infection. FIG. 4A shows the dose-dependent increase in human CFTR protein expression after infection of primary CF SAECs with increasing HSV-CFTR MOIs. FIG. 4B shows the relative cellular localization of human CFTR protein in primary CF SAECs infected with HSV-CFTR (MOI 3) or sham infection (MOI 0). DAPI staining was used to visualize the nuclei.

[0029] FIG. 5 shows the functionality of the human CFTR protein in primary SAECs derived from CF patients infected in MOIs indicated with an HSV-CFTR vector, as assessed by a fluorescent dye uptake assay. Sham-infected CF SAECs were used as a negative control. Data are presented as mean ± SEM.

[0030] FIGS. 6A-6C show analyses of intestinal organoids derived from G542X / G542X cystic fibrosis patients (PDOs) infected with HSV-CFTR at the indicated MOIs. Vehicle alone or an HSV vector encoding mCherry (mCherry) were used as negative controls; G418 was used as a positive control. FIG. 6A shows representative brightfield images of G542X / G542X PDOs 24 hours after vehicle treatment or after transduction with HSV-CFTR or HSV-mCherry at an MOI of 10. Vehicle-treated PDOs isolated from a healthy individual (wild type) were included and photographed as a comparator. FIG. 6B shows representative images of calcein-stained organoids and quantification of the mean organoid size before the addition of forskolin (Frsk) (t = 0). Figure 6C shows representative images of stained organoids. Petition 870260061885, dated 06 / 24 / 2026, page 32 / 151 / 101 with calcein and quantification of the average size of organoids 60 minutes after the addition of 2 μM of Frsk (t = 60). *** p <0.001; **** p <0.0001.

[0031] FIGS. 7A-7B show analyses of intestinal organoids derived from patients with cystic fibrosis F508del / F508del (PDOs) infected with HSV-CFTR in the indicated MOIs. Vehicle alone or an HSV vector encoding mCherry (mCherry) were used as negative controls; Orkambi® was used as a positive control. FIG. 7A shows representative images of calcein-stained organoids and quantification of the mean organoid size before the addition of forskolin (Frsk) (t = 0). FIG. 7B shows representative images of calcein-stained organoids and quantification of the mean organoid size 60 minutes after the addition of 2 μM of Frsk (t = 60). *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

[0032] FIGS. 8A-8B show analyses of intestinal organoids derived from patients with W1282X / W1282X cystic fibrosis (PDOs) infected with HSV-CFTR in the indicated MOIs. Vehicle alone or an HSV vector encoding mCherry (mCherry) were used as negative controls. FIG. 8A shows representative images of calcein-stained organoids and quantification of the mean organoid size before the addition of forskolin (Frsk) (t = 0). FIG. 8B shows representative images of calcein-stained organoids and quantification of the mean organoid size 60 minutes after the addition of 2 μM of Frsk (t = 60). *p<0.05; ***p<0.001.

[0033] FIGS. 9A-9B show analyses of intestinal organoids derived from cystic fibrosis F508del / F508del (PDOs) patients infected with HSV-CFTR in the indicated MOIs. Vehicle alone or an HSV vector encoding mCherry (mCherry) were used as negative controls; Orkambi® was used as a positive control. FIG. 9A shows representative images of calcein-stained organoids and quantification of the mean organoid size before the addition of forskolin (Frsk) (t = 0). Petition 870260061885, dated 06 / 24 / 2026, page 33 / 151 / 101 FIG. 9B shows representative images of organoids stained with calcein and the quantification of the average size of the organoids 60 minutes after the addition of 2 μM Frsk (t = 60). ****p<0.0001.

[0034] FIGS. 10A-10C show nucleic acid and mCherry protein analyses in lung and trachea biopsies collected 48 hours after intranasal or intratracheal administration of an HSV vector encoding mCherry (HSV-mCherry) or vehicle control (sham). FIG. 10A shows mCherry transcript levels present in lung and trachea biopsies as assessed by qRT-PCR analysis. Data are presented as the mean of six repeats ± SEM for HSV-mCherry; data are presented as the mean of four repeats ± SEM for the vehicle control. FIG. 10B shows representative immunofluorescence images of mCherry protein expression in lung biopsies after intranasal administration of HSV-mCherry or vehicle control. DAPI staining was used to visualize nuclei; cytokeratin staining was used to visualize epithelial cells.Figure 10C shows immunofluorescence images representative of mCherry protein expression in lung biopsies after intratracheal administration of HSVmCherry or vehicle control. DAPI staining was used to visualize nuclei; cytokeratin staining was used to visualize epithelial cells. DETAILED DESCRIPTION

[0035] The following description presents exemplary methods, parameters, and the like. It should be recognized, however, that such a description is not intended to be a limitation on the scope of this disclosure, but rather is provided as a description of exemplary embodiments. I. General Techniques

[0036] The techniques and procedures described or referenced in this document are generally well understood and commonly employed. Petition 870260061885, dated 06 / 24 / 2026, p. 34 / 151 / 101 using conventional methodology by those versed in the technique, such as, for example, the widely used methodologies described in Sambrook et al., Molecular Cloning: A Laboratory Manual 3rd edition (2001) Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; Current Protocols in Molecular Biology (FM Ausubel, et al. eds., (2003)); the Methods in Enzymology series (Academic Press, Inc.): PCR 2: A Practical Approach (MJ MacPherson, BD Hames and GR Taylor eds. (1995)), Harlow and Lane, eds. (1988); Oligonucleotide Synthesis (MJ Gait, ed., 1984); Methods in Molecular Biology, Human Press; Cell Biology: A Laboratory Notebook (JE Cellis, ed., 1998) Academic Press; Animal Cell Culture (RI Freshney, ed., 1987); Introduction to Cell and Tissue Culture (JP Mather and PE Roberts, 1998) Plenum Press; Cell and Tissue Culture: Laboratory Procedures (A. Doyle, JB Griffiths and DG Newell, eds., 1993-8) J.Wiley and Sons; Gene Transfer Vectors for Mammalian Cells (JM Miller and MP Calos, eds., 1987); PCR: The Polymerase Chain Reaction (Mullis et al., eds., 1994); Short Protocols in Molecular Biology (Wiley and Sons, 1999). II. Definitions

[0037] Before describing the invention in detail, it should be understood that this invention is not limited to particular compositions or biological systems, which may, of course, vary. It should also be understood that the terminology used in this document is for the purpose of describing particular embodiments only and is not intended to be limiting.

[0038] As used in this document, the singular forms a / an, an / an and the / the include the referring plural forms, unless the context clearly indicates otherwise. Thus, for example, reference to a molecule optionally includes a combination of two or more such molecules and similar ones.

[0039] As used in this document, the term and / or includes Petition 870260061885, dated 06 / 24 / 2026, p. 35 / 151 / 101 any and all combinations of one or more of the associated listed items. For example, the term ae / or b may refer to aa alone, b alone, a or b or aeb; the term a, be / or c may refer to aa alone, b alone, c alone, a or b, a or c, b or c, a, b, or c”, “aeb”, “aec”, “bec” or “a, bec”; etc.

[0040] As used in this document, the term about refers to the usual error range for the respective value readily known to those skilled in this technical field. The reference to about a value or parameter includes in this document (and describes) embodiments that refer to that value or parameter, per se.

[0041] It is understood that the aspects and embodiments of the invention described in this document include comprising, consisting of and consisting essentially of aspects and embodiments.

[0042] As used, the terms polynucleotide, nucleic acid sequence, nucleic acid and variations thereof should be generic for polydeoxyribonucleotides (containing 2-deoxy-D-ribose), for polyribonucleotides (containing D-ribose), for any other type of polynucleotide that is an N-glycoside of a purine or pyrimidine base, and for other polymers containing non-nucleotide backbones, provided that the polymers contain nucleobases in a configuration that permits base pairing and base stacking, as found in DNA and RNA. Thus, these terms include known types of nucleic acid sequence modifications, for example, substitution of one or more of the naturally occurring nucleotides by an analog and internucleotide modifications.

[0043] A nucleic acid is operationally linked when it is placed in a functional relationship with another nucleic acid sequence. For example, a promoter or enhancer is operationally linked to a coding sequence if it affects the transcription of the sequence, or a site Petition 870260061885, dated 06 / 24 / 2026, p. 36 / 151 / 101 A ribosome-binding molecule is operationally linked to a coding sequence if it is positioned to facilitate translation. Generally, operationally linked or operably bound means that the DNA or RNA sequences being linked are contiguous.

[0044] As used in this document, the term vector refers to distinct elements that are used to introduce heterologous nucleic acids into cells for their expression or replication. An expression vector includes vectors capable of expressing nucleic acids that are operationally linked to regulatory sequences, such as promoter regions, that are capable of effecting the expression of such nucleic acids. Thus, an expression vector may refer to a DNA or RNA construct, such as a plasmid, a phage, a recombinant virus, or another vector that, after introduction into an appropriate host cell, results in the expression of nucleic acids. Appropriate expression vectors are well known to those skilled in the art and include those that are replicable in eukaryotic cells and those that remain episomal or those that integrate into the host cell genome.

[0045] As used in this document, an open reading frame or ORF refers to a continuous stretch of nucleic acids, DNA or RNA, that encodes a protein or polypeptide. Typically, nucleic acids comprise a translation start signal or initiation codon, such as ATG or AUG, and a termination codon.

[0046] As used in this document, an untranslated region or UTR refers to untranslated nucleic acids at the 5' and / or 3' ends of an open reading frame. The inclusion of one or more UTRs in a polynucleotide can affect post-transcriptional regulation, mRNA stability, and / or polynucleotide translation.

[0047] As used in this document, the term transgene refers to a polynucleotide that is capable of being transcribed into RNA and translated. Petition 870260061885, dated 06 / 24 / 2026, page 37 / 151 / 101 and / or expressed under appropriate conditions after being introduced into a cell. In some embodiments, it confers a desired property to a cell into which it has been introduced or otherwise leads to a desired therapeutic or diagnostic outcome.

[0048] The terms polypeptide, peptide, and protein are used interchangeably herein to refer to a polymer of amino acid residues.

[0049] As used in this document, a subject, host, or individual refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sporting, or pet animals such as dogs, horses, cats, cows, as well as animals used in research such as mice, rats, hamsters, rabbits, and non-human primates, etc. In some modalities, the mammal is a human.

[0050] As used in this document, the terms pharmaceutical formulation or pharmaceutical composition refer to a preparation that is in a form that allows the biological activity of the active ingredient(s) to be effective and that does not contain additional components that are unacceptably toxic to a subject to whom the composition or formulation would be administered. Pharmaceutically acceptable excipients (e.g., vehicles, additives) are those that can reasonably be administered to a mammal to provide an effective dose of the active ingredient(s) employed.

[0051] As used in this document, an effective amount is at least the minimum amount necessary to effect a measurable improvement or prevention of one or more symptoms of a particular disorder. An “effective amount” may vary depending on factors such as the disease state, age, sex, and weight of the patient. A therapeutically effective amount is also one in which any toxic or harmful effects of the components are offset by the therapeutically beneficial effects. For Petition 870260061885, dated 06 / 24 / 2026, page 38 / 151 / 101. For prophylactic use, beneficial or desired outcomes include results such as eliminating or reducing the risk, decreasing the severity, or delaying the onset of the disease, its complications, and intermediate pathological phenotypes that present during the development of the disease. For therapeutic use, beneficial or desired outcomes include clinical results such as decreasing one or more symptoms resulting from the disease, increasing the quality of life of those suffering from the disease, decreasing the dose of other medications used to treat the symptoms of the disease, delaying the progression of the disease, and / or prolonging survival. An effective amount may be administered in one or more administrations. For the purposes of this disclosure, an effective amount of a nucleic acid, virus, and / or recombinant pharmaceutical composition is an amount sufficient to perform prophylactic or therapeutic treatment, directly or indirectly.As understood in a clinical context, an effective amount of a nucleic acid, virus, and / or recombinant pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, an effective amount can be considered in the context of administering one or more therapeutic agents, and a single agent can be considered administered in an effective amount if, in conjunction with one or more other agents, a desirable outcome can be or is achieved.

[0052] As used in this document, treatment refers to clinical intervention designed to alter the natural course of the individual or cell being treated during the course of the clinical pathology. Desirable effects of treatment include slowing the rate of progression of the disease / disorder / defect, improvement or palliation of the disease / disorder / defect state, and remission or improvement of the prognosis. For example, an individual is successfully “treated” if one or more symptoms associated with a chronic lung disease (e.g., cystic fibrosis or COPD) are attenuated or eliminated.

[0053] As used in this document, the term delay Petition 870260061885, dated 06 / 24 / 2026, page 39 / 151 / 101. The progression of a disease / disorder / defect refers to delaying, hindering, slowing down, stabilizing, and / or postponing the development of the disease / disorder / defect (e.g., cystic fibrosis or COPD). This delay may vary over different periods of time, depending on the disease history and / or the individual being treated. As is evident to one skilled in the art, a sufficient or significant delay may, in effect, encompass prevention, where the individual does not develop the disease. III. Recombinant Nucleic Acid Sequence

[0054] Certain aspects of the present disclosure relate to recombinant nucleic acids (e.g., isolated recombinant nucleic acids) comprising one or more (e.g., one or more, two or more, three or more, four or more, five or more, ten or more, etc.) polynucleotides encoding a CFTR polypeptide (e.g., a human CFTR polypeptide). In some embodiments, the recombinant nucleic acid comprises one polynucleotide encoding a CFTR polypeptide. In some embodiments, the recombinant nucleic acid comprises two polynucleotides encoding a CFTR polypeptide. In some embodiments, the recombinant nucleic acid comprises three polynucleotides encoding a CFTR polypeptide.

[0055] In some embodiments, the recombinant nucleic acid is a vector. In some embodiments, the recombinant nucleic acid is a viral vector. In some embodiments, the recombinant nucleic acid is a herpes viral vector. In some embodiments, the recombinant nucleic acid is an amplicon of the herpes simplex virus. In some embodiments, the recombinant nucleic acid is a genome of the recombinant herpes virus. In some embodiments, the recombinant nucleic acid is a genome of the recombinant herpes simplex virus. In some embodiments, the recombinant nucleic acid is a genome of the recombinant herpes simplex virus type 1 (HSV-1). Petition 870260061885, dated 06 / 24 / 2026, page 40 / 151 / 101 Polynucleotides encoding cystic fibrosis transmembrane conductance regulator (CFTR) polypeptides

[0056] In some embodiments, the present disclosure refers to a recombinant nucleic acid comprising one or more polynucleotides comprising the coding sequence of a CFTR gene (for example, a human CFTR gene), or any portions thereof.The sequence of any suitable CFTR gene (including any isoform thereof) known in the art may be encoded by a polynucleotide of the present disclosure, including, for example, a human CFTR gene (see, for example, NCBI Gene ID: 1080; SEQ ID NO: 1 or SEQ ID NO: 3), a chimpanzee CFTR gene (see, for example, NCBI Gene ID: 463674), a mouse CFTR gene (see, for example, NCBI Gene ID: 12638), a rat CFTR gene (see, for example, NCBI Gene ID: 24255), a dog CFTR gene (see, for example, NCBI Gene ID: 492302), a rabbit CFTR gene (see, for example, NCBI Gene ID: 100009471), a cow CFTR gene (see, for example, NCBI Gene ID: 281067), a rhesus monkey CFTR gene (see, for example, NCBI Gene ID: 281067). example, NCBI Gene ID: 574346), etc.In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence of any of the CFTR genes described herein or known in the art (and / or the coding sequences thereof). Methods for identifying homologs / orthologs of the CFTR gene from additional species are known to those skilled in the art, including, for example, the use of a nucleic acid sequence alignment program such as the BLAST® blastn suite. Petition 870260061885, dated 06 / 24 / 2026, p. 41 / 151 / 101

[0057] In some embodiments, a polynucleotide of the present disclosure comprises a codon-optimized variant of any of the CFTR genes described herein or known in the art. In some embodiments, a polynucleotide of the present disclosure comprises a codon-optimized variant of the coding sequence of any of the CFTR genes described herein or known in the art. In some embodiments, the use of a codon-optimized variant of a CFTR gene increases the stability and / or yield of heterologous expression (RNA and / or protein) of the encoded CFTR polypeptide in a target cell (e.g., a human target cell, such as a human airway epithelial cell), compared with the stability and / or yield of heterologous expression of a corresponding wild-type sequence not optimized for codons.Any suitable method known in the art for performing codon optimization of a sequence for expression in one or more target cells (e.g., one or more lung cells) may be used, including, for example, the methods described by Fath et al. (PLoS One. March 3, 2011;6(3): e17596).

