DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Claim Status
Claims 1-20 and 22 have been cancelled and claim 21 has been amended, as requested in the amendment filed on 12/02/2025. Following the amendment, claims 21 and 23-28 are pending in the instant application.
Claims 21 and 23-28 are under examination in the instant office action.
Priority - Updated
Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged.
The later-filed application must be an application for a patent for an invention which is also disclosed in the prior application (the parent or original nonprovisional application or provisional application). The disclosure of the invention in the parent application and in the later-filed application must be sufficient to comply with the requirements of 35 U.S.C. 112(a) or the first paragraph of pre-AIA 35 U.S.C. 112, except for the best mode requirement. See Transco Products, Inc. v. Performance Contracting, Inc., 38 F.3d 551, 32 USPQ2d 1077 (Fed. Cir. 1994).
Upon further consideration, the disclosure of the prior-filed application, Provisional Application No. 63/141,555, fails to provide adequate support or enablement in the manner provided by 35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112, first paragraph for one or more claims of this application.
The prior filed application fails to provide adequate support for instant claim 21, and subsequently dependent claims 23-28, because the prior filed application fails to disclose the instantly claimed method. While it is acknowledged that components of the claimed method are described, they are described separately and their combination is neither taught nor suggested by the prior filed disclosure. For example, the role of MSCs in inducing Tregs is disclosed, administering MSCs in cases of kidney failure/injury/ischemia are disclosed, administering reprogrammed immune cells contacted with regenerative cells are disclosed, the effects of IFN-y on specific MSCs are disclosed, and the role of IL-2 on the induction of Tregs by certain MSCs are disclosed, but at no point is there disclosed the claimed method comprising extracting Tregs from a subject, culturing the Tregs with IFN-y treated MSC-CM (of placental tissue, amniotic membrane, umbilical cord tissue, fallopian tube tissue, and subepithelial umbilical cord tissue), and administering the cultured Tregs to said patient, wherein said administration inhibits and/or reverses kidney failure associated with ischemia and fibrosis. The only working example and explicitly defined method disclosed is drawn to the administration of bone-marrow MSCs (BM-MSCs) or syngenic splenocytes exposed to allogenic BM-MSCs, which demonstrated renal protection based on measured creatinine levels relative to controls (see Example on Page 75; Figure 1). Thus, the prior filed application fails to provide adequate support for instant claim 21, and subsequently dependent claims 23-28.
As such, claims 21 and 23-28 have an effective filing date of January 26, 2022, which is the date the instant application was filed.
Claim Objections - Withdrawn
Claim 22 was objected for a minor informality. Claim 22 has been cancelled, rendering the objection moot. As such, the objection to claim 22 is withdrawn,
Claim Rejections - 35 USC § 112 - Withdrawn
Claims 21-28 were rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, regarding scope of enablement. Independent claim 21 has been amended such that the claims now recites “[a] method of inhibiting, and/or reversing kidney failure associated with ischemia and renal fibrosis”. As such, the rejection of claims 21-28 under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, regarding scope of enablement is withdrawn.
Claim Rejections - 35 USC § 103 - Withdrawn
Claims 21, 25, and 26-28 were rejected under 35 U.S.C. 103 as being unpatentable over US 2017/0239293 A1 (previously cited on PTO-892; herein after referred to as "Ichim") in view of non-patent literature by Silva-Carvalho et. al. (Cytokine and Growth Factor Reviews, 2019, 47, 32-42; previously cited on PTO-892; herein after referred to as "Silva") and non-patent literature by Gandolfo et. al. (Kidney International, 2009, 76, 717-729; previously cited on PTO-892; herein after referred to as "Gandolfo") as evidenced by non-patent literature by Hirakawa et. al. (Blood, 2015, 126(23), 1-4; previously cited on PTO-892; herein after referred to as "Hirakawa").
Claim 22 was rejected under 35 U.S.C. 103 as being unpatentable over Ichim, Silva, and Gandolfo, as evidenced by Hirakawa, as applied to claims 21, 25, and 27-28 above, and further in view of non-patent literature by Sharma and Kinsey (Am. J. Physiol. Renal Physiol., 2018, 314, F679-F698; previously cited on PTO-892; herein after referred to as "Sharma").
Claims 23-24 were rejected under 35 U.S.C. 103 as being unpatentable over Ichim, Silva, Gandolfo, and Sharma, as evidenced by Hirakawa, as applied to claims 21-22, 25, and 27-28 above, and further in view of non-patent literature by Huang et. al. (Scientific Reports, 2015, 5(16565), 1-8; previously cited on PTO-892; herein after referred to as "Huang") and non-patent literature by Levey and Coresh (Lancet, 2012, 379, 165-180; previously cited on PTO-892; herein after referred to as “Levey”).
It is noted that claim 22 has been cancelled, rendering the above-listed rejection in view of Ichim, Silva, Gandolfo, Hirakawa, and Sharma moot. Upon further consideration, and in view of Applicant’s arguments regarding the above-listed combination of references, the above-listed claim rejections are withdrawn.
Specification - New Objections
The disclosure is objected to because of the following informalities: at various points in the instant disclosure, sentences are incomplete. See, for example, Paragraphs 00552, 00558, 00564, 00576, 00578, among others. Appropriate correction is required.
