DETAILED ACTION
Notice of Pre-AIA or AIA Status
1. The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Priority
2. The instant application, filed on 03 May, 2023, claims domestic benefit to US provisional application no. 63/340,447, filed on 10 May, 2022.
Status of Application, Amendments, and/or Claims
3. The response filed on 3 May 2023 has been entered in full. No amendments or withdrawals have been made. Therefore, claims 1- 20 are pending and are the subject of this Office Action.
Specification
4. The disclosure is objected to because of the following informalities:
In paragraph 00177 on page 21 line 22 reads “NL cells”
In paragraph 00178 on page 22 line 7 reads “in one embodiment NK cells are expressing an enhanced concentration of NK cells.”
Appropriate correction is required.
Claim Objections
5. Claims 2-4, and 8 are objected to because of the following informality: the loss of plurality in the term natural killer cell. Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
6. Claims 6, 12, and 16 are rejected under 35 U.S.C. 112(b), as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor, regards as the invention.
7. Claim 6 recites the limitation “said enhanced survival and/or activity" in line 1. There is insufficient antecedent basis for this limitation in the claim. In light of the specification which states “Said “activation” of NK cells by hypoxia is associated with induction of cell protective enzymes such as heme-oxygenase, superoxide dismutase and NFR-2” and “NK cells can be exposed to low oxygen conditions under any methodology that permits the NK cells to attain an enhanced differentiation potential, proliferation rate, engraftment ability and/or in vivo tumor migratory ability”, enhanced survival and/or activity will be interpreted as induction of cell protective enzymes for enhanced differentiation potential, proliferation rate, engraftment ability and/or in vivo tumor migratory ability.
8. Claim 12 recites the limitation "T-Cells are generated to possess a hypoxia resistant phenotype instead of NK cells" in line 1. There is insufficient antecedent basis for this limitation in the claim due to the independent claim for which claim 12 relies only speaking to NK cells in the method and thus T-Cells makes the claim indefinite. The claim is interpreted as “NK cells are generated to possess a hypoxia resistant phenotype.”
9. Claim 16 is rejected under 35 U.S.C. 112(b), as being unclear. Claim 13 from which claim 16 depends states “NK cells are cultured with regenerative cells.” Claim 16 states “addition of culture supernatant from said regenerative cells to said NK cells”. It is unclear if the conditioned media of the regenerative cells is added to a NK cell monoculture or to a co-culture of the regenerative cells and NK cells which is essential to the definiteness of the claim. In light of the specification which states “a therapeutic composition comprising a population of human hematopoietic stem or progenitor cells which have been differentiated into NK cells possessing regenerative properties by cultured in conditioned media of mesenchymal stem cells, wherein said cells are suspended in a sterile, therapeutically acceptable solution suitable for administration to a patient.” (paragraph 00177, page 21, line 22-24) the claim will be interpreted as culturing NK cells monoculture in regenerative cell conditioned media.
The following is a quotation of 35 U.S.C. 112(d):
(d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
10. Claim 2 is rejected under 35 U.S.C. 112(d), as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends. The statement “natural killer cells are either autologous, allogenic, or xenogeneic to said patient” does not further limit the scope of claim 1 because the three options listed are the only three classes of cellular therapeutic source types. Applicant may cancel the claim, amend the claim to place the claim in proper dependent form, rewrite the claim in independent form, or present a sufficient showing that the dependent claim complies with the statutory requirements.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
11. Claims 1-3 and 13-16 are rejected under 35 U.S.C. 103 as being unpatentable over Thomas et al. (2014) Interaction with Mesenchymal Stem Cells Provokes Natural Killer Cells for Enhanced IL-12/IL-18-Induced Interferon-Gamma Secretion Mediators of Inflammation 143463 (hereafter Thomas) in view of Nelson (2001) Autoimmune Ovarian Failure: Comparing the Mouse Model and Human Disease the Journal of the Society for Gynecological Investigation 8: S55-S57 (hereafter Nelson).