[0058] In some embodiments, one or more polynucleotides of the present disclosure comprise the coding sequence of a human CFTR gene.

[0059] In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence of SEQ ID NO: 1. In some embodiments, a polynucleotide of the present disclosure comprises the sequence of SEQ ID NO: 1. Petition 870260061885, dated 06 / 24 / 2026, page 42 / 151 / 101

[0060] In some embodiments, a polynucleotide of the present disclosure comprises a 5' truncation, a 3' truncation, or a fragment of the sequence with SEQ ID NO: 1. In some embodiments, the 5' truncation, 3' truncation, or fragment of the sequence with SEQ ID NO: 1 is a polynucleotide that has at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2250, at least 2500, at least 2750, at least 3000, at least 3250, at least 3500, at least 3750, at least 4000, at least 4250, but less than 4443 consecutive nucleotides of SEQ ID NO: 1.In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the nucleic acid sequence 1-4440 of SEQ ID NO: 1. In some embodiments, a polynucleotide of the present disclosure comprises the nucleic acid sequence 1-4440 of SEQ ID NO: 1.

[0061] In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence of SEQ ID NO: 3. In some embodiments, a polynucleotide of the present disclosure comprises the sequence of SEQ ID Petition 870260061885, dated 06 / 24 / 2026, page 43 / 151 / 101 NO: 3.

[0062] In some embodiments, a polynucleotide of the present disclosure comprises a 5' truncation, a 3' truncation, or a fragment of the sequence with SEQ ID NO: 3. In some embodiments, the 5' truncation, 3' truncation, or fragment of the sequence with SEQ ID NO: 3 is a polynucleotide that has at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2250, at least 2500, at least 2750, at least 3000, at least 3250, at least 3500, at least 3750, at least 4000, at least 4250, but less than 4260 consecutive nucleotides of SEQ ID NO: 3.In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with nucleic acid sequence 1-4257 of SEQ ID NO: 3. In some embodiments, a polynucleotide of the present disclosure comprises nucleic acid sequence 1-4257 of SEQ ID NO: 3.

[0063] In some embodiments, a polynucleotide of the present disclosure comprises the coding sequence of a codon-optimized variant of a human CFTR gene.

[0064] In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, Petition 870260061885, dated 06 / 24 / 2026, page 44 / 151 / 101 at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence with SEQ ID NO: 2. In some embodiments, a polynucleotide of this disclosure comprises the sequence with SEQ ID NO: 2.

[0065] In some embodiments, a polynucleotide of the present disclosure comprises a 5' truncation, a 3' truncation, or a fragment of the sequence with SEQ ID NO: 2. In some embodiments, the 5' truncation, 3' truncation, or fragment of the sequence with SEQ ID NO: 2 is a polynucleotide that has at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1000, at least 1250, at least 1500, at least 1750, at least 2000, at least 2250, at least 2500, at least 2750, at least 3000, at least 3250, at least 3500, at least 3750, at least 4000, at least 4250, but less than 4443 consecutive nucleotides of SEQ ID NO: 2.In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the nucleic acid sequence 1-4440 of SEQ ID NO: 2. In some embodiments, a polynucleotide of the present disclosure comprises the nucleic acid sequence 1-4440 of SEQ ID NO: 2.

[0066] In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, Petition 870260061885, dated 06 / 24 / 2026, p. 45 / 151 / 101 at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the sequence with SEQ ID NO: 4. In some embodiments, a polynucleotide of the present disclosure comprises the sequence with SEQ ID NO: 4.

[0067] In some embodiments, a polynucleotide of the present disclosure comprises a 5' truncation, a 3' truncation, or a fragment of the sequence with SEQ ID NO: 4. In some embodiments, the 5' truncation, 3' truncation, or fragment of the sequence with SEQ ID NO: 4 is a polynucleotide that has at least 25, at least 50, at least 75, at least 100, at least 125, at least 150, at least 175, at least 200, at least 250, at least 300, or at least 350, at least 400, at least 450, at least 500, at least 750, at least 1,000, at least 1,250, at least 1,500, at least 1,750, at least 2,000, at least 2,250, at least less than 2,500, at least 2,750, at least 3,000, at least 3,250, at least 3,500, at least 3,750, at least 4,000, at least 4,250, but less than 4,260 consecutive nucleotides of SEQ ID NO: 4.In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the nucleic acid sequence 1-4257 of SEQ ID NO: 4. In some embodiments, a polynucleotide of the present disclosure comprises the nucleic acid sequence 1-4257 of SEQ ID NO: 4.

[0068] In some embodiments, a polynucleotide of the present Petition 870260061885, dated 06 / 24 / 2026, page 46 / 151 / 101 disclosure comprises a sequence with at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NOS: 1-4. In some embodiments, a polynucleotide of the present disclosure comprises a sequence selected from SEQ ID NOS: 1-4. In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NO: 1 or SEQ ID NO: 2.In some embodiments, a polynucleotide of the present disclosure comprises a sequence selected from SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, a polynucleotide of the present disclosure comprises a sequence with at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least less than 98%, at least 99%, or 100% sequence identity with a nucleic acid sequence selected from SEQ ID NO: 3 or SEQ ID NO: 4. In some embodiments, a polynucleotide of the present disclosure comprises a sequence selected from SEQ ID NO: 3 or SEQ ID NO: 4.

[0069] A polynucleotide of the present disclosure (e.g., encoding a human CFTR polypeptide) may further encode additional coding and non-coding sequences. Examples of additional coding and non-coding sequences may include, but are not limited to, sequences encoding additional polypeptide markers (e.g., encoded in-frame with the CFTR protein in order to produce Petition 870260061885, dated 06 / 24 / 2026, p. 47 / 151 / 101 a fusion protein), introns (e.g., native, modified, or heterologous introns), 5' and / or 3' UTRs (e.g., native, modified, or heterologous 5' and / or 3' UTRs) and the like. Examples of suitable polypeptide markers may include, but are not limited to, any combination of purification tags, such as hist tags, marker tags, maltose-binding protein and glutathione-Stransferase tags, detection tags, such as tags that can be detected photometrically (e.g., green fluorescent protein, red fluorescent protein, etc.) and tags that have detectable enzymatic activity (e.g., alkaline phosphatase, etc.).), labels containing secretory sequences, signal sequences, leader sequences and / or stabilizing sequences, protease cleavage sites (e.g., furin cleavage sites, TEV cleavage sites, thrombin cleavage sites, etc.), and the like. In some embodiments, 5' and / or 3' UTRs enhance the stability, localization, and / or translation efficiency of polynucleotides. In some embodiments, 5' and / or 3' UTRs improve the level and / or duration of protein expression. In some embodiments, 5' and / or 3' UTRs include elements (e.g., one or more miRNA binding sites, etc.) that can block or reduce off-target expression (e.g., inhibit expression in specific cell types (e.g., neuronal cells), at specific times in the cell cycle, at specific developmental stages, etc.). In some embodiments, the 5' and / or 3' UTRs include elements (e.g., one or more miRNA binding sites, etc.).which can enhance CFTR expression in specific cell types.

[0070] In some embodiments, a polynucleotide of the present disclosure (for example, encoding a human CFTR polypeptide) is operationally linked to one or more (for example, one or more, two or more, three or more, four or more, five or more, ten or more, etc.) Petition 870260061885, dated 06 / 24 / 2026, page 48 / 151 / 101 regulatory sequences. The term regulatory sequence includes promoters, enhancers, and other expression control elements (e.g., polyadenylation signals). Any suitable enhancer(s) known in the art may be used, including, for example, enhancer sequences of mammalian genes (such as globin, elastase, albumin, α-fetoprotein, insulin, and the like), enhancer sequences of eukaryotic cell viruses (such as SV40 enhancer on the late side of the origin of replication (bp 100-270), cytomegalovirus early promoter enhancer, polyome enhancer on the late side of the origin of replication, adenovirus enhancers, and the like) and any combinations thereof.Any suitable isolator(s) known in the art may be used, including, for example, herpes simplex virus (HSV) chromatin boundary elements (CTRL / binding / CTCF isolator) CTRL1 and / or CTRL2, chicken hypersensitive site isolator 4 (cHS4), ubiquitous human HNRPA2B1 — CBX3 chromatin opening element (UCOE), the scaffold / matrix attachment region (S / MAR) of the human interferon beta gene (IFNB1), and any combinations thereof.Any suitable promoter (e.g., suitable for transcription in mammalian host cells) known in the art may be used, including, for example, promoters obtained from viral genomes (such as polyomavirus, avian poxvirus, adenovirus (such as Adenovirus 2), bovine papillomavirus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis B virus, Simian Virus 40 (SV40) and the like), promoters of heterologous mammalian genes (such as the actin promoter (e.g., the β-actin promoter), a ubiquitin promoter (e.g., a ubiquitin C (UbC) promoter), a phosphoglycerate kinase (PGK) promoter, an immunoglobulin promoter, heat shock protein promoters and the like), promoters of native and / or homologous mammalian genes (e.g., a promoter of... Petition 870260061885, dated 06 / 24 / 2026, p. 49 / 151 / 101 human CFTR gene), synthetic promoters (such as the CAGG promoter) and any combinations thereof, provided that such promoters are compatible with host cells. Regulatory sequences may include those that direct the constitutive expression of a nucleic acid, as well as tissue-specific and / or inducible or repressible regulatory sequences.

[0071] In some embodiments, a polynucleotide of the present disclosure is operationally linked to one or more heterologous promoters. In some embodiments, one or more heterologous promoters are one or more of the constitutive promoters, tissue-specific promoters, temporal promoters, spatial promoters, inducible promoters, and repressible promoters. In some embodiments, one or more heterologous promoters are one or more of the immediate early promoter of human cytomegalovirus (HCMV), the human elongation factor-1 (EF1) promoter, the human β-actin promoter, the human UbC promoter, the human PGK promoter, the synthetic CAGG promoter, and any combinations thereof. In some embodiments, a polynucleotide of the present disclosure (e.g., encoding a human CFTR polypeptide) is operationally linked to an HCMV promoter.

[0072] In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence (e.g., a transgene encoding) of a collagen alpha-1 (VII) chain polypeptide (COL7). In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence of (e.g., a transgene encoding) of a Lysyl hydroxylase 3 (LH3) polypeptide. In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence of (e.g., a transgene encoding) of a type I keratin cytoskeletal polypeptide 17 (KRT17). In some embodiments, a polynucleotide of the present disclosure does not Petition 870260061885, dated 06 / 24 / 2026, page 50 / 151 / 101 comprises the coding sequence (e.g., a transgene encoding) a transglutaminase (TGM) polypeptide (e.g., a human transglutaminase polypeptide, such as a human TGM1 polypeptide and / or a human TGM5 polypeptide). In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence of (e.g., a transgene encoding) a cosmetic protein (e.g., collagen proteins, fibronectins, elastins, lumicans, vitronectins / vitronectin receptors, laminins, neuromodulators, fibrillins, additional dermal extracellular matrix proteins, etc.). In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence of (e.g., a transgene encoding) an antibody (e.g., a full-length antibody, an antibody fragment, etc.).In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence of (e.g., a transgene encoding) a serine protease inhibitor (SPINK) Kazal-like polypeptide (e.g., a human SPINK polypeptide, such as a SPINK5 polypeptide). In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence of (e.g., a transgene encoding) a filaggrin or filaggrin 2 polypeptide (e.g., a human filaggrin or filaggrin 2 polypeptide). In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence of (e.g., a transgene encoding) a collagen alpha-1 (VII) chain polypeptide, a lysyl hydroxylase 3 polypeptide, a type I keratin cytoskeletal polypeptide 17 and / or any chimeric polypeptides thereof.In some embodiments, a polynucleotide of the present disclosure does not comprise the coding sequence of (for example, a transgene encoding) a collagen alpha-1 (VII) chain polypeptide, a lysyl hydroxylase 3 polypeptide, a. Petition 870260061885, dated 06 / 24 / 2026, page 51 / 151 / 101 cytoskeletal polypeptide 17 of keratin type I, a transglutaminase (TGM) polypeptide, a filaggrin polypeptide, a cosmetic protein, an antibody, a SPINK polypeptide and / or any chimeric polypeptides thereof. Cystic fibrosis transmembrane conductance regulating polypeptides (CFTR)

[0073] In some embodiments, the present disclosure refers to one or more polynucleotides encoding a CFTR polypeptide (for example, a human CFTR polypeptide) or any portions thereof. Any suitable CFTR polypeptide known in the art can be encoded by a polynucleotide of the present disclosure, including, for example, a human CFTR polypeptide (see, for example, UniProt accession number P13569; SEQ ID NO: 5 or SEQ ID NO: 6), a chimpanzee CFTR polypeptide (see, for example, UniProt accession number Q2QLE5), a mouse CFTR polypeptide (see, for example, UniProt accession number P26361), a rat CFTR polypeptide (see, for example, UniProt accession number P34158), a rabbit CFTR polypeptide (see, for example, UniProt accession number Q00554), a rhesus monkey CFTR polypeptide (see, for example, UniProt accession number Q00553), etc.In some embodiments, a CFTR polypeptide of the present disclosure comprises a sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity with the amino acid sequence of any of the CFTR polypeptides described herein or known in the art. Methods for identifying homologs / orthologs of CFTR polypeptides from additional species are known to those skilled in the art, including, for example, the use of... Petition 870260061885, dated 06 / 24 / 2026, page 52 / 151 / 101 an amino acid sequence alignment program, such as BLAST® blastp suite or OrthoDB.

[0074] In some embodiments, a CFTR polypeptide of the present disclosure is a human CFTR polypeptide.

[0075] In some embodiments, a polynucleotide encoding a human CFTR polypeptide is a polynucleotide encoding a polypeptide comprising an amino acid sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity with the sequence of SEQ ID NO: 5. In some embodiments, a polynucleotide encoding a human CFTR polypeptide is a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 5.

[0076] In some embodiments, a polynucleotide encoding a human CFTR polypeptide is a polynucleotide encoding an N-terminal truncation, a C-terminal truncation, or a fragment of the amino acid sequence of SEQ ID NO: 5. N-terminal truncations, C-terminal truncations, or fragments may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900, at least 950, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, but less than 1480, consecutive amino acids of SEQ ID NO: 5. Petition 870260061885, dated 06 / 24 / 2026, page 53 / 151 / 101

[0077] In some embodiments, a polynucleotide encoding a human CFTR polypeptide is a polynucleotide encoding a polypeptide comprising an amino acid sequence with at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identity with the sequence of SEQ ID NO: 6. In some embodiments, a polynucleotide encoding a human CFTR polypeptide is a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 6.

[0078] In some embodiments, a polynucleotide encoding a human CFTR polypeptide is a polynucleotide encoding an N-terminal truncation, a C-terminal truncation, or a fragment of the amino acid sequence of SEQ ID NO: 6. N-terminal truncations, C-terminal truncations, or fragments may comprise at least 10, at least 12, at least 14, at least 16, at least 18, at least 20, at least 30, at least 40, at least 50, at least 75, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900, at least 950, at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, but less than 1419, consecutive amino acids of SEQ ID NO: 6.

[0079] In some embodiments, a polynucleotide of the present disclosure encoding a CFTR polypeptide is a polynucleotide encoding a polypeptide comprising a sequence with at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, Petition 870260061885, dated 06 / 24 / 2026, page 54 / 151 / 101 at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 6. In some embodiments, a polynucleotide of the present disclosure encoding a CFTR polypeptide is a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO: 6.