Additionally, the use of the terms, for example, Xigris, FACS, Ficoll-Plaque, etc., which are trade names or marks used in commerce, have been noted in this application. The term should be accompanied by the generic terminology; furthermore the term should be capitalized wherever it appears or, where appropriate, include a proper symbol indicating use in commerce such as ™, SM , or ® following the term.
Although the use of trade names and marks used in commerce (i.e., trademarks, service marks, certification marks, and collective marks) are permissible in patent applications, the proprietary nature of the marks should be respected and every effort made to prevent their use in any manner which might adversely affect their validity as commercial marks.
Claim Rejections - 35 USC § 112 - New
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.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 23, 24, and 25 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
It is noted that as currently presented, claims 23 and 24 depend from claim 22 which is cancelled. As such, the dependency of claims 23 and 24 are unclear and the claim is considered to be indefinite. For the purposes of examination, claims 23 and 24 are being interpreted as depending from instant claim 21.
With regard to claim 25, the claim recites “wherein the MSC-CM expresses higher levels of hepatocyte growth factor (HGF) than normal”. It is noted the instant specification provides no point of comparison for higher expression “than normal” and there is no definition of “normal” nor is it indicated what the “normal” point of comparison would be (i.e., cell type, conditioned media components, etc.) and as such the claim is considered to be indefinite as one of ordinary skill in the art would not be able to ascertain the point for comparison and therefore could not determine the metes and bound of the claim.
Claim Rejections - 35 USC § 103 - New
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 21 and 27 are rejected under 35 U.S.C. 103 as being unpatentable over non-patent literature by Kim et. al. (J. Am. Soc. Nephrol., 2013, 24, 1529-1536; herein after referred to as "Kim") in view of non-patent literature by Landwehr-Kenzel et. al. (Kidney International, 2018, 93, 1452-1464; herein after referred to as "Kenzel"), non-patent literature by Chang et. al. (Stem Cells, 2006, 24(11), 1466-2477; herein after referred to as "Chang"), non-patent literature by Yi et. al. (Cellular Immunology, 2018, 326, 42-51; herein after referred to as "Yi"), and non-patent literature by Gandolfo et. al. (Kidney International, 2009, 76(7), 717-729; previously cited on PTO-892; herein after referred to as “Gandolfo”).
Kim discloses that regulatory T cells (Tregs) can suppress immunologic damage in renal ischemia reperfusion injury (IRI), but the isolation and ex vivo expansion of these cells for clinical application remains challenging; Kim investigated if the IL-2/anti-IL-2 complex (IL-2C), which mediates Treg expansion, could attenuate IRI in mice (Abstract). IL-2C administered before bilateral renal IRI induced Treg expansion in both spleen and kidney, improved renal function, and attenuated histologic renal injury and apoptosis after IRI; IL-2C administration reduced the expression of inflammatory cytokines and attenuated the infiltration of neutrophils and macrophages in renal tissue; depletion of Tregs with anti-CD25 antibodies abrogated the beneficial effects of IL-2C. However, IL-2C–mediated renal protection was not dependent on either IL-10 or TGF-β wherein, notably, IL-2C administered after IRI also enhanced Treg expansion in spleen and kidney, increased tubular cell proliferation, improved renal function, and reduced renal fibrosis; the results of the instant study indicate that IL-2C induced Treg expansion attenuates acute renal damage and improves renal recovery in vivo, suggesting that IL-2C may be a therapeutic strategy for renal IRI (Id.). Kim indicates IL-2C treatment after IRI induced significant expansion of Foxp3+CD4+ Tregs in both spleen and kidney and that the IL-2C group demonstrated renal tubular cell proliferation was significantly increased on day 5 (P=0.04; Figure4 E), and renal fibrosis also significantly decreased on day 28 after IL-2C treatment in the unilateral model (See Page 1533). Thus, Kim discloses promoting in vivo induction/expansion of Tregs in order to attenuate renal ischemia reperfusion injury, and indicates that isolation and expansion of Tregs ex vivo for direct clinical application needs improvement.
Kim does not teach isolating and expanding autologous Treg cells ex vivo in the presence of MSCs (e.g., placental MSCs), IFN-γ, and IL2 prior to the administration of said Tregs to a patient.
One solution to the issue of ex vivo isolation and expansion of Tregs for direct therapeutic application is disclosed by Kenzel. Kenzel discloses an investigation of regulatory T-cells in dialysis and kidney transplanted patients and the feasibility of generating a clinically useful nTreg product from these patients (Abstract). Results of the study indicate that the generation of autologous nTreg products from patients with end stage renal disease (i.e., renal failure) is feasible; 50 ml of blood was peripherally collected from patients with ESRD (healthy volunteers served as independent controls (Figure5a–e)) wherein nTregs were isolated by CliniMACS and polyclonally expanded under Good Manufacture Practice clean room conditions from patients with ESRD and from HCs and the nTregs from patients with ESRD expanded rapidly and reached cell numbers partially exceeding the yield of nTreg cultures from healthy volunteers (Figure5a), the expansion rates, yield, and purity were comparable between all patients with ESRD independent of the primary renal disease, and the end-product purity, defined as CD25highFoxP3+ cells within the end product was comparable to nTreg cultures from HCs and previously published data (Page 1456, Column 1). The suppressive capacity of the nTreg product was consistent with data from healthy donors reported previously (Figure5e); overall the data demonstrates that the generation of autologous nTregs from dialysis-dependent ESRD patients is feasible and safe (Page 1456, Column 2, First Partial Paragraph). Specifically, the protocol for isolation and expansion comprises: (1) nTregs isolation from PBMCs of freshly collected blood by CliniMACS isolation (Miltenyi Biotec); (2) performing CD8 depletion followed by CD25+ enrichment; (3) expansion wherein nTregs were suspended in X-VIVO15 medium (Lonza, Cologne, Germany) containing10% serum, 500U/ml human recombinant IL-2 (Chiron Behring, Marburg, Germany), and 100 nM rapamycin (Sigma-Aldrich), rested overnight, and stimulated with (CD3/CD28) Treg expansion beads (Miltenyi Biotec) (Page 1460; Methods: nTreg Isolation and Expansion).