12. In regards to claim 1, Thomas teaches natural killer (NK) cells which have been modified to allow for enhanced regenerative activity through enhanced IFN-γ production wherein said modification comprises contacting said NK cells with Mesenchymal Stem Cells (MSCs) (a regenerative cell line) (section 2.3, paragraph 1). Thomas also further suggests “NK cells play an important role in the repair and regeneration of damaged tissue” (section 1, paragraph 3) and “IFN- γ also plays a functional role in the process of tissue regeneration” (section 1, paragraph 2).
Thomas fails to teach a method of treating ovarian degeneration, however, Nelson teaches that in women with premature ovarian failure (POF) (a type of ovarian degeneration initiated by chemotherapy) exhibit decreased NK cell activity disrupting the Th1/Th2 balance needed for ovarian function regulation.
Thus, Thomas teaches a method of modifying NK cells through contact with MSCs to achieve enhanced regenerative activity, and Nelson teaches that women with POF exhibit decreased NK cell activity. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to try the teachings of Thomas in light of the teachings of Nelson with a reasonable expectation of success to improve the NK population and activity in a patient as a method to treat ovarian degeneration.
13. In regards to claims 2 and 3 Thomas further teaches the use of primary human NK cells or NK-92 (section 2.2, line 1) cells for the use of contact with regenerative cells which could be autologous, allogeneic, or xenogeneic depending on the subject receiving the treatment.
14. In regards to claims 13-16, Thomas teaches NK cells cultured with Mesenchymal stem cells both through direct contact and transwell experiments (section 2.3, paragraph 1, line 4). Thomas also teaches co-culture of the NK cells and regenerative cells in the presence of IL-12 (section 2.3, paragraph 1 lines 7-9) or cell culture supernatant from independently cultured regenerative cells (section 2.3, paragraph 3, lines 13-14). Thomas further teaches that IL-15 as an option to potentiate IL-12 activity and subsequently IFN-γ production (section 1, paragraph 2, lines 13-14).
Thus, Thomas discloses a method for creating NK-92 cells with enhanced regenerative activity through contact with mesenchymal stem cells population by co-culture to induce the production of IFN-γ both in the presence or in the absence of IL-12 or IL-15. Nelson teaches women with POF exhibit decreased NK cell activity disrupting the Th1/Th2 balance needed for ovarian function regulation. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to try the teachings of Thomas in light of the teachings of Nelson with a reasonable expectation of success to get a method to treat ovarian degeneration using NK cells modified by contact with a regenerative cell population as a cell transplantation therapy.
15. Claims 4 and 5 are rejected under 35 U.S.C. 103 as being unpatentable over Thomas in view of Nelson as applied to claim 1 above, and further in view of Hermanson et al. (2016) Induced pluripotent stem cell-derived natural killer cells for treatment of ovarian cancer Stem Cells 34(1) 93-101 (hereafter Hermanson).
Thomas and Nelson fail to teach the use of inducible pluripotent stem cells, however, Hermanson teaches the benefit of using iPSC derived NK cells because they “can be utilized as an allogenic ‘off the shelf’ immunotherapy” with the feasibility of repeated doses due to high yields of product (Introduction, paragraph 3, lines 7-9).
Thus, Hermanson teaches the benefits of iPSC derived NK cells for higher product yield and Thomas in view of Nelson teach the methods of using said NK cells as outlined above. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to combine the teachings of Thomas in view of Nelson, and Hermanson with a reasonable expectation of success to get the of a method to treat ovarian degeneration using NK cells wherein the NKs are derived from iPSC and modified by contact with a regenerative cell population as a cell transplantation therapy.
16. Claims 6,7 and 12 are rejected under 35 U.S.C. 103 as being unpatentable over Thomas in view of Nelson as applied to claim 1 above, and further in view of Lim et al. (2021) Hypoxia-Driven HIF-1α Activation Reprograms Pre-Activated NK Cells towards Highly Potent Effector Phenotypes via ERK/STAT3 Pathways Cancers 13(8) 1904 (hereafter Lim) and Hertzog et al. (2020) Hypoxic preconditioning — A nonpharmacological approach in COVID-19 prevention International Journal of Infectious Diseases (103) 415-419 (hereafter Hertzog).
In regards to claims 6 and 7, Thomas in view of Nelson fails to teach the use of hypoxic preconditioning in which HIF-1α expression is increased to enhance cell survival and/or activity.