[0080] In some embodiments, a polynucleotide of the present disclosure encoding a CFTR polypeptide (e.g., a human CFTR polypeptide) expresses the CFTR polypeptide when the polynucleotide is distributed to one or more target cells of a subject (e.g., one or more cells of the subject's airways and / or lungs). In some embodiments, the expression of the CFTR polypeptide (e.g., a human CFTR polypeptide) intensifies, elevates, increases, and / or supplements the levels, function, and / or activity of a CFTR polypeptide in one or more target cells of a subject (e.g., compared to before the expression of the CFTR polypeptide). In some embodiments, the expression of the CFTR polypeptide (e.g., a human CFTR polypeptide) reduces mucus secretion by one or more cells and / or in one or more organs (e.g., the lungs) of the subject (e.g., compared to before the expression of the CFTR polypeptide).In some embodiments, expression of the CFTR polypeptide (e.g., a human CFTR polypeptide) reduces and / or inhibits mucus accumulation in one or more organs (e.g., the lungs) of the subject (e.g., compared to before CFTR polypeptide expression). In some embodiments, expression of the CFTR polypeptide (e.g., a human CFTR polypeptide) reduces, prevents, or treats airway obstruction in a subject (e.g., compared to before CFTR polypeptide expression). In some embodiments, expression of the CFTR polypeptide (e.g., a CFTR polypeptide) Petition 870260061885, dated 06 / 24 / 2026, page 55 / 151 / 101 human) reduces, prevents, or treats chronic bacterial infections and / or associated chronic inflammation in a subject's lungs (e.g., compared to before CFTR polypeptide expression). In some embodiments, CFTR polypeptide expression (e.g., a human CFTR polypeptide) reduces, inhibits, prevents, or treats bronchiectasis in a subject (e.g., compared to before CFTR polypeptide expression). In some embodiments, CFTR polypeptide expression (e.g., a human CFTR polypeptide) reduces, inhibits, prevents, or treats progressive lung destruction in a subject (e.g., compared to before CFTR polypeptide expression).In some embodiments, expression of the CFTR polypeptide (e.g., a human CFTR polypeptide) provides prophylactic, palliative, or therapeutic relief of a chronic lung disease (e.g., cystic fibrosis, chronic obstructive pulmonary disease) in a subject (e.g., compared to before CFTR polypeptide expression). In some embodiments, expression of the CFTR polypeptide (e.g., a human CFTR polypeptide) provides prophylactic, palliative, or therapeutic relief of one or more signs or symptoms of cystic fibrosis in a subject (e.g., compared to before CFTR polypeptide expression). Recombinant Nucleic Acid Sequence

[0081] In some embodiments, the present disclosure refers to recombinant nucleic acids comprising any one or more of the polynucleotides described herein. In some embodiments, the recombinant nucleic acid is a vector (e.g., an expression vector, a display vector, etc.). In some embodiments, the vector is a DNA vector or an RNA vector. Generally, vectors suitable for maintaining, propagating, and / or expressing polynucleotides to produce one or more polypeptides in a subject may be used. Examples of suitable vectors may include, for example, Petition 870260061885, dated 06 / 24 / 2026, page 56 / 151 / 101 example, plasmids, cosmids, episomes, transposons, and viral vectors (e.g., adenoviral vectors, adeno-associated viral vectors, vaccinia viral vectors, Sindbis viral vectors, measles vectors, herpes viral vectors, lentiviral vectors, retroviral vectors, etc.). In some embodiments, the vector is a herpes viral vector. In some embodiments, the vector is capable of autonomous replication in a host cell. In some embodiments, the vector is incapable of autonomous replication in a host cell. In some embodiments, the vector can integrate into host DNA. In some embodiments, the vector cannot integrate into host DNA (e.g., it is episomal).Methods for producing vectors containing one or more polynucleotides of interest are well known to those skilled in the art, including, for example, chemical synthesis or artificial manipulation of isolated nucleic acid segments (e.g., by genetic engineering techniques).

[0082] In some embodiments, a recombinant nucleic acid of the present disclosure is an amplicon of herpes simplex virus (HSV). Herpes virus amplicons, including the structural features and methods for making them, are generally known to those skilled in the art (see, for example, de Silva S. and Bowers W. “Herpes Virus Amplicon Vectors”. Viruses 2009, 1, 594-629). In some embodiments, the herpes simplex virus amplicon is an HSV-1 amplicon. In some embodiments, the herpes simplex virus amplicon is a hybrid HSV-1 amplicon. Examples of HSV-1 hybrid amplicons may include, but are not limited to, HSV / AAV hybrid amplicons, HSV / EBV hybrid amplicons, HSV / EBV / RV hybrid amplicons, and / or HSV / Sleeping Beauty hybrid amplicons. In some embodiments, the amplicon is an HSV / AAV hybrid amplicon. In some embodiments, the amplicon is an HSV / Sleeping Beauty hybrid amplicon.

[0083] In some embodiments, a recombinant nucleic acid of Petition 870260061885, dated 06 / 24 / 2026, page 57 / 151 / 101, the present disclosure is a recombinant herpesvirus genome. The recombinant herpesvirus genome may be a recombinant genome of any member of the Herpesviridae family of DNA viruses known in the art, including, for example, a recombinant herpes simplex virus genome, a recombinant varicella-zoster virus genome, a recombinant human cytomegalovirus genome, a recombinant herpesvirus 6A genome, a recombinant herpesvirus 6B genome, a recombinant herpesvirus 7 genome, a recombinant Kaposi's sarcoma-associated herpesvirus genome, and any combinations or derivatives thereof.As used in this document, an inactivating mutation may refer to any mutation that results in a gene or regulon product (RNA or protein) having reduced, undetectable, or eliminated quantity and / or function (e.g., compared to a corresponding sequence without the inactivating mutation). Examples of inactivating mutations may include, but are not limited to, deletions, insertions, point mutations, and rearrangements in transcription control sequences (promoters, enhancers, insulators, etc.) and / or coding sequences of a given gene or regulon. Any method suitable for measuring the quantity of a gene or regulon product known in the art may be used, including, for example, qPCR, Northern blots, RNAseq, Western blots, ELISAs, etc.In some embodiments, the recombinant herpesvirus genome comprises one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) inactivating mutations. In some embodiments, one or more inactivating mutations are in one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) herpesvirus genes. In some embodiments, the recombinant herpesvirus genome is attenuated (e.g., in... Petition 870260061885, dated 06 / 24 / 2026, p. 58 / 151 / 101 comparison with a corresponding wild herpes virus genome). In some embodiments, the recombinant herpes virus genome is competent for replication. In some embodiments, the recombinant herpes virus genome is defective in replication.

[0084] In some embodiments, the recombinant nucleic acid is a recombinant herpes simplex virus (HSV) genome. In some embodiments, the recombinant herpes simplex virus genome is a herpes simplex virus type 1 (HSV-1) genome, a recombinant herpes simplex virus type 2 (HSV-2) genome, or any derivatives thereof. In some embodiments, the recombinant herpes simplex virus genome comprises one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) inactivating mutations. In some embodiments, the one or more inactivating mutations are in one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) herpes simplex virus genes.In some embodiments, the genome of the recombinant herpes simplex virus is attenuated (e.g., compared to a corresponding wild-type herpes simplex virus genome). In some embodiments, the genome of the recombinant herpes simplex virus is competent for replication. In some embodiments, the genome of the recombinant herpes simplex virus is defective in replication.

[0085] In some embodiments, the recombinant herpes simplex virus genome is a recombinant HSV-1 genome. In some embodiments, the recombinant HSV-1 genome may be from any HSV-1 strain known in the art, including, for example, strains 17, Ty25, R62, S25, Ku86, S23, R11, Ty148, Ku47, H166syn, 1319-2005, F-13, M-12, 90237, F-17, KOS, 3083-2008, F12g, L2, CD38, H193, M-15, India 2011, Petition 870260061885, dated 06 / 24 / 2026, page 59 / 151 / 101 0116209, F-11I, 66-207, 2762, 369-2007, 3355, MacIntyre, McKrae, 7862, 7hse, HF10, 1394,2005, 270-2007, OD4, SC16, M-19, 4J1037, 5J1060, J1060, KOS79, 132-1988, 160-1982, H166, 2158-2007, RE, 78326, F18g, F11, 1722010, H129, F, E4, CJ994, F14g, E03, E22, E10, E06, E11, E25, E23, E35, E15, E07, E12, E14, E08, E19, E13, ATCC 2011, etc. (see, for example, Bowen et al. J Virol. April 3, 2019;93(8)). In some embodiments, the recombinant HSV-1 genome is from the KOS strain. In some embodiments, the recombinant HSV-1 genome is not from the McKrae strain. In some embodiments, the recombinant HSV-1 genome is attenuated. In some embodiments, the recombinant HSV-1 genome is replication-competent. In some embodiments, the recombinant HSV-1 genome is replication-defective.In some embodiments, the recombinant HSV-1 genome comprises one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) inactivating mutations. In some embodiments, one or more inactivating mutations are in one or more (e.g., one or more, two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, ten or more, etc.) HSV-1 genes.

[0086] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in at least one, at least two, at least three, at least four, at least five, at least six, at least seven, or all eight of the infected cellular protein (ICP) 0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27, ICP47, thymidine kinase (tk), herpes simplex virus genes of the long single region (UL) 41 and / or UL55. In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivating mutation (e.g., it is capable of expressing) the herpes simplex virus gene ICP0 (one or both copies). In some embodiments, the recombinant herpes simplex virus genome does not comprise a Petition 870260061885, dated 06 / 24 / 2026, page 60 / 151 / 101 inactivation mutation (e.g., is capable of expression) in the herpes simplex virus gene ICP27. In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivation mutation (e.g., is capable of expression) in the herpes simplex virus gene ICP47. In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivation mutation (e.g., is capable of expression) in the herpes simplex virus genes ICP0 (one or both copies), ICP27, and / or ICP47. In some embodiments, the recombinant herpes simplex virus genome does not comprise an inactivation mutation in the Hinge region. In some embodiments, the genome of the recombinant herpes simplex virus does not comprise an inactivating mutation in ICP34.5 (one or both copies) and / or herpes simplex virus ICP47 genes (e.g., to prevent the production of an immunostimulatory virus).In some embodiments, the genome of the recombinant herpes simplex virus does not comprise an inactivating mutation in the herpes simplex virus gene ICP34.5 (one or both copies). In some embodiments, the genome of the recombinant herpes simplex virus does not comprise an inactivating mutation in the herpes simplex virus gene ICP47. In some embodiments, the genome of the recombinant herpes simplex virus does not comprise an inactivating mutation in the herpes simplex virus genes ICP34.5 (one or both copies) and ICP47. In some embodiments, the genome of the recombinant herpes simplex virus is not oncolytic.

[0087] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP0 gene (one or both copies) and also comprises an inactivating mutation in ICP4 (one or both copies), ICP22, ICP27, ICP47, UL41, Petition 870260061885, dated 06 / 24 / 2026, p. 61 / 151 / 101 and / or UL55 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 gene (one or both copies) and an inactivation mutation in the ICP4 gene (one or both copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 gene (one or both copies) and an inactivation mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 gene (one or both copies) and an inactivation mutation in the UL41 gene. In some forms, the genome of the recombinant herpes simplex virus comprises an inactivating mutation in the ICP0 gene (one or both copies), an inactivating mutation in the ICP4 gene (one or both copies), and an inactivating mutation in the ICP22 gene.In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 gene (one or both copies), an inactivation mutation in the ICP4 gene (one or both copies), and an inactivation mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 gene (one or both copies), an inactivation mutation in the ICP22 gene, and an inactivation mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 gene (one or both copies), an inactivation mutation in the ICP4 gene (one or both copies), an inactivation mutation in the ICP22 gene, and an inactivation mutation in the UL41 gene. In some forms, the inactivating mutation is a deletion of the coding sequence of the ICP0 (one or both copies), ICP4 (one or both copies), ICP22, and / or UL41 genes.In some forms, the genome of the recombinant herpes simplex virus also includes an inactivating mutation in the ICP27, ICP47, and / or UL55 genes. Petition 870260061885, dated 06 / 24 / 2026, p. 62 / 151 / 101

[0088] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP4 gene (one or both copies). In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP4 gene (one or both copies) and also comprises an inactivation mutation in the ICP0 (one or both copies), ICP22, ICP27, ICP47, UL41, and / or UL55 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP4 gene (one or both copies) and an inactivation mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP4 gene (one or both copies) and an inactivation mutation in the UL41 gene.In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP4 gene (one or both copies), an inactivating mutation in the ICP22 gene, and an inactivating mutation in the UL41 gene. In some embodiments, the inactivating mutation is a deletion of the gene coding sequence, ICP4 (one or both copies), ICP22, and / or UL41. In some embodiments, the recombinant herpes simplex virus genome further comprises an inactivating mutation in the ICP0 genes (one or both copies), ICP27, ICP47, and / or UL55.

[0089] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene and also comprises an inactivating mutation in ICP0 (one or both copies), ICP4 (one or both copies), ICP27, ICP47, UL41, and / or UL55 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the ICP22 gene and an inactivating mutation in the UL41 gene. In some embodiments, the mutation Petition 870260061885, dated 06 / 24 / 2026, p. 63 / 151 / 101 of inactivation is a deletion of the coding sequence of the ICP22 and / or UL41 genes. In some embodiments, the genome of the recombinant herpes simplex virus also comprises an inactivation mutation in the ICP0 (one or both copies), ICP4 (one or both copies), ICP27, ICP47 and / or UL55 genes.

[0090] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP27 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP27 gene and also comprises an inactivation mutation in ICP0 (one or both copies), ICP4 (one or both copies), ICP22, ICP47, UL41, and / or UL55 genes. In some embodiments, the inactivation mutation is a deletion of the coding sequence of the ICP27 gene.

[0091] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP47 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP47 gene and also comprises an inactivation mutation in ICP0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27, UL41, and / or UL55 genes. In some embodiments, the inactivation mutation is a deletion of the coding sequence of the ICP47 gene.

[0092] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the UL41 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the UL41 gene and also comprises an inactivation mutation in the ICP0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27, ICP47, and / or UL55 genes. In some embodiments, the inactivation mutation is a deletion of the coding sequence of the UL41 gene. Petition 870260061885, dated 06 / 24 / 2026, p. 64 / 151 / 101

[0093] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL55 gene. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation in the UL55 gene and also comprises an inactivating mutation in the ICP0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27, ICP47, and / or UL41 genes. In some embodiments, the inactivating mutation is a deletion of the coding sequence of the UL55 gene.

[0094] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivating mutation (e.g., a deletion) of the internal repeat (Hinge) region comprising the long internal repeat (IRL) and short internal repeat (IRS) regions. In some embodiments, inactivation (e.g., deletion) of the Hinge region eliminates one copy of each of the ICP4 and ICP0 genes. In some embodiments, inactivation (e.g., deletion) of the Hinge region further inactivates (e.g., deletes) the promoter for the ICP22 and ICP47 genes. If desired, the expression of one or both genes can be restored by inserting an immediate early promoter into the genome of the recombinant herpes simplex virus (see, for example, Hill et al. (1995). Nature 375(6530): 411-415; Goldsmith et al. (1998). J Exp Med 187(3): 341-348).Without wishing to be limited by theory, it is believed that inactivation (e.g., deletion) of the common region may contribute to the stability of the recombinant herpes simplex virus genome and / or allow the recombinant herpes simplex virus genome to accommodate more transgenes and / or larger transgenes.

[0095] In some embodiments, the genome of the recombinant herpes simplex virus comprises an inactivating mutation in the ICP4 (one or both copies), ICP22, and ICP27 genes. In some embodiments, the genome of the recombinant herpes simplex virus comprises a mutation Petition 870260061885, dated 06 / 24 / 2026, p. 65 / 151 / 101 concerning inactivation mutations in the ICP4 (one or both copies), ICP27, and UL55 genes. In some embodiments, the recombinant herpes simplex virus genome also comprises an inactivation mutation in the ICP4 (one or both copies), ICP22, ICP27, ICP47, and / or UL55 genes. In some embodiments, the inactivation mutation in the ICP4 (one or both copies), ICP27, and / or UL55 genes is a deletion of the coding sequence of the ICP4 (one or both copies), ICP27, and / or UL55 genes. In some embodiments, the inactivating mutation in the ICP22 and ICP47 genes is a deletion in the promoter region of the ICP22 and ICP47 genes (e.g., the ICP22 and ICP47 coding sequences are intact but not transcriptionally active). In some embodiments, the recombinant herpes simplex virus genome comprises a deletion in the coding sequence of the ICP4 (one or both copies), ICP27, and UL55 genes, and a deletion in the promoter region of the ICP22 and ICP47 genes.In some forms, the genome of the recombinant herpes simplex virus also includes an inactivating mutation in the ICP0 genes (one or both copies) and / or UL41 genes.

[0096] In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 (one or both copies) and ICP4 (one or both copies) genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 (one or both copies), ICP4 (one or both copies), and ICP22 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 (one or both copies), ICP4 (one or both copies), ICP22, and ICP27 genes. In some embodiments, the recombinant herpes simplex virus genome comprises an inactivation mutation in the ICP0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27, and UL55 genes. In some embodiments, the inactivation mutation in the ICP0 (one or both copies), ICP4 (one or both copies) genes Petition 870260061885, dated 06 / 24 / 2026, p. 66 / 151 / 101 copies), ICP22, ICP27 and / or UL55 comprises a deletion of the coding sequence of ICP0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27 and / or UL55. In some embodiments, the recombinant herpes simplex virus genome further comprises an inactivation mutation in the ICP47 and / or UL41 genes.