Kim nor Kenzel disclose culturing autologous Treg cells ex vivo in the presence of MSCs (e.g., placental MSCs) contacted with IFN-γ.
Chang discloses that several types of nonhematopoietic stem cells, including bone marrow mesenchymal stem cells (BMMSCs) and embryonic stem cells, have been shown to have immunosuppressive properties; the authors show that human placenta-derived multipotent cells (PDMCs), which are isolated from a source without ethical concern and harbor multilineage differentiation potential, have strong immunosuppressive properties (Abstract). PDMCs suppressed both mitogen-induced and allogeneic lymphocyte proliferation in both CD4 and CD8 populations wherein the immunosuppression seen with PDMCs was significantly stronger than that with BMMSCs; both PDMCs and BMMSCs express indoleamine 2,3-dioxygenase, but only PDMCs are positive for intracellular human leukocyte antigen-G (HLA) (Id.). Mechanistically, suppression of lymphocyte reactivity by PDMCs is not due to cell death but to decreased cell proliferation and increased numbers of regulatory T cells; unlike BMMSCs, PDMCs treated
with interferon-γ for 3 days only very minimally upregulated HLA-DR whereas, on the contrary, PD-L1, a cell surface marker that plays an inhibitory role in T-cell activation, was upregulated and TGF-expression was seen (Id.). The immune-suppressive properties of PDMCs, along with their multilineage differentiation potential, ease of accessibility, and abundant cell numbers, may render these cells as good potential sources for future therapeutic applications (Id.). Flow cytometric analysis for CD4+CD25high Treg cells after 3 days of incubation with third-party PDMCs revealed a more-than-three-fold increase in the proportion of CD4+CD25high T cells in MLCs stimulated with PHA after 3 days of coculture with PDMCs (Fig. 3C) and, using another Treg marker Foxp3, also found was a threefold increase in CD4+/Foxp3 lymphocytes when third-party PDMCs were added to MLCs (Page 2470, Column 2, Last Paragraph; supplemental online Fig. 2). Reverse transcription-PCR showed a strong increase in the expression of TGF-β in PDMCs after 3 days of IF-γ treatment (Fig. 6B); using CFSE-labeled CD4 and CD8 T cells, MLCs were performed with third-party PDMCs treated either with or without IF-γ, and with the addition of IL-10- and TGF-β-neutralizing antibodies wherein it was observed that IF-γ-treated PDMCs are able to resuppress the lymphocyte proliferation restored by the neutralizing antibodies, whereas untreated PDMCs were not able to do so (Fig. 6C) (Page 2473).
Yi discloses that interferon gamma (IFNγ) suppressed the proliferation and migration for human placenta-derived mesenchymal stromal cells (hPMSCs), and enhanced the ability of said hPMSCs to induce the generation of the CD4+CXCR5+Foxp3+ Treg subset and the maximal effectiveness for IFNγ treated hPMSCs upon inducing the generation of Treg subsets was for CD4+CXCR5+Foxp3+Treg subset as compared with that of CD4+CD25+Foxp3+, CD8+CD25+Foxp3+, CD4+IL-10+ and CD8+IL-10+Treg subsets; these results have important implications for the development and application of hPMSCs in clinical use (Abstract). The study results demonstrate that Tfr (subset of Foxp3+ Tregs) generation could be promoted by hPMSCs from stimulated PBMC wherein it was reported that programmed death- ligand 1 (PD-L1) promotes Tfr cell formation, and previous results showed that hPMSCs, which express high levels of PD-L1, could negatively regulate T cell functions through suppressing the proliferation of T cells and inducing the generation of IL-10+T cells; additionally it has been demonstrated previously that MSCs induce the generation of CD4+CD25+Foxp3+, CD4+IL-10+ and CD8+IL-10+ Treg subsets and the current study results expand upon these findings and show that these three Treg subsets could be induced by hPMSCs when tested within the same culture system (Page 50, Column 1, Paragraph 2). The generation of CD4+CXCR5+Foxp3+, CD4+IL-10+,and CD8+IL-10+ Treg subsets from activated PBMC were similar, but greater than, that of the CD4+CD25+Foxp3+ and CD8+CD25+Foxp3+Treg subsets with hPMSCs and maximal immunomodulatory effects of hPMSCs pretreated by IFNγ were observed for the generation of the CD4+CXCR5+Foxp3+ Treg subset as compared with that of the other four Treg subsets recorded; the capacity for IFNγ-pretreated hPMSCs to induce the generation of the CD8+IL-10+ Treg subset was maximally enhanced as compared with that of the CD4+CD25+Foxp3+, CD8+CD25+Foxp3+, and CD4+IL-10+Treg subsets and collectively these results indicate that hPMSCs could upregulate the amount of specific Treg cells and, in turn, enhance their immunosuppressive effects (Page 50, Column 2, Second Full Paragraph). Thus, Chang and Yi both support the notion that MSCs (i.e., placental MSCs) facilitate the induction of specific Treg subsets and IFNγ pre-treatment of said MSCs improves the immunosuppressive properties of the MSCs.