Lim, however, teaches natural killer cells preactivated with IL-2 when exposed to hypoxic conditions showed increased the expression of HIF-1α by more than 25% (Figure 4B) and exhibited increased proliferation and downregulation of cell cycle arrest and cell death genes (Figure 3E).
In regards to claim 12 Hertzog defines hypoxic preconditioning as “the exposure of organisms, organs, tissues, or cells to non-injurious, repetitive mild or moderate hypoxic episodes, which result in increased tolerance and cell protection against subsequent severe hypoxia exposures and other stresses” (page 416, Col 1, lines 60-61 and Col 2, lines 1-3 ) which is described by the process in claims 6 and 7, therefore claim 12 is rejected under the same references as claims 6 and 7 further supported by Hertzog’s definition of hypoxic preconditioning.
Thus, Thomas in view of Nelson teaches a method of using modified NK cells by contacting said NK cells with a regenerative cell line as a treatment for ovarian degeneration as outlined above and Lim teaches a method where hypoxic conditions increased HIF-1α expression by at least 25% and enhanced the NK cells survival and activity. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to combine the teachings of Thomas and Nelson, and incorporate the teachings of Lim with a reasonable expectation of success to get the method of treating ovarian degeneration using NK cells wherein the NKs have enhanced survival and/or activity from hypoxic preconditioning and increased regenerative activity by contact with a regenerative cell population as a cell transplantation therapy.
17. Claims 8-11 are rejected under 35 U.S.C. 103 as being unpatentable over Thomas in view of Nelson as applied to claim 1 above, and further in view of Ogbomo et al. Histone deacetylase inhibitors suppress natural killer cell cytolytic activity (2007) FEBS Letters 581(7) 1317-1322 (hereafter Ogbomo) and Cui et al. Human mesenchymal stromal/stem cells acquire immunostimulatory capacity upon cross-talk with natural killer cells and might improve the NK cell function of immunocompromised patients (2016) Stem Cell Research & Therapy 7:88 (hereafter Cui).
In regards to claims 8 -11, Thomas in view of Nelson fails to teach the use of an epigenetic modulator to further modify the NK cells. However, Ogbomo teaches that treatment of NK cells with a histone deacetylase inhibitor (an epigenetic modulator) decreases cytotoxic effects of the NK cells (section 4, paragraph 1, lines 6-9). Ogbomo further teaches the major classes of histone deacetylase inhibitors short fatty acids, derivative of hydroxamic acid, Benzamide derivatives, which includes trichostatin A and mocetinostat (section 1, paragraph 1, lines 3-7). Ogbomo fails to teach how this would enhance the survival or activity of the NK cells, however, Cui teaches that NK cells with poor cytotoxicity release high amounts of IFN-γ upon exposure to IL-12 (page 2, col 1, lines 38-40) which is further supported by Thomas’ teachings of the importance of IFN-γ in co-culture with MSCs to drive NK cell regenerative activity.
Thus, Thomas in view of Nelson teaches a co-culture method to improve NK cell regenerative activity by IFN-γ activity and the important of NK activity in POF, and Ogbomo teaches decreasing NK cytotoxic effects using histone deacetylase inhibitors which Cui supports by teaching decreased cytotoxicity increases the NK cell production of IFN-γ. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to modify Thomas in view of Nelson to incorporate the teachings of Ogbomo informed by teachings of Cui with a reasonable expectation of success with the motivation of using an epigenetic modulator such as a histone deacetylase to further improve IFN-γ production in NK which increases activity and survival for the purposes of ovarian regeneration.
18. Claims 17-19 are rejected under 35 U.S.C. 103 as being unpatentable over Thomas in view of Nelson as applied to claims 1 and 13 above, and further in view of Cui.
In regards to claims 17-19 Thomas in view of Nelson fails to teach regenerative cell activation by an inflammatory cytokine. However, Cui teaches that stimulation of a MSCs (a regenerative cell line) with IFN-γ (an inflammatory cytokine) to increase the production of CCL2 (Figure 6). Cui also teaches that NK cells and MSCs have a positive feedback loop in which MSCs prime NK cells to produce IFN-γ which increases CCL2 production in MSCs which further pushes NK cell production of IFN-γ when in co-culture. A feedback loop which is enhanced with the pre activation of MSCs with IFN- γ to get CCL2 rich medium (page 3, column 2, lines 35-37 and page 12, column 1 lines 48-55). Cui further teaches “MSC/NK cell co-cultures from healthy donors improved the IFN- γ production of the patients’ NK cells in a CCR2-dependent manner” (the receptor of CCL2) (Abstract).