[0097] In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure at one, two, three, four, five, six, seven or more viral gene loci. Examples of suitable viral loci may include, without limitation, the herpes simplex viral gene loci ICP0 (one or both copies), ICP4 (one or both copies), ICP22, ICP27, ICP47, tk, UL41 and / or UL55. In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within one or both viral ICP4 gene loci (e.g., a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at one or both ICP4 loci).In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within the viral ICP22 gene locus (e.g., a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at the ICP22 locus). In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within the viral UL41 gene locus (e.g., a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at the UL41 locus). In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure in one or both of the viral ICP0 gene loci (e.g., a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at one or both ICP0 loci). In some embodiments, a herpes simplex virus genome... Petition 870260061885, dated 06 / 24 / 2026, page 67 / 151 / 101. A recombinant herpes simplex virus comprises one or more polynucleotides of the present disclosure within the viral ICP27 gene locus (for example, a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at the ICP27 locus). In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within the viral ICP47 gene locus (for example, a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at the ICP47 locus).In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure at one or both loci of the viral ICP4 gene and one or more polynucleotides of the present disclosure within the locus of the viral ICP22 gene (for example, a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at one or both ICP4 loci, and a polynucleotide encoding a human CFTR polypeptide at the ICP22 locus). In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within one or both loci of the viral ICP4 gene and one or more polynucleotides of the present disclosure within the locus of the viral UL41 gene (for example, a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at one or both ICP4 loci, and a polynucleotide encoding a human CFTR polypeptide at the UL41 locus).In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure within the viral ICP22 gene locus and one or more polynucleotides of the present disclosure within the viral UL41 gene locus (for example, a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide at the ICP22 locus and a polynucleotide encoding a human CFTR polypeptide at the UL41 locus). In some embodiments, a recombinant herpes simplex virus genome comprises... Petition 870260061885, dated 06 / 24 / 2026, page 68 / 151 / 101 one or more polynucleotides of the present disclosure in one or both loci of the viral ICP4 gene, one or more polynucleotides of the present disclosure within the locus of the viral ICP22 gene, and one or more polynucleotides of the present disclosure within the locus of the viral UL41 gene (for example, a recombinant virus carrying a polynucleotide encoding a human CFTR polypeptide in one or both ICP4 loci, a polynucleotide encoding a human CFTR polypeptide in the ICP22 locus, and a polynucleotide encoding a human CFTR polypeptide in the UL41 locus).In some embodiments, a recombinant herpes simplex virus genome comprises one or more polynucleotides of the present disclosure at one or both loci of the viral ICP4 gene, one or more polynucleotides of the present disclosure within the viral ICP22 gene locus, one or more polynucleotides of the present disclosure within the viral UL41 gene locus, one or more polynucleotides of the present disclosure within one or both loci of the viral ICP0 gene, one or more polynucleotides of the present disclosure within the viral ICP27 gene locus, and / or one or more polynucleotides of the present disclosure at the viral ICP47 gene locus.

[0098] In some embodiments, the recombinant herpes virus genome (e.g., a recombinant herpes simplex virus genome) has been engineered to decrease or eliminate the expression of one or more herpes virus genes (e.g., one or more toxic herpes virus genes), such as one or both copies of the HSV ICP0 gene, one or both copies of the HSV ICP4 gene, the HSV ICP22 gene, the HSV UL41 gene, the HSV ICP27 gene, the HSV ICP47 gene, etc. In some embodiments, the recombinant herpes virus genome (e.g., a recombinant herpes simplex virus genome) has been engineered to reduce the cytotoxicity of the recombinant genome (e.g., when introduced into a target cell) compared to a corresponding wild-type herpes virus genome. In some embodiments, the target cell is a Petition 870260061885, dated 06 / 24 / 2026, page 69 / 151 / 101 human cell (primary cells or a cell line derived from them). In some embodiments, the target cell is a mucosal cell. In some embodiments, the target cell is a respiratory tract cell (primary cells or a cell line derived from them). In some embodiments, the target cell is an airway epithelial cell (primary cells or a cell line derived from them). In some embodiments, the target cell is a lung cell (primary cells or a cell line derived from them).In some embodiments, the cytotoxicity of the recombinant herpesvirus genome is reduced by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% compared with a corresponding wild-type herpesvirus genome (e.g., by measuring the relative cytotoxicity of a recombinant ΔKT4 herpes simplex virus genome (one or both copies) versus a wild-type herpes simplex virus genome in a target cell; measurement of the relative cytotoxicity of a recombinant ΔK.T4 (one or both copies) / ΔK herpes simplex virus.T22 genome vs. wild-type herpes simplex virus genome in a target cell, etc.). In some embodiments, the cytotoxicity of the recombinant herpes virus genome is reduced by at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 15 times, at least about. Petition 870260061885, dated 06 / 24 / 2026, p. 70 / 151 / 101 times, at least about 25 times, at least about 50 times, at least about 75 times, at least about 100 times, at least about 250 times, at least about 500 times, at least about 750 times, at least about 1,000 times, or more, compared to a corresponding wild-type herpes simplex virus genome (e.g., measuring the relative cytotoxicity of a recombinant ΔK.T4 herpes simplex virus genome (one or both copies) vs. a wild-type herpes simplex virus genome in a target cell; measuring the relative cytotoxicity of a recombinant ΔK.T4 herpes simplex virus genome (one or both copies) / ΔK.T22 vs.Methods for measuring cytotoxicity are known to those skilled in the art, including, for example, through the use of vital dyes (formazan dyes), protease biomarkers, an MTT assay (or an assay using related tetrazolium salts, such as XTT, MTS, water-soluble tetrazolium salts, etc.), measurement of ATP content, etc.

[0099] In some embodiments, the genome of the recombinant herpes virus (e.g., a recombinant herpes simplex virus genome) has been engineered to reduce its impact on target cell proliferation after exposure of a target cell to the recombinant genome, compared to a corresponding wild-type herpes virus genome. In some embodiments, the target cell is a human cell (primary cells or a cell line derived from them). In some embodiments, the target cell is a mucosal cell. In some embodiments, the target cell is a respiratory tract cell (primary cells or a cell line derived from them). In some embodiments, the target cell is an airway epithelial cell (primary cells or a cell line derived from them). In some embodiments, the target cell is a lung cell (primary cells or a Petition 870260061885, dated 06 / 24 / 2026, page 71 / 151 / 101 (cell line derived from the same). In some embodiments, the proliferation of target cells after exposure to the recombinant genome is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% faster compared with the proliferation of target cells after exposure to a corresponding wild-type herpesvirus genome (e.g., measuring relative cell proliferation after exposure to a herpes simplex virus genome). recombinant ΔK.T4 (one or both copies) versus cell proliferation after exposure to a wild-type herpes simplex virus genome in a target cell; measuring the wild-type herpes simplex virus genome in a target cell; proliferation after exposure to a recombinant ΔKT4 herpes simplex virus genome (one or both copies) / ΔK.T22 versus a wild-type herpes simplex virus genome in a target cell, etc.).In some embodiments, the proliferation of target cells after exposure to the recombinant genome is at least about 1.5 times, at least about 2 times, at least about 3 times, at least about 4 times, at least about 5 times, at least about 6 times, at least about 7 times, at least about 8 times, at least about 9 times, at least about 10 times, at least about 15 times, at least about 20 times, at least about 25 times, at least about 50 times, at least about 75 times, at least about 100 times, at least about 250 times, at least about 500 times, at least about 750 times, or at least about 1,000 times faster. Petition 870260061885, dated 06 / 24 / 2026, page 72 / 151 / 101 comparison with the proliferation of target cells after exposure to a corresponding wild-type herpes simplex virus genome (e.g., measuring relative cell proliferation after exposure to a recombinant herpes simplex virus genome ΔKΊM (one or both copies) vs. cell proliferation after exposure to a wild-type herpes simplex virus genome in a target cell; measuring relative cell proliferation after exposure to a recombinant herpes simplex virus genome ΔK.T4 (one or both copies) OAICP22 vs. a wild-type herpes simplex virus genome in a target cell, etc.). Methods for measuring cell proliferation are known to those skilled in the art, including, for example, through the use of a Ki67 cell proliferation assay, a BrdU cell proliferation assay, etc.

[00100] A vector (e.g., herpes viral vector) may include one or more polynucleotides of the present disclosure in a form suitable for expression of the polynucleotide in a host cell. Vectors may include one or more regulatory sequences operatively linked to the polynucleotide to be expressed (e.g., as described above).

[00101] In some embodiments, a recombinant nucleic acid (e.g., a recombinant herpes simplex virus genome) of the present disclosure comprises one or more of the polynucleotides described herein inserted in any orientation in the recombinant nucleic acid. If the recombinant nucleic acid comprises two or more polynucleotides described herein (e.g., two or more, three or more, etc.), the polynucleotides may be inserted in the same orientation or in orientations opposite to each other. Without wishing to be limited by theory, incorporating two polynucleotides (e.g., two transgenes) into a recombinant nucleic acid (e.g., a vector) in an antisense orientation may help to avoid misreading and ensure proper expression of each polynucleotide. IV. Viruses Petition 870260061885, dated 06 / 24 / 2026, page 73 / 151 / 101

[00102] Certain aspects of the present disclosure relate to viruses comprising any of the recombinant polynucleotides and / or nucleic acids described herein. In some embodiments, the virus is capable of infecting one or more target cells of a subject (e.g., a human). In some embodiments, the virus is suitable for delivering the recombinant polynucleotides and / or nucleic acids to one or more target cells of a subject (e.g., a human). In some embodiments, one or more target cells are human cells. In some embodiments, one or more target cells are one or more CFTR-deficient cells (e.g., one or more cells comprising a genomic mutation in the native CFTR gene). In some embodiments, one or more target cells are one or more mucosal cells. In some embodiments, one or more target cells are one or more airway epithelial cells.In some modalities, one or more target cells are one or more cells of the respiratory tract (e.g., airway epithelial cells (such as goblet cells, ciliated cells, Clara cells, neuroendocrine cells, basal cells, intermediate or parabasal cells, serous cells, brush cells, oncocytes, non-ciliated columnar cells and / or metaplastic cells); alveolar cells (such as type 1 pneumocytes, type 2 pneumocytes and / or non-ciliated cuboidal cells); salivary gland cells in the bronchi (such as serous cells, mucous cells and / or ductal cells); etc.). In some modalities, one or more target cells are one or more lung cells.

[00103] Any suitable virus known in the art may be used, including, for example, adenovirus, adeno-associated virus, retrovirus, lentivirus, sendai virus, herpesvirus, vaccinia virus and / or any hybrid virus or derivative thereof. In some embodiments, the virus is attenuated. In some aspects, the virus is competent for replication. In some embodiments, the virus is defective in replication. In some Petition 870260061885, dated 06 / 24 / 2026, page 74 / 151 / 101 embodiments, the virus has been modified to alter its tissue tropism relative to the tissue tropism of a corresponding unmodified wild-type virus. In some embodiments, the virus has reduced cytotoxicity (e.g., in a target cell) compared to a corresponding wild-type virus. The methods of producing a virus comprising recombinant nucleic acids are well known to those skilled in the art.

[00104] In some embodiments, the virus is a member of the Herpesviridae family of DNA viruses, including, for example, a herpes simplex virus, a varicella-zoster virus, a human cytomegalovirus, a herpesvirus 6A, a herpesvirus 6B, a herpesvirus 7, and a herpesvirus associated with Kaposi's sarcoma, etc. In some embodiments, the herpes virus is attenuated. In some embodiments, the herpes virus is defective in replication. In some aspects, the herpes virus is competent for replication. In some embodiments, the herpes virus has been engineered to reduce or eliminate the expression of one or more herpes virus genes (e.g., one or more toxic herpes virus genes). In some embodiments, the herpes virus has reduced cytotoxicity compared to a corresponding wild-type herpes virus. In some embodiments, the herpes virus is not oncolytic.

[00105] In some embodiments, the herpes virus is a herpes simplex virus. Herpes simplex viruses comprising recombinant nucleic acids can be produced by a process disclosed, for example, in WO2015 / 009952, WO2017 / 176336, WO2019 / 200163, WO2019 / 210219 and / or WO2020 / 006486. In some embodiments, the herpes simplex virus is attenuated. In some embodiments, the herpes simplex virus is defective in replication. In some aspects, the herpes simplex virus is competent for replication. In some embodiments, the herpes simplex virus has been engineered to reduce or eliminate the expression of one or Petition 870260061885, dated 06 / 24 / 2026, p. 75 / 151 / 101 more herpes simplex virus genes (e.g., one or more toxic herpes simplex virus genes). In some embodiments, the herpes simplex virus has reduced cytotoxicity compared to a corresponding wild-type herpes simplex virus. In some embodiments, the herpes simplex virus is not oncolytic. In some embodiments, the herpes simplex virus is an HSV-1 virus, an HSV-2 virus, or any derivative thereof. In some embodiments, the herpes simplex virus is an HSV-1 virus. In some embodiments, the herpes simplex virus is an HSV-1 virus. In some embodiments, the HSV-1 is attenuated. In some embodiments, the HSV-1 is replication-defective. In some embodiments, the HSV-1 is replication-competent. In some embodiments, HSV-1 has been engineered to reduce or eliminate the expression of one or more HSV-1 genes (e.g., one or more toxic HSV-1 genes).In some modalities, HSV-1 reduced cytotoxicity compared to a matching wild-type HSV-1. In some modalities, HSV-1 is not oncolytic.

[00106] In some embodiments, the herpes simplex virus has been modified to alter its tissue tropism relative to the tissue tropism of an unmodified wild-type herpes simplex virus. In some embodiments, the herpes simplex virus comprises a modified envelope. In some embodiments, the modified envelope comprises one or more (e.g., one or more, two or more, three or more, four or more, etc.) mutant herpes simplex virus glycoproteins. Examples of herpes simplex virus glycoproteins may include, but are not limited to, gB, gC, gD, gH, and gL glycoproteins. In some embodiments, the modified envelope alters the tissue tropism of the herpes simplex virus relative to a wild-type herpes simplex virus.

[00107] In some embodiments, the transduction efficiency (in vitro and / or in vivo) of a virus of the present disclosure (for example, a virus of Petition 870260061885, dated 06 / 24 / 2026, p. 76 / 151 / 101 herpes) for one or more target cells (e.g., one or more cells of the respiratory tract) is at least about 25%. For example, the transduction efficiency of the virus to one or more target cells may be at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99%, at least about 99.5% or more. In some embodiments, the virus is a herpes simplex virus and the transduction efficiency of the virus to one or more target cells (e.g., one or more respiratory tract cells) is about 85% to about 100%.In some embodiments, the virus is a herpes simplex virus and the transduction efficiency of the virus to one or more target cells (e.g., one or more cells of the respiratory tract) is at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99% or 100%. Methods for measuring viral transduction efficiency in vitro or in vivo are well known to those skilled in the art, including, for example, qPCR analysis, deep sequencing, western blotting, fluorometric analysis (such as fluorescence in situ hybridization (FISH), fluorescent reporter gene expression, immunofluorescence, FACS), etc. V. Pharmaceutical Compositions and Formulations

[00108] Certain aspects of this disclosure relate to pharmaceutical compositions or formulations comprising any of the recombinant nucleic acids (for example, a herpes virus genome). Petition 870260061885, dated 06 / 24 / 2026, page 77 / 151 / 101 recombinant) and / or virus (for example, a herpes virus comprising a recombinant genome) described herein (such as a herpes simplex virus comprising a recombinant herpes simplex virus genome) and a pharmaceutically acceptable excipient or carrier.

[00109] In some embodiments, the pharmaceutical composition or formulation comprises any one or more of the viruses (e.g., herpes virus) described herein. In some embodiments, the pharmaceutical composition or formulation comprises from about 10⁴ to about 10¹² plaque-forming units (PFU) / mL of the virus. For example, the pharmaceutical composition or formulation may comprise approximately 104 to approximately 1012, approximately 105 to approximately 1012, approximately 106 to approximately 1012, approximately 107 to approximately 1012, approximately 108 to approximately 1012, approximately 109 to approximately 1012, approximately 1010 to approximately 1012, approximately 1011 to approximately 1012, approximately 104 to approximately 1011, approximately 105 to approximately 1011, approximately 106 to approximately 1011, approximately 107 to approximately 1011, approximately 108 to approximately 1011, approximately 109 to approximately 1011, approximately 1010 to approximately 1011, approximately 104 to about 1010, about 105 to about 1010, about 106 to about 1010, about 107 to about 1010, about 108 to about 1010,about 109a about 1010, about 104a about 109, about 105 to about 109, about 106a about 109, about 107a about 109, about 108a about 109, about 104a about 108, about 105a about 108, about 106a about 108, about 107a about 108, about 104a about 107, about 105a about 107, about 106a about 107, about 104a about 106, about 105a about 106, or about 104a about 105PFU / mL of the virus. In some embodiments, the pharmaceutical composition or formulation comprises approximately 10⁴, approximately 10⁵, approximately 10⁶, approximately 10⁷, approximately 10⁸, approximately 10⁹, approximately 10¹⁰, approximately 10¹¹, or approximately 10¹² PFU / mL of the virus.