Gandolfo provides a reasonable expectation of success for inhibiting and/or reversing kidney failure utilizing CD4+CD25+Foxp3+ Tregs. Gandolfo specifically discloses that T lymphocytes modulate early ischemia-reperfusion injury in the kidney, noting their role during repair is unknown, and the authors studied the role of TCRB+CD4+CD25+Foxp3+ regulatory T cells (Tregs), known to blunt immune responses, in repair after ischemia-reperfusion injury to the kidney (Abstract). Treg depletion starting 1 day after ischemic injury using anti-CD25 antibodies increased renal tubular damage, reduced tubular proliferation at both time points, enhanced infiltrating T lymphocyte cytokine production at 3 days and TNF-α generation by TCRβ+CD4+T cells at 10 days while, in separate mice, infusion of CD4+CD25+ Tregs 1 day after initial injury reduced INF-γ production by TCRβ⁺CD4+ T cells at 3 days, improved repair and reduced cytokine generation at 10 days; the study demonstrates that Tregs infiltrate ischemic-reperfused kidneys during the healing process promoting repair, likely through modulation of pro-inflammatory cytokine production of other T cell subsets and Treg targeting could be a novel therapeutic approach to enhance recovery from ischemic acute kidney injury (Id.; emphasis added). Thus, the direct administration of cultured Tregs (i.e., CD4+CD25+Foxp3+ Tregs) would reasonably be expected to inhibit and/or reverse kidney failure associated with ischemia and renal fibrosis.
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed to isolate and expand autologous Treg cells ex vivo in the presence of MSCs (e.g., placental MSCs), IFN-γ, and IL2 to generate cultured Tregs (i.e., CD4+CD25+Foxp3+ Tregs) and administer said Tregs to a patient having kidney failure associated with ischemia and renal fibrosis. One would have been motivated to combine the teachings of Kim, Kenzel, Chang, and Yi: (1) in order to develop an effective ex vivo method of producing and expanding Treg cell populations effective for treating renal ischemia reperfusion injury, (2) because Kim and Kenzel recognize the need for improving ex vivo expansion Treg cells for therapy, and (3) the cited references teach and recognize that CD4+CXCR5+Foxp3+ Treg cells are effective for therapy. One of ordinary skill in the art would have a reasonable expectation of success because: (1) the cited references teach the established, successful methods of isolating Tregs for renal therapy, inducing and expanding the anti-inflammatory Treg cell phenotype, CD4+CXCR5+Foxp3+, by exposure of Tregs to IL-2 or cultured with IFNγ-treated MSCs, and teach the known benefits of treating kidney failure associated with ischemia and renal fibrosis with CD4+CXCR5+Foxp3+ Treg cells.
In the test of whether it is “obvious to try” there must be:
(1) a finding in the art at the time of filing of the invention that there had been a recognized problem or need in the art;
(2) a finding that there had been a finite number of identified, predictable potential solutions to the recognized need or problem;
(3) a finding that one of ordinary skill in the art could have pursued the known potential solutions with a reasonable expectation of success.
In the instant case: (1) Kim and Kenzel recognize the need for improving ex vivo expansion of autologous Treg cells for renal therapy, and Kim, Kenzel, and Gandolfo recognize the need to treat kidney failure associated with ischemia and renal fibrosis with CD4+CXCR5+Foxp3+ Treg cells;
(2) Kim, Kenzel, Chang, and Yi teach the known process of successfully inducing and expanding CD4+CXCR5+Foxp3+ Treg cells through exposure to IL-2 and IFNγ-treated MSCs; and Kim, Kenzel, and Gandolfo teach the known success of treating kidney failure associated with ischemia and renal fibrosis with CD4+CXCR5+Foxp3+ Treg cells; and
(3) one of skill in the art could have pursued isolating, inducing, and expanding Treg cells ex vivo into an effective population of CD4+CXCR5+Foxp3+ Treg cells and administered them to treat kidney failure associated with ischemia and renal fibrosis with a reasonable expectation of success.
