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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 11/19/2025 has been entered.
Claim Status
Claims1, 3, and 6-8 are amended.
Claims 1-8 are under examination.
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.
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.
Claims 1-8 are rejected under 35 U.S.C. 103 as being unpatentable over Zhou et al (Nature Protocols, 2012) in view of Yoon et al ( US 2015/0307840 A1), Shou et al ( CN 106282238 A, published 01/04/2017), Shou et al (CN110042125A, published 07/23/2017), and Zhang et al (WO 2008/153685 A2), as evidenced by Lonza (Clonetics™ Endothelial Cell System, Lonza),Sigma-Aldrich product information sheet, Lonza product information sheets ( REGM™ SingleQuots Kit and REGM™ Bullet Kit). CN 106282238A and CN110042125A are in the Chinese language. A machine translation is provided herewith. Citations are made to the machine translation.
Regarding claim 1, Zhou et al disclose a method for generating induced pluripotent stem cells (iPSCs) by reprogramming cells isolated from urine samples. (See abstract). The method of Zhou et al includes concentrating urine cells isolated from a subject by centrifugating the collected urine at 400xg for 10 minutes at room temperature. This reads on step i of instant claim. ( See “ isolation of urine cells” under the Procedure section, step 3, page 2084). The method of Zhou et al also includes a step for expanding (e.g. replicating) the urine cells prior to transfection. The first step in the expanding process is to culture the concentrated urine cells in primary medium for three days. The primary medium of Zhou comprises of DMEM/high glucose and Ham’s F12 nutrient mix (1:1), supplemented with 10% (vol/vol) FBS, 100 U/ml of penicillin, 100 µg/ml of streptomycin, 2.5 µg/ml amphotericin B, and REGM SingleQuot kit supplements, wherein the REGM SingleQuot kit components include EGF, Hydrocortisone, and Epinephrin as evidenced by the Lonza product information sheet. See Zhou et al ( “ Procedure” section “ Urinary cell expansion”, step 11, page 2084, and for primary medium composition see “ Reagent Setup” section “ Primary medium”, page 2083) and Lonza product information sheet (REGM™ SingleQuots Kit Content, page 2-3). The second step in the expansion process, further includes culturing cells in proliferation medium for 9-12 days. The proliferation medium comprises of basal renal epithelial growth medium supplemented with RGEM BulletKit components, wherein the BulletKit components include serum, hydrocortisone, EGF, and epinephrin as evidenced by Lonza product information sheet . See Zhou et al (Procedure” section “ Urinary cell expansion”, step 12, page 2085, and for proliferation medium composition See “ Reagent Setup” section “ RE proliferation medium”, page 2083) and Lonza product information sheet ( REGM™ Bullet Kit Content, page 3 ). This reads on steps ii of instant claim.
Collectively, Zhou et al teach steps (i) and (ii) of instant claim, which are carried to concentrate and replicate the isolated urine cells. Zhou et al, however, do not teach steps iii, iv, and v of instant claim.
Yoon et al teach a method for producing endothelial-like cells. The method involves exposing fibroblasts to a recombinant vector encoding the transcription factor ETV2. (See paragraph [0013]). Yoon et al’s method also involves the step of culturing the transformed fibroblasts in a first medium containing DMEM supplemented with 10% fetal bovine serum for 7 days providing endothelial cell lineage. (See paragraph [0084]). This reads on step iii of instant claim. Yoon et al teach after 7 days, the cells were transferred into a second growth medium consisting of EGM-2 medium containing DOX and the cells were further cultured on collagen-coated plates, wherein the EGM-2 is an endothelial growth medium. (See paragraph [0084]). Yoon et al do not explicitly recite the components claimed in step v, however, it is well known that the EGM2 medium contains these components as evidenced by Lonza standard protocols. ( See page 7 , 2nd column “ IX. Preparation of culture media” step 4). This reads on step v. It should be noted that the second growth medium used by Yoon et al is a low serum culture medium designated for the growth and maintenance of the produced endothelial cells. Therefore, an ordinary skill in the art at the time the invention was filed would be motivated to combine the teachings of Zhou and Yoon to use two different culturing medium while reprogramming urine progenitor cells (UPCs) into endothelial cells. Because Zhou et al teach that the first culturing medium can be used to increase UPCs proliferation, providing enough cell numbers for the retroviral infection and so ensuring higher endothelial cell production. Yoon et al teach that after 7 days of infection, the cells should be switched to a second culturing medium designated for the growth and maintenance of endothelial cells. Thus, one would be motivated to combine the teachings of Yoon and Zhou to culture the urine derived cells into a first medium, as taught by Zhou et al, as doing so would provide a larger number of UPCs that can be used in research or clinical applications. One would also be motivated by the teachings of Yoon et al to switch the cells after reprogramming into a second growth medium, as doing so would provide the reprogrammed cells with the factors required for endothelial cell growth and maintenance.