[00110] Pharmaceutical compositions and formulations can be prepared by mixing the active ingredient(s) (such as an acid). Petition 870260061885, dated 06 / 24 / 2026, page 78 / 151 / 101 recombinant nucleic acid and / or a virus) possessing the desired degree of purity with one or more pharmaceutically acceptable carriers or excipients. Pharmaceutically acceptable carriers or excipients are generally non-toxic to recipients at the dosages and concentrations used and may include, but are not limited to: buffers (such as phosphate, citrate, acetate and other organic acids); antioxidants (such as ascorbic acid and methionine); preservatives (such as octadecyldimethylbenzyl ammonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkyl parabens, catechol, resorcinol, cyclohexanol, 3-pentanol and m-cresol); amino acids (such as glycine, glutamine, asparagine, histidine, arginine, or lysine); low molecular weight (less than about 10 residues) polypeptides; proteins (such as serum albumin, gelatin, or immunoglobulins);Polyols (such as glycerol, for example, formulations including 10% glycerol); hydrophilic polymers (such as polyvinylpyrrolidone); monosaccharides, disaccharides, and other carbohydrates (including glucose, mannose, or dextrins); chelating agents (such as EDTA); sugars (such as sucrose, mannitol, trehalose, or sorbitol); salt-forming counterions (such as sodium); metal complexes (such as Zn-protein complexes); and / or nonionic surfactants (such as polyethylene glycol (PEG)). A complete discussion of pharmaceutically acceptable carriers is available in REMINGTON'S PHARMACEUTICAL SCIENCES (Mack Pub. Co., NJ 1991).

[00111] In some embodiments, the pharmaceutical composition or formulation comprises one or more lipid carriers (e.g., cationic lipid). In some embodiments, the pharmaceutical composition or formulation comprises one or more nanoparticle carriers. Nanoparticles are submicron-sized (less than about 1000 nm) drug delivery vehicles that can carry encapsulated drugs (such as small synthetic molecules, proteins, peptides, Petition 870260061885, dated 06 / 24 / 2026, p. 79 / 151 / 101 cells, viruses and nucleic acid-based biotherapeutics) for rapid or controlled release. A variety of molecules (e.g., proteins, peptides, recombinant nucleic acids, etc.) can be efficiently encapsulated in nanoparticles using processes well known in the art. In some embodiments, a molecule encapsulated in a nanoparticle may refer to a molecule (such as a virus) that is contained within the nanoparticle or bound to and / or associated with the surface of the nanoparticle, or any combination thereof. Nanoparticles for use in the compositions or formulations described herein may be any type of biocompatible nanoparticle known in the art, including, for example, nanoparticles comprising poly(lactic acid), poly(glycolic acid), PLGA, PLA, PGA and any combinations thereof (see, for example, Vauthier et al. Adv Drug Del Rev.(2003) 55: 519-48; US2007 / 0148074; US2007 / 0092575; US2006 / 0246139; US5753234; US7081483; e. WO2006 / 052285).

[00112] In some embodiments, the pharmaceutically acceptable carrier or excipient may be adapted or suitable for any route of administration known in the art, including, for example, intravenous, intramuscular, subcutaneous, cutaneous, oral, intranasal, intratracheal, sublingual, buccal, topical, transdermal, intradermal, intraperitoneal, intraorbital, intravitreal, subretinal, transmucosal, intra-articular, by implantation, by inhalation, intrathecal, intraventricular and / or intranasal administration. In some embodiments, the pharmaceutically acceptable carrier or excipient is adapted or suitable for oral, intranasal, intratracheal and / or inhalation administration. In some embodiments, the pharmaceutically acceptable carrier or excipient is adapted or suitable for inhalation administration. In some embodiments, the pharmaceutically acceptable carrier or excipient is adapted or suitable for non-invasive inhalation administration. In some embodiments, Petition 870260061885, dated 06 / 24 / 2026, page 80 / 151 / 101 the carrier or pharmaceutically acceptable excipient is adapted or suitable for nebulization (e.g., using a vibrating mesh nebulizer).

[00113] In some embodiments, the pharmaceutical composition or formulation is adapted or suitable for any route of administration known in the art, including, for example, intravenous, intramuscular, subcutaneous, cutaneous, oral, intranasal, intratracheal, sublingual, buccal, topical, transdermal, intradermal, intraperitoneal, intraorbital, intravitreal, subretinal, transmucosal, intra-articular, by implantation, by inhalation, intrathecal, intraventricular or intranasal administration. In some embodiments, the pharmaceutical composition or formulation is adapted or suitable for oral, intranasal, intratracheal or inhalation administration. In some embodiments, the pharmaceutical composition or formulation is adapted or suitable for inhalation administration. In some embodiments, the pharmaceutical composition or formulation is adapted or suitable for non-invasive inhalation administration.In some embodiments, the pharmaceutical composition or formulation is adapted or made suitable for nebulization (for example, using a vibrating mesh nebulizer).

[00114] In some embodiments, the pharmaceutical composition or formulation further comprises one or more additional components. Examples of additional components may include, but are not limited to, binding agents (e.g., pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose, etc.); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethylcellulose, polyacrylates or calcium hydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc, silica, colloidal silicon dioxide, stearic acid, metallic stearates, hydrogenated vegetable oils, corn starch, polyethylene glycols, sodium benzoate, sodium acetate, etc.); disintegrants (e.g., starch, starch glycolate). Petition 870260061885, dated 06 / 24 / 2026, page 81 / 151 / 101 sodium, etc.); humectants (e.g., sodium lauryl sulfate, etc.); saline solutions; alcohols; polyethylene glycols; gelatin; lactose; amylase; magnesium stearate; talc; silicic acid; viscous paraffin; hydroxymethylcellulose; polyvinylpyrrolidone; sweeteners; flavorings; perfumes; colorants; moisturizers; sunscreens; antibacterial agents; agents capable of stabilizing polynucleotides or preventing their degradation and the like. In some embodiments, the pharmaceutical composition or formulation comprises a phosphate buffer. In some embodiments, the pharmaceutical composition or formulation comprises glycerol (for example, in about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, etc.). In some embodiments, the pharmaceutical composition or formulation comprises a phosphate buffer and glycerol.In some embodiments, the pharmaceutical composition or formulation comprises less than about 15%, less than about 14%, less than about 13%, less than about 12%, less than about 11%, less than about 10%, less than about 9%, less than about 8%, less than about 7%, less than about 6%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, less than about 1%, less than about 0.5%, or less than about 0.1% of glycerol. In some embodiments, the pharmaceutical composition or formulation does not comprise glycerol.

[00115] Pharmaceutical compositions and formulations to be used for in vivo administration are generally sterile. Sterility can be easily achieved, for example, by filtration through sterile filtration membranes.

[00116] In some embodiments, any of the recombinant nucleic acids, viruses and / or pharmaceutical compositions or formulations described herein may be used to deliver one or more polynucleotides encoding a CFTR polypeptide into one or more cells of a subject (by Petition 870260061885, dated 06 / 24 / 2026, p. 82 / 151 / 101 (e.g., one or more CFTR-deficient cells, one or more cells harboring a CFTR gene mutation, one or more respiratory tract cells, etc.). In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations described herein may be used in the treatment of a disease or condition that would benefit from the expression of a CFTR polypeptide (e.g., a disease associated with a CFTR deficiency and / or a disease associated with a CFTR gene mutation). In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations described herein may be used in the prevention or treatment of a chronic lung disease (such as cystic fibrosis, COPD, etc.).In some embodiments, any of the recombinant nucleic acids, viruses, and / or pharmaceutical compositions or formulations described herein may be used in the prevention or treatment of cystic fibrosis.

[00117] In some embodiments, any of the recombinant nucleic acids, viruses and / or pharmaceutical compositions or formulations described herein may be used in the preparation of a medicament useful for delivering one or more polynucleotides encoding a CFTR polypeptide into one or more cells of a subject (e.g., one or more CFTR-deficient cells, one or more cells harboring a CFTR gene mutation, one or more respiratory tract cells, etc.). In some embodiments, any of the recombinant nucleic acids, viruses and / or pharmaceutical compositions or formulations described herein may be used in the preparation of a medicament useful for the prevention or treatment of a disease or condition that would benefit from the expression of a CFTR polypeptide (e.g., a disease associated with a CFTR deficiency and / or a disease associated with a CFTR gene mutation).In some embodiments, any of the recombinant nucleic acids, viruses and / or pharmaceutical compositions or formulations described herein may be used. Petition 870260061885, dated 06 / 24 / 2026, pp. 83 / 151 / 101, concerning the preparation of a medicament useful for the prevention or treatment of a chronic lung disease (such as cystic fibrosis, COPD, etc.). In some embodiments, any of the recombinant nucleic acids, viruses and / or pharmaceutical compositions or formulations described herein may be used in the preparation of a medicament useful for the prevention or treatment of cystic fibrosis. VI. Methods

[00118] Certain aspects of the present disclosure relate to enhancing, increasing, raising and / or supplementing the levels of a CFTR polypeptide in one or more cells of a subject, including administering to the subject any of the recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. In some embodiments, the subject's genome comprises a mutation (e.g., a loss-of-function mutation) in an endogenous CFTR gene (one or both copies). In some embodiments, the subject suffers from a chronic lung disease, e.g., cystic fibrosis, COPD, etc. In some embodiments, the subject suffers from cystic fibrosis.

[00119] In some embodiments, administration of recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation to the subject increases CFTR levels (transcription or protein levels) by at least about 2-fold in one or more contacted or treated cells of the subject, compared to endogenous CFTR levels in one or more corresponding untreated cells in the subject. For example, administration of recombinant nucleic acid, virus, drug, and / or pharmaceutical composition or formulation may increase CFTR levels (transcription or protein levels) by at least about 2-fold, at least about 3-fold, at least about 4-fold, at least about 5-fold, at least about 6-fold, at least about 7-fold, at least Petition 870260061885, dated 06 / 24 / 2026, page 84 / 151 / 101 approximately 8 times, at least approximately 9 times, at least approximately 10 times, at least approximately 15 times, at least approximately 20 times, at least approximately 25 times, at least approximately 50 times, at least approximately 75 times, at least approximately 100 times, at least approximately 250 times, at least approximately 500 times, at least approximately 750 times, at least approximately 1000 times, or more in one or more contacted or treated cells of the subject, compared to endogenous CFTR levels in one or more corresponding untreated cells in the subject. In some modalities, one or more cells contacted or treated are one or more cells of the respiratory tract (e.g., one or more cells of the airway epithelium and / or one or more cells of the submucosal glands).The methods for measuring transcription or protein levels in a sample are well known to those skilled in the art, including, for example, qPCR, western blot, mass spectrometry, etc.

[00120] Other aspects of the present disclosure relate to a method of reducing cellular sodium levels in a subject in need thereof, comprising administering to the subject any of the recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. In some embodiments, the subject's genome comprises a mutation (e.g., a loss-of-function mutation) in an endogenous CFTR gene (one or both copies). In some embodiments, the subject suffers from a chronic lung disease, e.g., cystic fibrosis, COPD, etc. In some embodiments, the subject suffers from cystic fibrosis.

[00121] In some modalities, the administration of recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation to the subject decreases intracellular sodium levels by at least about 10%, at least about 15%, at least about 20%, by Petition 870260061885, dated 06 / 24 / 2026, page 85 / 151 / 101 less about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 99% or more in one or more contacted or treated cells, compared with intracellular sodium levels in one or more corresponding untreated cells in the subject. The methods of measuring intracellular sodium levels are generally known to those skilled in the art.

[00122] Other aspects of the present disclosure relate to a method for improving a measure of at least one respiratory volume in a subject in need thereof, comprising administering to the subject any of the recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. In some embodiments, the subject's genome comprises a mutation (e.g., a loss-of-function mutation) in an endogenous CFTR gene (one or both copies). In some embodiments, the subject suffers from a chronic lung disease, e.g., cystic fibrosis, COPD, etc.

[00123] In some embodiments, administration of recombinant nucleic acid, virus, drug and / or pharmaceutical composition or formulation to the subject improves a measure of at least one respiratory volume by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least Petition 870260061885, dated 06 / 24 / 2026, p. 86 / 151 / 101 less about 85%, at least about 90%, at least about 95%, at least about 99% or more compared to at least one reference respiratory volume measured in the subject before treatment. Examples of adequate respiratory volumes that can be measured include, for example: Total lung capacity (TLC), the volume in the lungs at maximum inflation; Tidal volume (TV), the volume of air that enters or leaves the lungs during quiet breathing; Residual volume (RV), the volume of air remaining in the lungs after a maximal expiration; Expiratory reserve volume (ERV), the maximum volume of air that can be expired (above tidal volume) during a forced expiration; Inspiratory reserve volume (ERV), the maximum volume of air that can be inhaled in the end-inspiratory position; Inspiratory capacity (IC), the sum of IRV and TV;Vital inspiratory capacity (VIC), the maximum volume of air inspired from the point of maximum expiration; Vital Capacity (VC), the volume of air inspired after the deepest inspiration; Functional residual capacity (FRC), the volume of lungs in the end-expiratory position; Forced vital capacity (FVC), the determination of vital capacity during a forced maximal expiratory effort; Forced Expiratory Volume (time) (FEVt), the volume of air expired under forced conditions in the first t seconds; Forced Inspiratory Flow (FIF), a specific measure of the forced inspiratory curve; Peak Expiratory Flow (PEF), the highest forced expiratory flow measured with a peak flow meter; Maximum voluntary ventilation (MVV), the volume of air expired in a specific period during repetitive maximal effort; etc. Methods of measuring respiratory volumes are generally known to those skilled in the art.

[00124] Other aspects of the present disclosure relate to a method of reducing or preventing chronic bacterial infections of the lungs of a subject in need thereof, comprising administering to the subject any of the recombinant nucleic acids, viruses, Petition 870260061885, dated 06 / 24 / 2026, page 87 / 151 / 101 medicines and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. In some embodiments, the subject's genome comprises a mutation (e.g., a loss-of-function mutation) in an endogenous CFTR gene (one or both copies). In some embodiments, the subject suffers from a chronic lung disease, e.g., cystic fibrosis, COPD, etc. In some embodiments, the subject suffers from cystic fibrosis. Direct and indirect methods of monitoring bacterial infections in the lungs, including improvements thereof, are known to those skilled in the art, including, for example, performing: blood tests or cultures, oximetry, arterial blood gas measurements, bronchoscopy, transtracheal mucus cultures, lung biopsies, thoracentesis, computed tomography scans, etc.

[00125] Other aspects of the present disclosure relate to a method of reducing, preventing, or treating chronic lung inflammation in a subject in need thereof, comprising administering to the subject any of the recombinant nucleic acids, viruses, medications, and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. In some embodiments, the subject's genome comprises a mutation (e.g., a loss-of-function mutation) in an endogenous CFTR gene (one or both copies). In some embodiments, the subject suffers from a chronic lung disease, e.g., cystic fibrosis, COPD, etc. In some embodiments, the subject suffers from cystic fibrosis. Methods of measuring lung inflammation, including improvements thereof, are well known to those skilled in the art, including, for example, measuring exhaled nitric oxide, determining the percentage of eosinophils in sputum and / or blood, etc.

[00126] Other aspects of the present disclosure relate to a method of reducing, inhibiting or treating the progressive destruction of the lung in a subject in need, comprising administering to the subject Petition 870260061885, dated 06 / 24 / 2026, page 88 / 151 / 101 any of the recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. In some embodiments, the subject's genome comprises a mutation (e.g., a loss-of-function mutation) in an endogenous CFTR gene (one or both copies). In some embodiments, the subject suffers from a chronic lung disease, e.g., cystic fibrosis, COPD, etc. In some embodiments, the subject suffers from cystic fibrosis. The methods of measuring lung destruction are well known to those skilled in the art, including, for example, the methods described by Saetta et al. (Am Rev Respir Dis. 1985 May;131(5):764-9).

[00127] Other aspects of the present disclosure relate to a method for providing prophylactic, palliative or therapeutic relief of one or more signs or symptoms of cystic fibrosis in a subject in need thereof, comprising administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. In some embodiments, the subject's genome comprises a mutation (e.g., a loss-of-function mutation) in an endogenous CFTR gene (one or both copies).