Claims 23-24 are rejected under 35 U.S.C. 103 as being unpatentable over non-patent literature by Kim et. al. (J. Am. Soc. Nephrol., 2013, 24, 1529-1536; herein after referred to as "Kim"), non-patent literature by Landwehr-Kenzel et. al. (Kidney International, 2018, 93, 1452-1464; herein after referred to as "Kenzel"), non-patent literature by Chang et. al. (Stem Cells, 2006, 24(11), 1466-2477; herein after referred to as "Chang"), non-patent literature by Yi et. al. (Cellular Immunology, 2018, 326, 42-51; herein after referred to as "Yi"), and non-patent literature by Gandolfo et. al. (Kidney International, 2009, 76(7), 717-729; previously cited on PTO-892; herein after referred to as “Gandolfo”), as applied to claims 21 and 27 above, and further in view of non-patent literature by Huang et. al. (Scientific Reports, 2015, 5(16565), 1-8; previously cited on PTO-892; herein after referred to as "Huang") and non-patent literature by Levey and Coresh (Lancet, 2012, 379, 165-180; previously cited on PTO-892; herein after referred to as “Levey”).
Kim, Kenzel, Chang, Yi, and Gandolfo teach a method of inhibiting and/or reversing kidney failure associated with ischemia and renal fibrosis comprising the steps of: (a) identifying a patient suffering from kidney failure associated with renal fibrosis; (b) extracting T regulatory cells from a subject; (c) providing mesenchymal stem cells (MSCs) derived from a source selected from the group consisting of: placental tissue, amniotic membrane, umbilical cord tissue, fallopian tube tissue, and subepithelial umbilical cord tissue; (d) contacting said mesenchymal stem cells with interferon gamma in vitro and generating a mesenchymal stem cell conditioned media (MSC-CM); (e) culturing said T regulatory cells with said MSC-CM in vitro in the presence of interleukin 2 in a manner so that the MSC-CM endows onto said T regulatory cells properties capable of preventing, inhibiting and/or reversing kidney failure; and (f) administering said cultured T regulatory cells into said patient.
The combined references do not teach that (i) the patient has a glomerular filtration rate (GFR) below 45 and said GFR is improved after administration of the cultured T regulatory cells; nor (ii) the patient suffers from high levels of proteinuria and said levels are decreased after administration of the cultured T regulatory cells.
Huang teaches that CD4+CD25+ cells (i.e., T regulatory cells) are significantly decreased in multiple myeloma (MM) related renal impairment (RI) patients compared to the controls (P<0.05) wherein CD4+CD25+ cell number was negatively associated with blood urea nitrogen (BUN), supernatant IL-4, serum IL-6, monoclonal immunoglobulin and β2-microglobulin, as well as bone marrow plasma cell percentage and proteinuria while being positively associated with estimated glomerular filtration rate (eGFR) (all P < 0.05); CD4+CD25+ cells gradually decreased as the Clinic Stage increased and the number of CD4+CD25+ cells was reduced in MM related RI patients, and was correlated with disease severity (Abstract). More specifically, Huang teaches that the associations between the numbers of CD4+CD25+ cells and clinical parameters were examined in patients with MM-RI wherein it was observed that CD4+CD25+ cells were negatively correlated with serum blood urea nitrogen (BUN), proteinuria, and uric acid, but positively correlated with eGFR (P < 0.05); these results indicated that CD4+CD25+ cells may represent renal function (Page 4, Paragraph 2; Table 3). CD4+CD25+ cells were negatively correlated with urine protein, supernatant IL-4, serum IL-6, bone marrow plasma cell percentage, as well as blood monoclonal immunoglobulin and β 2-microglobulin (P < 0.05) (Id.). Thus, Huang teaches that increased T regulatory cells in peripheral blood is correlated with higher eGFR and lower proteinuria, i.e., healthier individuals such as the controls in this study have increased T regulatory cell numbers and thus have higher eGFR and lower proteinuria. However, Huang does not explicitly teach a GFR of less than 45 nor proteinuria levels and their direct correlation to kidney disease. This deficiency is remedied by Levey.
Levey teaches that the definition of chronic kidney disease is based on the presence of kidney damage (i.e., albuminuria/proteinuria) or decreased kidney function (i.e., glomerular filtration rate [GFR] <60 mL/min per 1·73 m²) for 3 months or more, irrespective of clinical diagnosis, and that because of the central role of GFR in the pathophysiology of complications, the disease is classified into five stages on the basis of GFR: more than 90 mL/min per 1·73 m² (stage 1), 60–89 mL/min per 1·73 m² (stage 2), 30–59 mL/min per 1·73 m² (stage 3), 15–29 mL/min per 1·73 m² (stage 4), and less than 15 mL/min per 1·73 m² (stage 5) (Page 165, Column 2, Paragraph 1). Risks of both mortality and kidney failure are associated with GFR and concentration of albuminuria; Figure 4 shows US prevalence estimates by eGFR and urinary albumin to creatinine ratio wherein the proportion of participants with chronic kidney disease in the groups at moderate, high, and very high risk (as categorized in figure 2) is about 73%, 18%, and 9%, respectively, representing a prevalence in the general population of about 10%, 2%, and 1%, respectively (Page 168, Column 1, Paragraph 1; Figure 4). Thus, Levey teaches that decreased GFR (e.g., eGFR <45 = stage 3 or higher kidney disease) and increased albuminuria (i.e., proteinuria) present increased risk for chronic kidney disease and renal failure.