Cui fails to teach the NF-κB activation of regenerative cells and a method for treating ovarian degeneration. However, Thomas teaches that IFN- γ transcription requires NF-κB activation, and that the IFN- γ secretion plays a functional role in the process of tissue regeneration (section 1, paragraph 2, lines 20-22 and 31-34).
Thus, Cui teaches the benefits of pre activation of a regenerative cell line with the inflammatory cytokine IFN- γ increases the regenerative cells production of CCL2 to further mediate NK regenerative activity through IFN- γ production which is supported by Thomas’ teachings of the importance of the activation of NF-κB for the stimulation of IFN- γ production and the role of IFN- γ in tissue regeneration. Further Thomas in view of Nelson teaches a co-culture method to improve NK cell regenerative activity by IFN-γ activity and the deficiency of NK activity in patients with POF. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to combine the teachings of Thomas and Cui supported by the teachings of Nelson with a reasonable expectation of success to activate the regenerative cells with an inflammatory cytokine which stimulate the NF-κB pathway to increase IFN-γ production to improve the effects of the regenerative cells on NK cells in co-culture to develop a method to treat ovarian degeneration.
19. Claim 20 is rejected under 35 U.S.C. 103 as being unpatentable over Thomas and Nelson as applied to claims 1 and 13 above, and further in view of Chrysanthopoulou et al. (2014) Neutrophil Extracellular traps promote differentiation and function of fibroblast Journal of Pathology 233: 294-307 (hereafter Chrysanthopoulou) and Huang et al. (2009) IL-17 stimulates the proliferation and differentiation of human mesenchymal stem cells: implication for bone remodeling Cell Death & Differentiation 16, 1332-1343 (hereafter Huang).
Thomas in view of Nelson fails to teach activation of the regenerative cells by neutrophil extracellular traps (NETs), however, Chrysanthopoulou teaches that neutrophil extracellular traps (NETs) contain IL-17 which allows them to increase the proliferation and migration of fibroblast; a mesenchymal, regenerative cell line (abstract).
Chrysanthopoulou fails to teach how this would improve the regenerative activity of natural killer cells, however Huang teaches that IL-17 induces receptor activator of NF-κB ligand (RANKL) expression in human mesenchymal stem cells (abstract) as well as stimulates proliferation and differentiation (abstract). Which Thomas further supports by teaching that IFN- γ transcription requires NF-κB activation as explained above.
Thus, Chrysanthopoulou teaches the influence of IL-17 expressed in NETs plays a role in mesenchymal cell activation which is further supported by Huang which teaches that IL-17 induces receptor activator of NF-κB ligand (RANKL) in human mesenchymal stem cells as well as stimulates proliferation and differentiation. Thomas teaches the importance of the activation of NF-κB for the stimulation of IFN- γ production and the role of IFN- γ in tissue regeneration. Further Thomas in view of Nelson teaches a co-culture method to improve NK cell regenerative activity by IFN-γ activity and the deficiency of NK activity in patients with POF. Therefore, a person of ordinary skill in the art before the effective filing date of the claimed invention would have found it obvious to incorporate the teachings of Chrysanthopoulou with Thomas in view of Nelson and informed by the teachings of Huang, with a reasonable expectation of success activate the regenerative cells with NETs to stimulate the NF-κB pathway to improve the effects of the regenerative cells on NK cells in co-culture to develop a method to treat ovarian degeneration.
Conclusion
20. Any inquiry concerning this communication or earlier communications from the examiner should be directed to DASIA A ALDARONDO whose telephone number is (571)272-1977. The examiner can normally be reached on Monday through Thursday 7:00-4:30pm or Fridays 7:00-11am.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Joanne Hama, can be reached at telephone number (571)272-2911. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/D.A.A/Examiner, Art Unit 1647 /JOANNE HAMA/Supervisory Patent Examiner, Art Unit 1647