[00128] The signs and symptoms of cystic fibrosis may include, but are not limited to: persistent cough producing thick mucus; thick, sticky mucus that accumulates in the airways; wheezing; shortness of breath; sinusitis; repeated lung infections; inflamed nasal passages; bronchiectasis; nasal polyps; hemoptysis; pneumothorax; pancreatitis; recurrent pneumonia; respiratory arrest; and any combination thereof.

[00129] Other aspects of the present disclosure relate to a method for providing prophylactic, palliative or therapeutic relief to one or more Petition 870260061885, dated 06 / 24 / 2026, page 89 / 151 / 101 signs or symptoms of COPD in a subject in need thereof, comprising administering to the subject an effective amount of any of the recombinant nucleic acids, viruses, medications and / or pharmaceutical compositions or formulations described herein. In some embodiments, the subject is a human. In some embodiments, the subject is a smoker or ex-smoker.

[00130] The signs and symptoms of COPD may include, but are not limited to: shortness of breath; wheezing; chest tightness; excess mucus in the lungs; a chronic cough; cyanosis; frequent respiratory infections; and any combination thereof.

[00131] The recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations described herein may be administered by any suitable method or route known in the art, including, without limitation, oral, intranasal, intratracheal, sublingual, buccal, topical, rectal, inhalation, transdermal, subcutaneous, intradermal, intravenous, intra-arterial, intramuscular, intracardiac, intraosseous, intraperitoneal, transmucosal, vaginal, intravitreal, intraorbital, subretinal, intra-articular, periarticular, local and / or epicutaneous routes or any combination thereof. This disclosure therefore covers methods of delivering any of the recombinant nucleic acids, viruses, drugs or pharmaceutical compositions or formulations described herein to an individual (e.g., an individual with, or at risk of developing, a chronic lung disease such as cystic fibrosis).

[00132] In some embodiments, recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations described herein are administered orally, intranasally, intratracheally and / or by inhalation. Methods of delivering drugs to the lungs by oral, intranasal, intratracheal and / or inhaled routes of administration are generally known to one skilled in the art (see, for example, Gardenhire et al. A Petition 870260061885, dated 06 / 24 / 2026, pp. 90 / 151 / 101 Guide to Aerosol Delivery Devices for Respiratory Therapists, 4th Edition, American Association for Respiratory Care, 2017; Patil et al. Pulmonary drug delivery strategies: a concise, systematic review, Lung India. 2012. 29(1):44-9; Marx et al. Intranasal Drug Administration - An Attractive Delivery Route for Some Drugs, 2015).

[00133] In some embodiments, recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations are delivered to the lungs by inhalation of an aerosol formulation. Inhalation may occur through the nose and / or mouth of the subject. Exemplary devices for delivering recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations to the lung may include, without limitation, dry powder inhalers, pressurized metered-dose inhalers, soft mist inhalers, nebulizers (e.g., jet nebulizers, ultrasonic nebulizers, vibrating mesh nebulizers), collision jets, extruded jets, surface wave microfluidic atomization, capillary aerosol generation, electrohydrodynamic aerosol devices, etc. (see, for example, Carvalho and McConville. The function and performance of aqueous devices for halation therapy.(2016) Journal of Pharmacy and Pharmacology.

[00134] Liquid formulations can be administered to a subject's lungs, for example, using a pressurized metered-dose inhaler (pMDI). pMDIs generally include at least two components: a canister in which the liquid formulation is held under pressure in combination with one or more propellants, and a receptacle used to retain and actuate the canister. The canister may contain a single dose or multiple doses of the formulation. The canister may include a valve, typically a metering valve, from which the contents of the canister can be discharged. The aerosolized drug is dispensed from the pMDI by applying force to the canister to push it toward the receptacle, thereby opening the valve and Petition 870260061885, dated 06 / 24 / 2026, pp. 91 / 151 / 101, causing the drug particles to be transported from the valve through the receptacle outlet. After exiting the container, the liquid formulation is atomized, forming an aerosol. pMDIs typically employ one or more propellants to pressurize the contents of the container and to propel the liquid formulation out of the receptacle outlet, forming an aerosol. Any suitable propellants may be used and may take a variety of forms, including, for example, a compressed gas or a liquefied gas.

[00135] Liquid formulations can be administered to a subject's lungs, for example, using a nebulizer. Nebulizers are liquid aerosol generators that convert the liquid formulation into mists or clouds of small droplets, often with diameters less than about 5 microns of mass median aerodynamic diameter, which can be inhaled into the lower respiratory tract. The droplets carry the active agent(s) to the nose, upper respiratory tract, and / or deep lungs when the aerosol cloud is inhaled. Any type of nebulizer known in the art can be used to administer the formulation to a patient, including, without limitation, pneumatic (jet) nebulizers, electromechanical nebulizers (e.g., ultrasonic nebulizers, vibrating mesh nebulizers, etc.), etc. Pneumatic (jet) nebulizers use a pressurized gas supply as the driving force for atomizing the liquid formulation.Compressed gas is supplied through a nozzle or jet to create a low-pressure field that draws in a surrounding liquid formulation and cuts it into a thin film or filaments. The film or filaments are unstable and break down into small droplets that are carried by the compressed gas stream to the inspiratory breath. Deflectors inserted into the droplet plume screen remove the larger droplets and return them to the bulk liquid reservoir. Electromechanical nebulizers use mechanical force. Petition 870260061885, dated 06 / 24 / 2026, pp. 92 / 151 / 101, electrically generated to atomize liquid formulations. The electromechanical driving force can be applied, for example, by vibrating the liquid formulation at ultrasonic frequencies or by forcing the bulk liquid through small orifices into a thin film. The forces generate thin liquid films or filament streams that separate into small droplets to form a slow-moving aerosol stream, which can be carried in an inspiratory flow. In some embodiments, the nebulizer is a vibrating mesh nebulizer. Examples of vibrating mesh nebulizers include, for example, the Phillips InnoSpire, the Aerogen Solo, the PARI eFlow, etc.

[00136] Liquid formulations can be administered to a subject's lungs, for example, using an electrohydrodynamic (EHD) aerosol device. EHD aerosol devices use electrical energy to aerosolize liquid drug solutions or suspensions.

[00137] Dry powder formulations can be administered to a subject's lungs, for example, using a dry powder inhaler (DPI). DPIs typically use a mechanism such as a gas explosion to create a cloud of dry dust within a canister, which can then be inhaled by the subject. In a DPI, the dose to be administered is stored in the form of an unpressurized dry powder, and upon actuation of the inhaler, the powder particles are inhaled by the subject. In some cases, a compressed gas may be used to dispense the powder, similar to pMDIs. In some cases, the DPI may be breath-triggered (an aerosol is created in precise response to inspiration). Typically, dry powder inhalers administer a dose of less than a few tens of milligrams per inhalation to avoid provoking coughing.Examples of DPIs include, for example, the Turbohaler® inhaler (AstraZeneca), the Clickhaler® inhaler (Innovata), the Diskus® inhaler (Glaxo), the EasyHaler® (Orion), the Exubera® inhaler (Pfizer), etc. Petition 870260061885, dated 06 / 24 / 2026, p. 93 / 151 / 101

[00138] In some embodiments, recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations are administered once to the subject. In some embodiments, recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions are administered at least twice (e.g., at least 2 times, at least 3 times, at least 4 times, at least 5 times, at least 10 times, etc.) to the subject.In some modalities, at least about 1 hour (for example, at least about 1 hour, at least about 6 hours, at least about 12 hours, at least about 18 hours, at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 15 days, at least about 20 days, at least about 30 days, at least about 40 days, at least about 50 days, at least about 60 days, at least about 70 days, at least about 80 days, at least about 90 days, at least about 100 days, at least about 120 days, etc.) passes between administrations (for example, between the first and second administrations, between the second and third administrations, etc.).In some embodiments, recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations are administered to the subject one, two, three, four, five, or more times per day. In some embodiments, recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations are administered to the subject one, two, three, four, five, or more times per month. In some embodiments, recombinant nucleic acids, viruses, drugs, and / or pharmaceutical compositions or formulations are administered to the subject one, two, three, four, five, or more times per year. VII. Host Cells

[00139] Certain aspects of this disclosure relate to a or Petition 870260061885, dated 06 / 24 / 2026, p. 94 / 151 / 101 plus host cells comprising any of the recombinant nucleic acids described herein. Any suitable host cell (prokaryotic or eukaryotic) known in the art may be used, including, for example: prokaryotic cells including eubacteria, such as Gram-negative or Gram-positive organisms, for example Enterobacteriaceae, such as Escherichia (e.g., E. coli), Enterobacter, Erminia, Klebsiella, Proteus, Salmonella (e.g., S. typhimurium), Serratia (e.g., S. marcescans), and Shigella, as well as Bacilli such as B. subtilis and B. licheniformis; fungal cells (e.g., S. cerevisiae); insect cells (e.g., S2 cells, etc.).); and mammalian cells, including SV40-transformed monkey kidney cell line CV1 (COS-7, ATCC CRL 1651), human embryonic kidney cell line (293 or 293 subcloned cells for growth in suspension culture), baby hamster kidney cells (BHK, ATCC CCL 10), mouse Sertoli cells (TM4), monkey kidney cells (CV1 ATCC CCL 70), African green monkey kidney cells (VERO-76, ATCC CRL-1587), human cervical carcinoma cells (HELA, ATCC CCL 2), canine kidney cells (MDCK, ATCC CCL 34), buffalo rat liver cells (BRL 3A, ATCC CRL 1442), human lung cells (W138, ATCC CCL 75), human liver cells (Hep G2, HB 8065), mouse mammary tumor (MMT). 060562, ATCC CCL51), TRI cells, MRC 5 cells, FS4 cells, human hepatoma cell line (Hep G2), Chinese hamster ovary (CHO) cells, including DHFR CHO cells, and myeloma cell lines such as NS0 and Sp2 / 0.In some embodiments, the host cell is a human or non-human primate cell. In some embodiments, the host cells are cells of a cell line. Examples of suitable host cells or cell lines may include, but are not limited to, 293, HeLa, SH-Sy5y, Hep G2, CACO-2, A549, L929, 3T3, K562, CHO-K1, MDCK, HUVEC, Vero, N20 cells, COS-7, PSN1, VCaP, CHO and. Petition 870260061885, dated 06 / 24 / 2026, page 95 / 151 / 101 similar.

[00140] In some embodiments, the recombinant nucleic acid is a herpes simplex viral vector. In some embodiments, the recombinant nucleic acid is a herpes simplex virus amplicon. In some embodiments, the recombinant nucleic acid is an HSV-1 amplicon or HSV-1 hybrid amplicon. In some embodiments, a host cell comprising a helper virus is contacted with an HSV1 amplicon or HSV-1 hybrid amplicon described herein, resulting in the production of a virus comprising one or more recombinant nucleic acids described herein. In some embodiments, the virus is collected from the supernatant of the contacted host cell. Methods of generating viruses by contacting host cells comprising a helper virus with an HSV-1 amplicon or HSV-1 hybrid amplicon are known in the art.

[00141] In some embodiments, the host cell is a yeast cell. In some embodiments, the complementary host cell expresses one or more genes that are inactivated in any of the viral vectors described herein. In some embodiments, the complementary host cell is brought into contact with a recombinant herpes virus genome (e.g., a recombinant herpes simplex virus genome) described herein. In some embodiments, contact of a complementary host cell with a recombinant herpes virus genome results in the production of a herpes virus comprising one or more recombinant nucleic acids described herein. In some embodiments, the virus is collected from the supernatant of the contacted host cell. Methods for generating viruses by contacting complementary host cells with a recombinant herpes simplex virus are generally described in WO2015 / 009952, WO2017 / 176336, WO2019 / 200163, WO2019 / 210219 and / or WO2020 / 006486. VIII. Manufacturing Items or Kits Petition 870260061885, dated 06 / 24 / 2026, p. 96 / 151 / 101

[00142] Certain aspects of this disclosure relate to an article of manufacture or a kit comprising any of the recombinant nucleic acids, viruses, medicaments and / or pharmaceutical compositions or formulations described herein. In some embodiments, the article of manufacture or kit comprises a package insert containing instructions for administering the recombinant nucleic acid, virus, medicament and / or pharmaceutical composition or formulation to treat a CFTR deficiency (e.g., in a subject carrying homozygous loss-of-function CFTR genetic mutations) and / or to provide prophylactic, palliative or therapeutic relief of one or more signs or symptoms of a chronic lung disease (such as cystic fibrosis or COPD). In some embodiments, the article of manufacture or kit further comprises a device for administering (e.g., aerosolizing) the recombinant nucleic acid, virus, medicament and / or pharmaceutical composition or formulation.In some forms, the device is a nebulizer (for example, a vibrating mesh nebulizer).

[00143] Suitable containers for recombinant nucleic acids, viruses, drugs and / or pharmaceutical compositions or formulations may include, for example, bottles, vials, bags, tubes and syringes. The container may be formed from a variety of materials, such as glass, plastic (such as polyvinyl chloride or polyolefin) or metal alloy (such as stainless steel or hastelloy). In some embodiments, the container comprises a label on or associated with the container, wherein the label indicates instructions for use. The article of manufacture or kit may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, inhalers, nebulizers, intranasal administration devices, a package insert and the like.

[00144] The descriptive report is considered sufficient to allow Petition 870260061885, dated 06 / 24 / 2026, p. 97 / 151 84 / 101 who are skilled in the art and practice the present disclosure. Several modifications of the invention in addition to those presented and described herein will be apparent to those skilled in the art from the preceding description and are within the scope of the appended claims. EXAMPLES

[00145] The invention will be more fully understood by reference to the following examples. They should not, however, be construed as limiting the scope of the invention. It is understood that the examples and embodiments described in this document are for illustrative purposes only, and that various modifications or alterations in light thereof will be suggested for persons skilled in the art and should be included within the spirit and scope of this application and the scope of the appended claims. Example 1: Modified herpes simplex virus vectors encoding a human CFTR protein.

[00146] To make modified herpes simplex virus genome vectors capable of expressing CFTR polypeptides in a target mammalian cell (such as lung cells), a herpes simplex virus genome (FIG. IA) is first modified to inactivate one or more herpes simplex virus genes. These modifications can decrease the genome's toxicity in mammalian cells. Then, variants of these modified / attenuated recombinant viral constructs are generated so that they carry one or more polynucleotides encoding the desired CFTR polypeptide. These variants include: 1) an HSV-1 genome modified with recombinant AICP4 comprising expression cassettes containing the coding sequence (e.g., SEQ ID NO: 2) of a human CFTR polypeptide (e.g., SEQ ID NO: 5) under the control of a heterologous promoter integrated into each ICP4 locus (FIG.IB); 2) an HSV1 genome modified with recombinant AICP4 / AUL41 comprising expression cassettes containing the coding sequence of a CFTR polypeptide. Petition 870260061885, dated 06 / 24 / 2026, page 98 / 151 / 101 human under the control of a heterologous promoter integrated into each ICP4 locus (FIG. 1C); 3) an HSV-1 genome modified with recombinant Δ!ΟΓ4 / ΔHΡ41 comprising an expression cassette containing the coding sequence of a human CFTR polypeptide under the control of a heterologous promoter integrated into the UL41 locus (FIG. 1D); 4) an HSV-1 genome modified with recombinant ΔK.T4 / ΔKΓ22 comprising expression cassettes containing the coding sequence of a human CFTR polypeptide under the control of a heterologous promoter integrated into each ICP4 locus (FIG. 1E); 5) an HSV-1 genome modified with recombinant ΔK.T4 / MCP22 comprising an expression cassette containing the coding sequence of a human CFTR polypeptide under the control of a heterologous promoter integrated into the ICP22 locus (FIG. 1F); 6) an HSV-1 genome modified with ΔK.T4 / ΔHP41 / ΔK.7) a recombinant ΔK.T4 / ΔHP41 / ΔK.T22 HSV-1 genome comprising an expression cassette containing the coding sequence of a human CFTR polypeptide under the control of a heterologous promoter integrated into each ICP4 locus (FIG. 1G); and 8) a recombinant ΔK.T4 / ΔHP41 / ΔK.T22 HSV-1 genome comprising an expression cassette containing the coding sequence of a human CFTR polypeptide under the control of a heterologous promoter integrated into the UL41 locus (FIG. 1H); and 9) a recombinant ΔK.T4 / ΔHP41 / ΔK.T22 HSV-1 genome comprising an expression cassette containing the coding sequence of a human CFTR polypeptide under the control of a heterologous promoter integrated into the ICP22 locus (FIG. 1I).