It would have been prima facie obvious to one of ordinary skill in the art to apply the method rendered obvious by Kim, Kenzel, Chang, Yi, and Gandolfo to a patient population with kidney disease wherein said population has at least stage 3 disease (i.e., eGFR of <60) and/or wherein the patient has high levels of proteinuria (i.e., albuminuria), because the risks of both mortality and kidney failure are associated with GFR and thus later stage disease as indicated by GFR indicates a patient population in need of treatment as taught by Levey. One of ordinary skill in the art would have a reasonable expectation of success because clinical trends indicate that CD4+CD25+ cells were negatively correlated with serum blood urea nitrogen (BUN), proteinuria, and uric acid, but positively correlated with eGFR (P < 0.05), indicating that CD4+CD25+ Treg cells may represent renal function wherein increased T regulatory cells in peripheral blood is correlated with higher eGFR and lower proteinuria, i.e., healthier individuals such as the controls in this study have increased T regulatory cell numbers and thus have higher eGFR and lower proteinuria, as taught by Huang. Thus, the cultured Treg cells of the obvious method of claim 21 would be expected to, via their immunosuppressive effects, attenuate renal impairment (e.g., in cases of ischemia, fibrosis, and renal failure) which would subsequently improve GFR and reduce proteinuria after cultured Treg administration.
Claim 25 is rejected under 35 U.S.C. 103 as being unpatentable over non-patent literature by Kim et. al. (J. Am. Soc. Nephrol., 2013, 24, 1529-1536; herein after referred to as "Kim"), non-patent literature by Landwehr-Kenzel et. al. (Kidney International, 2018, 93, 1452-1464; herein after referred to as "Kenzel"), non-patent literature by Chang et. al. (Stem Cells, 2006, 24(11), 1466-2477; herein after referred to as "Chang"), non-patent literature by Yi et. al. (Cellular Immunology, 2018, 326, 42-51; herein after referred to as "Yi"), and non-patent literature by Gandolfo et. al. (Kidney International, 2009, 76(7), 717-729; previously cited on PTO-892; herein after referred to as “Gandolfo”) as applied to claims 21 and 27 above, and further in view of Komaki et. al. (Stem Cell Research & Therapy, 2017, 8(219), 1-12; herein after referred to as "Komaki"), non-patent literature by Chen et. al. (Stem Cell Research & Therapy, 2020, 11(91), 1-11; herein after referred to as “Chen”), and non-patent literature by Liang et. al. (Cell Biology, 2018, 23(1), 44-49; herein after referred to as “Liang”).
Kim, Kenzel, Chang, Yi, and Gandolfo teach a method of inhibiting and/or reversing kidney failure associated with ischemia and renal fibrosis comprising the steps of: (a) identifying a patient suffering from kidney failure associated with renal fibrosis; (b) extracting T regulatory cells from a subject; (c) providing mesenchymal stem cells (MSCs) derived from a source selected from the group consisting of: placental tissue, amniotic membrane, umbilical cord tissue, fallopian tube tissue, and subepithelial umbilical cord tissue; (d) contacting said mesenchymal stem cells with interferon gamma in vitro and generating a mesenchymal stem cell conditioned media (MSC-CM); (e) culturing said T regulatory cells with said MSC-CM in vitro in the presence of interleukin 2 in a manner so that the MSC-CM endows onto said T regulatory cells properties capable of preventing, inhibiting and/or reversing kidney failure; and (f) administering said cultured T regulatory cells into said patient.
The combined references do not teach that the MSCs secrete higher levels of hepatocyte growth factor (HGF) than normal.
Komaki evidences a high expression level of HGF in placenta-derived MSC-CM (PlaMSC-CM); expression levels of the angiogenic factor(s) associated with PlaMSC-CM were screened using a growth factor array selected based on previous reports of MSC-CM angiogenic activity using an endothelial tube formation assay; in PlaMSC-CM, the levels of HGF, IGFBP2, IGFBP3, and IGFBP6 were higher than those of other growth factors (Page 5; Figure 2). As such, PlaMSC-CM, as suggested by the obvious method of claim 21, would have high expression of HGF as evidenced by Komaki. Therefore, the invention as a whole was prima facie obvious to one of ordinary skill in the art at the effective filing date of the invention as evidenced by the references.
Chen teaches that mesenchymal stem cells (MSCs) exert immunomodulatory functions by inducing the development and differentiation of naive T cells into T cells with an anti-inflammatory regulatory T cell (Treg) phenotype, wherein the authors’ previous study showed that hepatocyte growth factor (HGF) secreted by MSCs had immunomodulatory effects in the context of lipopolysaccharide (LPS) stimulation; the authors hypothesized that HGF is a key factor in the MSC-mediated regulation of the T helper 17 (Th17) cell/regulatory T (Treg) cell balance (Abstract). The percentage of CD4+CD25+Foxp3+ cells was significantly increased in the CD4+ T cell population, while the percentage of CD4+CD3+RORrt+ cells was significantly decreased after MSC coculture, however the MSC-induced effect was significantly inhibited by the anti-HGF antibody (p < 0.05); MSCs significantly inhibited the CD4+ T cell expression of IL-17 and IL-6 but increased the expression of IL-10 (p < 0.05 or p < 0.01) and these effects were inhibited by the anti-HGF antibody (p < 0.05) (Id). In addition, CD4+ T cells cocultured with MSCs significantly inhibited neutrophil phagocytic and oxidative burst activities (p < 0.05 or p < 0.01), however these MSC induced effects were inhibited by the anti-HGF antibody (p < 0.05); together the data suggests that MSCs induced the conversion of fully differentiated Th17 cells into functional Treg cells and thereby modulated the Th17/Treg cell balance in the CD4+ T cell population, which was partly attributed to HGF secreted by the MSCs (Id.).
Liang teaches that compared to the controls (MSCs alone), MSCs cocultured with IFN-γ expressed significantly higher concentrations of PGE2, HGF and TGF-β1 and the mRNA level of IDO was remarkably increased; human bone marrow-derived MSCs alone notably suppressed T lymphocytes proliferation in vitro and the addition of exogenous IFN-γ did not ablate the immunosuppressive effects of MSCs and the addition of anti-IFN-γ mAb partially restored suppression of T cell proliferation by MSCs (Abstract). Human MSCs constitutively expressed immunosuppressive levels of PGE2, HGF and TGF-β1; the proinflammatory cytokine IFN-γ exhibited synergistic effects with MSCs on immunosuppression, possibly by up-regulating PGE2, HGF and TGF-β1 in MSCs and inducting MSCs expression of IDO, involved in tryptophan catabolism (Id.). Thus, the combination of Komaki, Chen, and Liang supports that MSCs (i) express/secrete HGF; (ii) MSC immunosuppressive/immunomodulatory activities, including Treg stimulation, are mediated by HGF; and (iii) treatment of MSCs with IFN-γ upregulates HGF expression/secretion.
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed that the cultured Tregs of the obvious method of claim 21 would express increased levels of HGF, as supported by the combined teachings of Komaki, Chen, and Liang.
Claim 26 is rejected under 35 U.S.C. 103 as being unpatentable over non-patent literature by Kim et. al. (J. Am. Soc. Nephrol., 2013, 24, 1529-1536; herein after referred to as "Kim"), non-patent literature by Landwehr-Kenzel et. al. (Kidney International, 2018, 93, 1452-1464; herein after referred to as "Kenzel"), non-patent literature by Chang et. al. (Stem Cells, 2006, 24(11), 1466-2477; herein after referred to as "Chang"), non-patent literature by Yi et. al. (Cellular Immunology, 2018, 326, 42-51; herein after referred to as "Yi"), and non-patent literature by Gandolfo et. al. (Kidney International, 2009, 76(7), 717-729; previously cited on PTO-892; herein after referred to as “Gandolfo”) as applied to claims 21 and 27 above, and further in view of non-patent literature by Hirakawa et. al. (Blood, 2015, 126(23), 1-4; previously cited on PTO-892; herein after referred to as “Hirakawa”).
Kim, Kenzel, Chang, Yi, and Gandolfo teach a method of inhibiting and/or reversing kidney failure associated with ischemia and renal fibrosis comprising the steps of: (a) identifying a patient suffering from kidney failure associated with renal fibrosis; (b) extracting T regulatory cells from a subject; (c) providing mesenchymal stem cells (MSCs) derived from a source selected from the group consisting of: placental tissue, amniotic membrane, umbilical cord tissue, fallopian tube tissue, and subepithelial umbilical cord tissue; (d) contacting said mesenchymal stem cells with interferon gamma in vitro and generating a mesenchymal stem cell conditioned media (MSC-CM); (e) culturing said T regulatory cells with said MSC-CM in vitro in the presence of interleukin 2 in a manner so that the MSC-CM endows onto said T regulatory cells properties capable of preventing, inhibiting and/or reversing kidney failure; and (f) administering said cultured T regulatory cells (i.e., CD4+CD25+Foxp3+ Tregs) into said patient.
The combined references do not teach that the cultured Tregs would be CD4+CD25+Foxp3+Helios+ Tregs.
Hirakawa discloses that peripheral blood mononuclear cells (PBMCs) stimulated in vitro with IL-2 (both at low and high concentrations) results in selective activation of Helios+ Treg (Pages 2-3).
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed that the cultured Tregs of the obvious method of claim 21 would be CD4+CD25+Foxp3+Helios+, as supported by Hirakawa in view of Kim, Kenzel, Chang, Yi, and Gandolfo.
Claim 28 is rejected under 35 U.S.C. 103 as being unpatentable over non-patent literature by Kim et. al. (J. Am. Soc. Nephrol., 2013, 24, 1529-1536; herein after referred to as "Kim"), non-patent literature by Landwehr-Kenzel et. al. (Kidney International, 2018, 93, 1452-1464; herein after referred to as "Kenzel"), non-patent literature by Chang et. al. (Stem Cells, 2006, 24(11), 1466-2477; herein after referred to as "Chang"), non-patent literature by Yi et. al. (Cellular Immunology, 2018, 326, 42-51; herein after referred to as "Yi"), and non-patent literature by Gandolfo et. al. (Kidney International, 2009, 76(7), 717-729; previously cited on PTO-892; herein after referred to as “Gandolfo”) as applied to claims 21 and 27 above, and further in view of non-patent literature by Park et. al. (Am. J. Physiol. Renal Physiol., 2017, 313, F984-F996; herein after referred to as “Park”) and non-patent literature by Zhang et. al. (Journal of Reproductive Immunology, 2021, 148, 1-9; herein after referred to as “Zhang”).