[00147] These modified herpes simplex virus genome vectors are transfected into cells modified to express one or more herpes virus genes. These modified cells secrete into the cell culture supernatant a replicating defective herpes simplex virus with the modified genomes packaged within it. The supernatant is Petition 870260061885, dated 06 / 24 / 2026, page 99 / 151 / 101 then collected, concentrated and sterilized by filtration through a 5 μm filter. Example 2: In vitro construction and characterization of an HSV-1 vector encoding human CFTR in 2D cultures.

[00148] Initial clinical trials of lung gene therapy took place in the early 1990s, following the discovery of the genetic defect responsible for cystic fibrosis. Recombinant adenovirus was one of the first vectors tested for CFTR delivery; however, adeno-based vectors failed in these trials primarily due to the scarcity of viral receptors on the apical surface of the lung and the severity of the host immune response to repeated viral delivery. Other viral gene therapy vectors administered to CF patients were based on adeno-associated viruses (various AAV serotypes were tested in the CF clinical setting). Large repeated-delivery studies of AAV-based gene therapy vectors yielded disappointing results in improving CF lung function in dosed patients.Much like adenovirus, recombinant AAV vectors do not effectively infect and form the apical surface of the lung, and due to physical limitations of the encoded payload size, AAV vectors do not effectively deliver full-length human CFTR. Despite more than two decades of intensive effort, virus-based gene therapies have yet to help patients with CF (or any other obstructive pulmonary disease).

[00149] Currently, according to the US Cystic Fibrosis Foundation, there are no ongoing clinical trials of viral gene therapies in CF, and only two virus-based gene therapy vectors are in preclinical development (both based on AAV, a vector that, as noted above, has already failed in several clinical trials in CF patients). Instead, the focus has shifted from virus-based vectors to non-viral methods of CFTR delivery (e.g., DNA plasmids or mRNAs). Petition 870260061885, dated 06 / 24 / 2026, pp. 100 / 151 / 101 complexed with liposomes). Unfortunately, these nonviral vectors have had only limited success, due, at least in part, to the significant obstacles faced by product instability and / or inefficient distribution / transfection of liposomal formulations. In total, more than 25 clinical trials involving over 470 patients testing viral and nonviral gene vectors have shown no clinical benefit, largely due to inefficient gene transfer to target cells and immune-mediated host clearance after repeated exposure.

[00150] To this end, a herpes simplex virus type 1 (HSV1) vector encoding full-length human CFTR (HSV-CFTR) has been developed as a novel gene therapy for the treatment of CF patients. Without wishing to be limited by theory, an HSV-based approach is believed to overcome many of the obstacles experienced by other gene therapy vectors for CF, including the ability to encode full-length human CFTR, high target cell transduction efficiency (HSV preferentially infects the apical membrane of polarized epithelial cells), viral stability, and the established clinical safety of repeated administration of a product employing the same HSV-CFTR viral backbone in the context of the highly inflammatory environment of wounded skin (ClinicalTrials.gov identifier: NCT03536143).The following example describes experiments showing that this novel HSV-based gene therapy vector was able to express functional full-length human CFTR in cystic fibrosis patient-derived airway epithelial cells (SAECs) in a dose-dependent manner.

[00151] HSV-CFTR was constructed as described in Example 1 above. Primary SAECs from CF patients cultured in 2D culture were either uninfected (sham) or infected with HSV-CFTR at multiplicities of infection (MOIs) of 0, 3, 1, or 3. Human CFTR expression was assessed 48 hours post-infection in cells harvested by PCR. Petition 870260061885, dated 06 / 24 / 2026, page 101 / 151 / 101 quantitative reverse transcription (qRT-PCR). Codon-optimized CFTR transcripts were detected in primary infected CF SAECs at an MOI as low as 0.3 and appeared to show a dose-dependent increase in transgene expression up to an MOI of 3.0 (FIG. 2). Little or no exogenous CFTR RNA was observed in sham-infected control samples, demonstrating the assay's specificity for the HSV-encoded human transgene.

[00152] CFTR protein expression in primary CF SAECs infected with HSV-CFTR was assessed by Western blot analysis. GAPDH was used as a control to ensure consistent loading of samples. CF patient SAECs overexpressed human CFTR when infected with HSV-CFTR, compared to control cells with sham infection (FIG. 3). Interestingly, while the endogenous CFTR protein in sham-infected cells resolved as a single band slightly larger than 150 kDa (the predicted size of full-length human CFTR is 168 kDa), the exogenous CFTR protein expressed in HSV-CFTR transduced cells appeared as a significantly larger doublet. Human CFTR is known to exist in three different forms depending on the glycosylation state: (1) non-glycosylated; (2) glycosylated core; and (3) fully mature glycosylated complex (Scanlin, 2001, Respir Res, 2(5), pp. 276-9).The appearance of a single low molecular weight band in cells from a sham CF-infected patient suggested that the endogenous (mutant) protein exists only in the non-glycosylated form, indicative of an immature protein variant that does not adequately traffick through the endoplasmic reticulum (ER) to the cell surface. In stark contrast, the appearance of the two larger forms of CFTR in HSV-CFTR-infected cells revealed extensive post-translational modification of the human transgene, likely representing the glycosylated core and complex glycosylated variants. Petition 870260061885, dated 06 / 24 / 2026, page 102 / 151 / 101 CFTR, suggesting proper maturation and trafficking of the exogenous protein through the ER.

[00153] CFTR protein expression and relative localization were subsequently examined by immunofluorescence. Primary SAECs from CF patients were transduced with HSV-CFTR at the indicated MOIs for 48 hours, and immunofluorescence staining for human CFTR was performed. A sham infection control sample was added to show baseline levels and cellular localization of the endogenous mutant CFTR protein in these diseased cells. When analyzed in the context of control cells, immunofluorescence data demonstrated that transduced SAECs exhibited a dose-dependent increase in HSVC-CFTR expression of the CFTR protein (FIG. 4A). When comparing the relative cellular localization of CFTR expressed in SAECs from CF patients infected with HSV-CFTR and sham infection (FIG.4B), CFTR expressed in uninfected cells appeared to be relegated to the perinuclear region (suggestive of trapping and turnover in the ER), while CFTR was found throughout the cytoplasm and on the cell surface of transduced HSVCFTR cells (indicative of proper maturation and trafficking through the ER). These data were consistent with western blot data that suggested that CFTR expressed in wild-type HSV-CFTR was fully glycosylated while endogenous mutant CFTR was non-glycosylated (FIG. 3).

[00154] Finally, the functionality of human CFTR expressed by HSV-CFTR in SAECs from CF patients infected was confirmed using a dihydrorhodamine 6G (dR6G) fluorescent dye uptake assay that was previously validated as a functional endpoint for virus-mediated CFTR restoration in CF 2D patient epithelial cell culture (Wersto, 1996, Proc Natl Acad Sci USA, 93(3), pp. 1167-72). In summary, HSV-CFTR or SAECs from CF patients Petition 870260061885, dated 06 / 24 / 2026, pp. 103 / 151 / 101 primary with simulated infection were incubated with cell culture medium containing dR6G for 15 minutes, washed four times with PBS, lysed in RIPA buffer, and the 526 nm excitation / 555 nm emission fluorescence was read for each sample using a plate reader. dR6G is itself non-fluorescent, but is converted into the fluorescent compound rhodamine 6G upon cellular uptake and exposure to intracellular dehydrogenases, a process that depends on the presence of functional CFTR (Wersto, 1996, Proc Natl Acad Sci USA, 93(3), pp. 1167-72). A BCA assay was performed on each cell lysate to quantify the total protein content, and the relative fluorescence per pg of total protein was calculated for each sample (FIG. 5).Infection with HSV-CFTR of primary SAECs from CF patients caused a modest, dose-dependent increase in dR6G uptake compared to sham infection controls, indicating that HSV-CFTR was able to restore CFTR function in these diseased primary epithelial cells. Example 3: In vitro dose variation of HSV-CFTR and pharmacology in 3D organotypic cultures using organoids derived from CF patients

[00155] Mutations in the CFTR gene are classified into one of six classes by the primary mechanism that leads to CFTR malfunction. Mutations affecting synthesis and processing result in more severe disease because little or no protein reaches the cell surface; mutations that do not interfere with luminal trafficking but reduce CFTR-mediated anion efflux often lead to less severe symptoms due to the retention of some residual CFTR function in the apical membrane (Foundation, 2019, 2018 Annual Data Report, Bethesda: Cystic Fibrosis Foundation). Because CFTR mutations affect distinct stages of protein synthesis and function, recent drug development efforts have focused on small molecule modulating therapies targeting a specific source of the protein defect.For example, ivacaftor,. Petition 870260061885, dated 06 / 24 / 2026, pp. 104 / 151 / 101, subclassified as a CFTR protein enhancer, increases membrane CFTR chloride secretion (providing clinical benefit for people with specific CFTR class III and IV conductance and gating mutations), while elexacaftor, subclassified as a CFTR protein corrector, acts by facilitating proper folding and cellular processing of CFTR that would otherwise be degraded by the endoplasmic reticulum quality control pathway (providing clinical benefit for people with specific CFTR class II trafficking mutations) (Clancy, 2019, Am J Respir Crit Care Med, 186(7), pp. 593-7). Although the recent FDA approval of four of these modulating therapies has been a boon for CF patients who harbor the specific mutations that respond to these drugs, these modulators treat only a subset of the CF population.In particular, the need for effective drug intervention is for patients carrying class I mutations (responsible for ~10% of CF cases worldwide), encompassing structure, splicing, and nonsense mutations that result in severely reduced or absent CFTR expression, as these patients suffer from the most severe and deadly forms of CF (Wilschanski, 2012, Front Pharmacol, 20(3), pp. 1-3).

[00156] Due to the lack of suitable animal models of CF, efficacy studies in air-liquid interface differentiated bronchial epithelial cells derived from lung explant materials from CF patients have been used for some drug development efforts after proof-of-concept experimentation in heterologous 2D cell systems (Neuberger, 2011, Methods Mol Biol, 741(1), pp. 39-54) (Randell, 2011, Methods Mol Biol, 742(1), pp. 285-310). However, the limited availability of lung explant tissues and the invasive procedures required to obtain bronchial cells from end-stage disease-free CF patients have led to the development of 3D organotypic systems derived from readily available tissues harvested from CFTR mutant patients. Petition 870260061885, dated 06 / 24 / 2026, pp. 105 / 151 / 101 to test new therapies to treat CF. One such technology, using a forskolin-induced swelling (FIS) assay, employs intestinal organoids derived from CF patients (PDOs) to study the function of the CFTR protein alone or in response to pharmaceutical intervention (Dekkers, 2013, Nat Med, 19(7), pp. 939-45), and has proven to be a breakthrough in drug development for CF. When exposed to forskolin, the organoids rapidly increase their cyclic AMP content, which in turn results in the opening of the CFTR channel.Organoids derived from biopsies taken from healthy individuals swell as a consequence of the transport of ions and water into the organoid lumen mediated by CFTR, while organoids derived from biopsies of CFTR mutant patients (or wild-type organoids exposed to specific pharmacological inhibition of CFTR protein function) have reduced or completely inhibited swelling capacity (Boj, 2017, J Vis Exp, 120(1), p. e55159). The use of CF PDOs allows for the quantitative measurement of CFTR protein function (through the detection of organoid swelling) after treatment with new therapies, and the positive results of this 3D organotype system have been shown to correlate directly with clinical benefit, including both changes in pulmonary responses and sweat chloride concentration in CF patients treated (Berkers, 2019, Cell Rep, 26(7), pp. 1701-1708).

[00157] The following example describes experiments showing that the recombinant HSV-1 HSV-CFTR vector, characterized in Example 2 above, was able to rescue the cystic phenotype of CF PDOs, independently of the underlying CFTR mutation.

[00158] The ability of HSV-CFTR to restore functional CFTR expression was tested in clinically relevant 3D organotype cultures using intestinal organoids derived from four patients with different CF; (1) a female patient homozygous for a Petition 870260061885, dated 06 / 24 / 2026, p. 106 / 151 / 101 F508del CFTR mutation (class II mutation), (2) a male patient also homozygous for the F508del mutation, (3) a female patient homozygous for a G542X nonsense CFTR mutation (class I mutation), and (4) a female patient homozygous for a W1282X nonsense CFTR mutation (class I mutation). To assess CFTR activity in transduced organoids, organoid morphology and size were evaluated 24 or 48 hours post-infection, and an FIS assay was performed as previously described (Boj, 2017, J Vis Exp, 120(1), p. e55159). For efficient infection of CF organoids, the organoids were cut into small fragments, incubated in HSVCFTR solution at the indicated MOIs for 1 hour, and seeded in 96-well clear-bottom plates for analysis. The FIS assay was conducted 24 or 48 hours after seeding, as described in more detail below.

[00159] First, G542X / G542X PDO was infected in MOIs of 10, 20, and 40 to assess the vector's impact on organoid swelling and cell viability. Intestinal organoids derived from a healthy patient were seeded in parallel as a comparator. Surprisingly, organoids transduced with HSV-CFTR showed lumen formation and a clear cystic morphology mimicking wild-type PDOs 24 hours post-infection, suggesting complete functional correction of the diseased phenotype by the engineered vector prior to forskolin addition (FIG. 6A). An HSV vector expressing mCherry was used as a negative control to show that the changes in PDO morphology observed in HSV-CFTR-treated samples were not due to a non-specific response to viral infection. Next, a FIS assay was performed 48 hours post-infection.At t = 0, before the addition of forskolin and subsequent activation of CFTR, organoids transduced with HSV-CFTR already possessed a significantly enlarged lumen area compared to organoids treated with vehicle or infected with mCherry, according to [reference needed]. Petition 870260061885, dated 06 / 24 / 2026, pp. 107 / 151 / 101, the 24-hour post-infection observations (FIG. 6B). Interestingly, only a moderate increase in organoid swelling was observed 60 minutes after the addition of forskolin (t = 60) in organoids transduced with HSVCFTR, probably because these organoids were already near their maximum swelling potential before forskolin exposure (FIG. 6C). The G542X / G542X mutation can be (at least partially) corrected by exposure to the aminoglycoside geneticin (G418), which allows translational reading of the nonsense mutation, and G418 was included in this assay as a positive control. While the G542X / G542X PDOs swelled in the presence of G418 at t = 60, the mean organoid size in these positive control samples was significantly smaller than that of the PDOs exposed to HSV-CFTR (FIGS. 6B and 6C).Mild to moderate vector toxicity in G542X / G542X PDOs was observed 48 hours post-infection when HSV-CFTR was used at an MOI of 20 or 40, and the toxicity at an MOI >20 likely accounts for the decreased swelling capacity observed in these organoids compared to samples infected at an MOI of 10. However, although a cytotoxic effect at high MOIs was observed, the treated organoids still outperformed the positive small molecule control.

[00160] Because HSV-CFTR corrected diseased organoids to wild-type morphology (large cystic lumen) in all MOIs tested within 24 hours, and higher doses of HSV-CFTR appeared to negatively impact organoids in swelling assays, the three remaining cystic fibrosis PDOs were tested at lower doses of HSV-CFTR (MOIs 1, 5, and 10) and analyzed using the FIS assay 24 hours post-infection. First, HSV-CFTR was tested in PDOs derived from a patient who is homozygous for the F508del mutation of CFTR. F508del is the most common mutation in cystic fibrosis patients; at least one copy of this allele is found in approximately 85%. Petition 870260061885, dated 06 / 24 / 2026, pp. 108 / 151 / 101 of CF patients worldwide, and F508del is responsible for approximately 70% of CFTR loss-of-function mutations (Maiuri, 2015, Ann Transl Med, 3(Supple 1), p. S24). Most F508del organoid cultures tested showed a cystic morphology (wild type) 24 hours after infection with HSV-CFTR, even at the lowest dose tested (MOI of 1). The average size of F508del organoids treated with HSV-CFTR was significantly increased compared to vehicle control or mCherry-infected organoids before forskolin addition (FIG. 7A). No significant change in average organoid size was detected after the addition of forskolin to samples transduced with HSV-CFTR, since these organoids are already at or near their maximum swelling capacity, i.e., pre-swelled (FIG. 7B).It is important to highlight that the functional correction of the CFTR defect in F508del organoids was considered similar between organoids treated with HSV-CFTR before forskolin treatment and organoids exposed to positive control Orkambi® 60 minutes after forskolin treatment (FIG. 7A vs. FIG. 7B). Orkambi® is an FDA-approved lumacaftor / ivacaftor combination therapy for the treatment of CF patients 2 years of age or older who are homozygous for the F508del mutation. No apparent cytotoxicity attributable to the vector was observed in any of the MOIs tested.