Kim, Kenzel, Chang, Yi, and Gandolfo teach a method of inhibiting and/or reversing kidney failure associated with ischemia and renal fibrosis comprising the steps of: (a) identifying a patient suffering from kidney failure associated with renal fibrosis; (b) extracting T regulatory cells from a subject; (c) providing mesenchymal stem cells (MSCs) derived from a source selected from the group consisting of: placental tissue, amniotic membrane, umbilical cord tissue, fallopian tube tissue, and subepithelial umbilical cord tissue; (d) contacting said mesenchymal stem cells with interferon gamma in vitro and generating a mesenchymal stem cell conditioned media (MSC-CM); (e) culturing said T regulatory cells with said MSC-CM in vitro in the presence of interleukin 2 in a manner so that the MSC-CM endows onto said T regulatory cells properties capable of preventing, inhibiting and/or reversing kidney failure; and (f) administering said cultured T regulatory cells (i.e., CD4+CD25+Foxp3+ Tregs) into said patient.
The combined references do not teach that the MSCs are derived from umbilical cord tissue.
Park teaches that human umbilical cord blood (hUCB) derived MSCs were used to compare the effects and mechanisms of early and late MSC therapy in a murine model; after cisplatin injection into C57BL/6 mice, hUCB-MSCs were administered on day 1 (early treatment) or day 3 (late treatment), wherein with early treatment, cisplatin nephrotoxicity was attenuated as evidenced by decreased blood urea nitrogen (BUN) and reduced apoptosis and tubular injury scores on day 3 (Abstract). Early treatment resulted in downregulation of intrarenal monocyte chemotactic protein-1 and IL-6 expression and upregulation of IL-10 and VEGF expression, and flow cytometric analysis showed similar populations of infiltrated immune cells in both groups; however, regulatory T-cell (Treg) infiltration was 2.5-fold higher in the early treatment group and the role of Tregs was confirmed by the blunted effect of early treatment on renal injury after Treg depletion. (Id.). Study data indicates that early hUCB-MSC treatment could attenuate cisplatin-induced nephrotoxicity in the early injury phase by increasing intrarenal Treg infiltration (Page F995, Conclusions).
Zhang teaches that, owning to the notable immunoregulatory potentials, mesenchymal stromal cells (MSCs) and regulatory T cells (Tregs) have been separately reported as promising therapeutic approaches for refractory RSA attributable to certain immune disorders; however, the cross-talk between MSCs and Tregs at the fetal-maternal interface remains poorly understood. The authors revealed that umbilical MSCs could induce expansion of decidual Foxp3+CD4+ T cells with upregulated production of IL-10 and TGF-β and MSCs reinforced the immune suppressive functions of decidual Tregs (dTregs) (Abstract). MSCs-instructed dTregs gained enhanced capacity to suppress Th1 and Th17 related inflammatory responses and in vivo data demonstrated that adoptive transfer of MSCs obviously promoted accumulation of Foxp3+ dTregs in lipopolysaccharide (LPS)-induced mice abortion model and spontaneous abortion model (DBA/2-mated female CBA/J mice) (Id.). To demonstrate whether uMSCs could exert regulatory function on dTregs, the in vitro coculture system of uMSCs and DICs was created and the percentage of Foxp3+CD4+ T cells was analyzed via flow cytometry; as shown in
Fig. 2A, uMSCs treatment substantially drove the expansion of Foxp3 T cell population, which verified the prominent role of uMSCs in promoting the abundance of dTregs at the fetal-maternal interface, and the authors also detected the expression of anti-inflammatory cytokine IL-10 and TGF-β in dFoxp3+CD4+ T cells induced by uMSCs (Page 3, Column 2, Section 3.2). The results in Fig. 2B showed that the production of IL-10 and TGF-β in dTregs was significantly upregulated after uMSCs treatment; all the data indicated that uMSC could induce the augmentation of dTregs and enhance their immunosuppressive properties (Id.). Thus, the combination of Park and Zhang suggest that umbilical tissue derived MSCs also have immunosuppressive/immunoregulatory properties, wherein said umbilical tissue derived MSCs also induce differentiation of Tregs (e.g., CD4+Foxp3+ Tregs) wherein said Tregs subsequently induce immunosuppressive effects and can treat nephrotoxicity injury in early phases.
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed that the MSCs of the obvious method of claim 21 could be umbilical tissue derived MSCs wherein the use of umbilical tissue derived MSCs instead of placental MSCs would reasonably be expect to produce the same results (i.e. produce Tregs capable of exerting immunosuppressive effects), as supported by the combined teachings of Park and Zhang in view of Kim, Kenzel, Chang, Yi, and Gandolfo.
Response to Arguments
Applicant’s arguments presented in Remarks (12/02/2025) with respect to currently pending claims 21 and 23-28 have been considered but are moot because the new ground(s) of rejection do not rely on any reference applied in the prior rejection of record for any teaching or matter specifically challenged in the argument.
Conclusion
Claims 21 and 23-28 are pending. Claims 21 and 23-28 are rejected. No claims are allowed.
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/ALYSSA RAE STONEBRAKER/Examiner, Art Unit 1642
/Laura B Goddard/Primary Examiner, Art Unit 1642