[00161] Subsequently, organoids derived from a patient homozygous for a second nonsense CFTR mutation (W1282X) were infected with HSV-CFTR, and the organoid size was quantified before and after the addition of forskolin. According to the data presented in FIG.6 above, HSV-CFTR efficiently restored the wild-type cystic phenotype and increased the size of the mean organoid 24 hours after infection in nonsense CFTR PDOs W1282X / W1282X before the addition of forskolin (FIG. 8A). Again, HSV-CFTR at an MOI as low as 1 appeared to correct the disease morphology before and after the addition of forskolin (FIGS. Petition 870260061885, dated 06 / 24 / 2026, pp. 109 / 151 / 101 8A and 8B). G418 was also included in these experiments; however, W1282X / W1282X PDOs do not respond to this reading aminoglycoside, therefore no positive control could be included in this experiment (as no effective therapy currently exists for all CFTR nonsense mutations). These data suggested that HSV-CFTR could restore CFTR function in samples from both responsive and non-responsive G418 CFTR null patients.

[00162] Finally, organoids from a second homozygous F508del patient were tested. The HSV-CFTR-infected PDOs had a slightly increased average size compared to the vehicle-treated organoids, but this difference was not statistically significant (FIGS 9A and 9B).

[00163] Data from these studies revealed that transduction of intestinal organoids with CF using HSV-CFTR resulted in a notable alteration of organoid morphology, from a compact budding CF phenotype to a cystic organoid phenotype containing a well-defined lumen exhibiting wild-type characteristics, within 24 hours of infection in MOIs ranging from 1 to 40. This pre-swelled wild-type phenotype was quantitatively demonstrated by measuring the total organoid size, before the addition of forskolin and resulting CFTR activation, compared to multiple negative controls. Due to the pre-swelled nature of the HSV-CFTR-transduced organoids, the ability of forskolin to stimulate further swelling was limited.The observation of a corrected cystic morphology in CF organoids exposed to low doses of HSV-CFTR suggested that high levels of exogenous wild-type CFTR expressed in a minority of cells were sufficient to establish disease correction, indicating a dominant effect of this therapeutic modality. An F508del organoid showed a slightly less efficient restoration of the wild-type phenotype in comparison. Petition 870260061885, dated 06 / 24 / 2026, pp. 110 / 151 / 101, compared with the other CF organoid cultures examined; however, a cystic morphology was observed in all CF organoids infected with HSV-CFTR at an MOI of 5 or higher. The differences observed between the various CF intestinal organoid cultures were likely due to slight alterations in their proliferative or differentiation status at the time of infection, and therefore it is unlikely that the CFTR genotype itself contributed significantly to the efficiency of HSVCFTR transduction or functional CFTR expression. On the other hand, HSV-CFTR corrected the CF disease phenotype independently of the underlying CFTR mutation in this clinically translatable 3D organotype system.

[00164] Taken together, the data provided in these examples indicate that HSV-CFTR infected relevant airway epithelia, effectively produced functional human CFTR, and molecularly corrected several CFTR defects without significant toxicity. Without wishing to be limited by theory, these studies are believed to represent the first example of experimental validation of an attenuated HSV-based gene therapy vector for the delivery of full-length functional human CFTR, supporting the application of HSV-CFTR as a novel gene therapy widely applicable for the treatment of CF. Example 4: In vivo proof-of-concept administration of an inhaled HSV-based vector.

[00165] The following example describes an in vivo proof-of-concept study examining the feasibility of administering an HSV-based vector to the trachea and / or lungs of immunocompetent animals after intranasal or intratracheal administration of the virus.

[00166] All procedures conducted in this example were in accordance with applicable animal welfare laws and were approved by the local Institutional Animal Care and Use Committee (IACUC). 10 five- to six-week-old C57BL / 6 mice were Petition 870260061885, dated 06 / 24 / 2026, pp. 111 / 151 / 101 used in the study, five of which received HSV-mCherry (described above) or vehicle control by intratracheal administration, and five of which received HSV-mCherry or vehicle control by intranasal administration. Prior to the experimental procedures, the animals were sealed with an intraperitoneal injection of a telazol / dexdomitor mixture and an ophthalmic ointment was applied to the eyes to prevent corneal dryness.

[00167] For intratracheal administration, the neck of each mouse was shaved with an electric razor and depilatory cream was applied to remove all remaining hair. The surgical area was then cleaned twice with swabs soaked in 70% ethanol, and the anesthetized mice were positioned in an angled restraint stand. A small incision in the neck was made with surgical scissors, and the thymus, platysma, and anterior tracheal muscles were retracted for visualization and access to the tracheal rings. An intratracheal injection of 25 g / L of 4.9375 x 10⁸ plaque-forming units (PFUs) of HSV-mCherry was administered to three animals, while an intratracheal injection of 25 g / L of vehicle control was administered to two animals, and each mouse was kept in a suspended position until respiration gradually returned to normal. The incision site was closed with simple, individually knotted sutures.

[00168] For intranasal administration, mice were anesthetized as described above and positioned in an angled restraint holder. Three mice were inoculated intranasally with 4.9375x10⁸ PFUs of virus formulated in 25 gL (12.5 gL per nostril). The release rate of the formulation was adjusted to allow the mouse to inhale the inoculum without forming bubbles during the inspiratory phase of respiration. Two mice were administered 25 gL of control vehicle using the same procedure. After administration, the animals were kept in a suspended position until respiration returned to normal. Petition 870260061885, dated 06 / 24 / 2026, pp. 112 / 151 / 101

[00169] All animals were able to recover from anesthesia and received food and water ad libitum until the time of sacrifice. 48 hours after administration, the mice were sacrificed and bronchoalveolar lavage (BAL) was performed on the left and right lungs using sterile saline solution. The BAL fluid was collected, centrifuged, and cell pellets were collected. Then, the upper portions of the trachea were harvested and flash-frozen in liquid nitrogen for nucleic acid quantification. The lungs (left lobe, right upper lobe, right middle lobe, and right lower lobe and post-cavum) were harvested individually and frozen in liquid nitrogen for nucleic acid analysis or perfused in 4% neutral buffered formalin and embedded in paraffin for immunofluorescence analysis.

[00170] For immunofluorescence staining of paraffin-embedded lung tissue, a pan-cytokeratin antibody conjugated with Alexa Fluor® 488 was used to detect epithelial cells (Invitrogen cat. No. 53-9003-82) and a rabbit anti-mCherry primary antibody (Abcam cat. no. ab213511) and a secondary antibody conjugated with Alexa Fluor® 594 (Abcam cat. no. ab150080) to detect infected cells. Tissue samples were mounted on mounting medium containing DAPI to visualize the nuclei.

[00171] Intranasal vs. intratracheal administration of HSV-mCherry resulted in similar levels of mCherry transcripts being detected in the lung tissue of transduced animals (FIG. 10A). Interestingly, while few to no transgene transcripts were identified in the tracheas of intranasally exposed mice, robust mCherry transcription was detected in the tracheas of intratracheally exposed mice, with no statistically significant difference in transgene expression observed between the lungs and tracheas of these invasively treated animals. Furthermore, a higher mean count of Petition 870260061885, dated 06 / 24 / 2026, pp. 113 / 151 100 / 101 total cells per mL of BAL fluid were observed in animals administered intratracheally (646,667 cells / mL and 393,333 cells / mL for intratracheal and intranasal administration, respectively), suggesting a greater influx of inflammatory cells into the lungs after intratracheal administration of the HSV-based vector. Transgene protein expression in lung epithelial tissue was observed in intranasally (FIG. 10B) and intratracheally (FIG. 10C) exposed animals dosed with HSV-mCherry, but not in the corresponding vehicle controls.

[00172] Taken together, these data indicate that an engineered HSV vector can be delivered to the lungs of immunocompetent animals via multiple routes of administration and, moreover, that a non-invasive inhaled route of administration allows for similar levels of transgene expression in the lungs as a more direct and invasive route of administration, while inducing less cellular (inflammatory) invasion. Example 5: HSV-CFTR nebulization

[00173] The following example describes a study examining a nebulizer-based noninvasive delivery route for HSV-CFTR in the airways of immunocompetent wild-type and CFTR-deficient mice.

[00174] Sixteen mice are used in the study: 12 immunocompetent C57BL / 6 animals and 4 immunocompetent gut-corrected CFTR-deficient animals. Table 1 provides a summary of the study. Four wild-type animals are administered HSV-CFTR via intranasal instillation, while the remaining animals are administered HSV-CFTR (or vehicle control) via nebulization (e.g., employing a vibrating mesh nebulizer). Forty-eight hours after dosing, the animals are sacrificed, BAL fluid is collected, and tissue samples along the respiratory tract and lungs are collected, i.e., the upper trachea and Petition 870260061885, dated 06 / 24 / 2026, pp. 114 / 151 101 / 101 inferior, the left and right bronchi, the left lung, and the right lung (superior, middle, inferior, and post-caval lobes, individually). Tissues from two animals / group are frozen in liquid nitrogen and processed for nucleic acid analysis. Vector genomes / 50 ng total DNA are quantified in each tissue by qPCR analysis; human CFTR transcripts / 50 ng total RNA are quantified in each tissue by qRT-PCR analysis. Tissues from the two remaining animals / group are perfused and paraffin-embedded for immunofluorescence / immunohistochemistry. BAL fluid is processed to examine immune cell infiltration in the lungs. Table 1 - Study Design Group Treatment Route n Animals Necropsy 1 Vehicle Inhalation 4 C57BL / 6 48 hours 2 HSV-CFTR Intranasal instillation 4 C57BL / 6 3 HSV-CFTR Inhalation 4 C57BL / 6 4 HSV-CFTR Inhalation 4 CFTRtm1Unc Tg(FABPCFTR) Petition 870260061885, dated 06 / 24 / 2026, pp. 115 / 151

Claims

1. Herpes virus comprising a recombinant herpes virus genome, characterized in that the recombinant herpes virus genome comprises one or more polynucleotides encoding a cystic fibrosis transmembrane conductance regulator (CFTR) polypeptide, wherein the herpes virus is formulated into a nebulizer.

2. Herpes virus, according to claim 1, characterized in that the genome of the recombinant herpes virus comprises one or more polynucleotides encoding the CFTR polypeptide at one or more viral gene loci.

3. Herpes virus, according to claim 1 or claim 2, characterized in that the CFTR polypeptide comprises a sequence with at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% sequence identity with the amino acid sequence of SEQ ID NO: 5 or SEQ ID NO:

6.

4. Herpes virus, according to any one of claims 1 to 3, characterized in that the genome of the recombinant herpes virus is a genome of the recombinant herpes simplex virus.

5. Herpes virus, according to claim 4, characterized in that the genome of the recombinant herpes simplex virus is a recombinant HSV-1 genome.

6. Herpes virus, according to claim 4 or claim 5, characterized in that the genome of the recombinant herpes simplex virus comprises an inactivating mutation in a gene of the herpes simplex virus selected from the group consisting of Infected Cell Protein (ICP) 0, ICP4, ICP22, ICP27, ICP47, thymidine kinase (tk), Petition 870260061885, dated 06 / 24 / 2026, page 116 / 151 2 / 6 Long Single Region (UL) 41 and UL55.

7. Herpes virus, according to claim 6, characterized in that the genome of the recombinant herpes simplex virus comprises an inactivating mutation in one or both copies of the ICP4 gene.

8. Herpes virus, according to any one of claims 1 to 7, characterized in that one or more polynucleotides encoding a CFTR polypeptide are operatively linked to a promoter and expressed from the genome of the recombinant herpes virus.

9. Herpes virus, according to any one of claims 1 to 8, characterized in that the herpes virus is defective in replication.

10. Herpes virus, according to any one of claims 1 to 9, characterized in that the herpes virus is a herpes simplex virus.

11. Herpes virus, according to any one of claims 1 to 10, characterized in that the herpes virus is attenuated.

12. Herpes virus, according to any one of claims 1 to 11, characterized in that the herpes virus is aerosolized by means of nebulization.

13. Herpes virus, according to claim 12, characterized in that the aerosolized herpes virus comprises droplets with diameters less than about 5 microns in mass-mean-aerodynamic diameter.

14. Herpes virus, according to any one of claims 1 to 13, characterized in that the nebulizer is a jet nebulizer, an ultrasonic nebulizer or a vibratory mesh nebulizer. Petition 870260061885, dated 06 / 24 / 2026, page 117 / 151 3 / 6 15. Use of the herpes virus as defined in any of claims 1 to 14, characterized by being for the manufacture of a medicament to enhance, increase, raise and / or supplement the levels of a cystic fibrosis transmembrane conductance regulator (CFTR) polypeptide in one or more cells of a subject in need thereof.

16. A method for intensifying, increasing, raising and / or supplementing the levels of a CFTR polypeptide in one or more cells of a subject, the method characterized in that it comprises administering to the subject an effective quantity of the herpes virus as defined in any one of claims 1 to 14.

17. Use according to claim 15, or method according to claim 16, characterized in that one or more cells are one or more cells of the respiratory tract.

18. Use or method, according to claim 17, characterized in that one or more cells are either one or more epithelial cells of the airways or one or more cells of the submucosal glands.

19. Use of the herpes virus as defined in any one of claims 1 to 14, characterized in that it is for the manufacture of a medicament to reduce or inhibit the progressive destruction of the lung in a subject in need thereof.

20. A method for reducing or inhibiting the progressive destruction of the lung in a subject in need thereof, the method characterized in that it comprises administering to the subject an effective quantity of the herpes virus as defined in any one of claims 1 to 14.

21. Use or method, according to any of claims 15 to 20, characterized in that the subject suffers from a chronic lung disease. Petition 870260061885, dated 06 / 24 / 2026, p. 118 / 151 4 / 6 22. Use or method, according to claim 21, characterized in that the chronic lung disease is cystic fibrosis or chronic obstructive pulmonary disease (COPD).

23. Use of the herpes virus as defined in any of claims 1 to 14, characterized in that it is for the manufacture of a medicament to provide prophylactic, palliative or therapeutic relief of one or more signs or symptoms of cystic fibrosis in a subject in need thereof.

24. A method for providing prophylactic, palliative or therapeutic relief of one or more signs or symptoms of cystic fibrosis in a subject in need thereof, the method characterized in that it comprises administering to the subject an effective quantity of the herpes virus as defined in any one of claims 1 to 14.

25. Use according to claim 23 or method according to claim 24, characterized in that one or more signs or symptoms of cystic fibrosis are selected from the group consisting of a persistent cough producing thick mucus, thick and sticky mucus that accumulates in the airways, wheezing, shortness of breath, sinusitis, repeated lung infections, inflamed nasal passages, bronchiectasis, nasal polyps, hemoptysis, pneumothorax, pancreatitis, recurrent pneumonia, respiratory failure and any combinations thereof.

26. Use of the herpes virus as defined in any of claims 1 to 14, characterized in that it is for the manufacture of a medicament to provide prophylactic, palliative or therapeutic relief of one or more signs or symptoms of COPD in a subject in need thereof.

27. Method for providing prophylactic, palliative or therapeutic relief of one or more signs or symptoms of COPD in a subject in need thereof, the method characterized in that it comprises administering to the subject an effective amount of the herpes virus as defined in any one of claims 1 to 14.

28. Use according to claim 26 or method according to claim 27, characterized in that one or more signs or symptoms of COPD are selected from the group consisting of shortness of breath, wheezing, chest tightness, excess mucus in the lungs, chronic cough, cyanosis, frequent respiratory infections and any combinations thereof.

29. Use or method, according to any one of claims 15 to 28, characterized in that the subject is a human.

30. Use or method, according to any one of claims 15 to 29, characterized in that the subject's genome comprises a loss-of-function mutation in a CFTR gene.

31. Use or method, according to any one of claims 15 to 30, characterized in that the herpes virus is administered to the subject orally, intranasally, intratracheally or by inhalation.

32. Use or method, according to any one of claims 15 to 31, characterized in that the herpes virus is administered to the subject by inhalation.

33. Use or method, according to any one of claims 15 to 32, characterized in that the herpes virus is administered by means of non-invasive inhalation administration.

34. Use or method, according to any one of claims 15 to 33, characterized in that the herpes virus is administered using a dry powder inhaler, a pressurized metered-dose inhaler, a soft mist inhaler, a nebulizer, or an electro-hydrodynamic aerosol device.

35. Use or method, according to any of the claims 15 to 34, characterized in that the herpes virus is administered using a nebulizer.

36. Use or method according to claim 35, characterized in that the nebulizer is a jet nebulizer, an ultrasonic nebulizer or a vibrating mesh nebulizer.

Images