It is noted that neither Zhou et al nor Yoon et al suggest or teach using UPCs to produce endothelial cells.
However, according to Shou et al (2017) “Studies have shown that in vitro transfection of specific transcription factors into cells can differentiate terminally differentiated cells into functional blood vessels, nerves and other cells”. Shou et al go on to demonstrate the possibility of using a recombinant vector encoding the transcription factor ETV2 to reprogram muscle progenitor cells (e.g. HskMM) into endothelial-like cells. (See examples 3-4 on pages 12-13. Additionally, Shou et al teach that the transcription factor ETV2 may also be used to produce endothelial-like cells from other cell types, including human myoblast, a fibroblast, an umbilical cord-derived mesenchymal stem cell, an adipose stem cell, or a glioma cell. ( See 3rd paragraph on page 4).
Moreover, Shou et al (2019) demonstrate using a recombinant vector encoding ETV2 to reprogram adipose derived stem cells (ASCs) into endothelial-like cells. Shou et al, teach that using the ETV2 expressing vector, the ASCs can transdifferentiate into endothelial-like cells at a high efficiency.(See examples 3-4 on pages 11-12).
On the other hand, Zhang et al teach methods for producing a culture of urine progenitor cells (UPCs ), including providing a urine sample; centrifugation of the collected urine sample; and then isolating urine progenitor cells from said urine sample.(See example 1 on page 17). Zhang et al teach that the UPCs can be differentiated into two or more lineages selected from the group consisting of: urothelium, smooth muscle, endothelium, or interstitial cells. ( See the 5th paragraph on page 3).
Taken together, the teachings of Yoon, Shou (2017), and (2019) suggest a potent determinant function for ETV2 in converting different cells types into endothelial cells. Zhang teach that UPCs have the potential to be differentiated into endothelial cells.
Therefore, claim 1 would have been obvious to one of ordinary skill in the art, as there was some teaching, suggestion, or motivation in the prior art that would have led one of ordinary skill to modify the prior art reference or to combine prior art reference teachings to arrive at the claimed invention. Zhou et al suggest a culturing medium that is suitable for the propagation and expansion of urine-derived cells, but fails to teach producing endothelial cells from UPCs. Yoon et al demonstrate how to reprogram fibroblast into endothelial-like cells using a vector encoding the transcription factor ETV2. Shou et al (2017) and (2019) demonstrate that a vector encoding ETV2 can be used to reprogram different cell types into endothelial cell lineages. Zhang et al teach that UPCs have the potential to be differentiated into endothelial like cells, smooth muscle cells, or urothelium. Therefore, one with ordinary skill in the art who had reviewed Zhou, could have come across Yoon, Shou (2017) /(2019), and Zhang and immediately noticed the strong possibility of using a vector encoding ETV2, as taught by Yoon, Shou (2017) and (2019), to produce endothelial-like cells using UPCs. One would have been motivated to substitute the cells of Yoon with UPCs, because UPCs are easily isolated from human urine. Also, using human urine cells would eliminate the pain associated with sampling source cells from the skin to isolate dermal fibroblasts. There is a reasonable expectation of success that a vector encoding ETV2 can be used to produce endothelial-like cells from UPCs.
The method of Zhou and Yoon do not teach the transdifferentiating of urine derived progenitor cells (UPCs) into smooth muscle cells. However, the combined teachings of Zhou and Yoon include the same steps as the instant claim, therefore transdifferentiating UPCs would result in the inherent production of smooth muscle cells.
Furthermore, with regard to the limitations directed to generating endothelial cells and smooth muscle cells expressing the specific markers recited in instant claim, these limitations are directed to results that flow from performing the steps of the method, and do not particularly provide patentable weight in determining patentability of the claimed methods. In addition, as the combined teachings of Zhou et al and Yoon et al teach the claimed method steps, the results are expected the same. Regardless, Yoon et al teach that the produced endothelial cells express CDH5, PECAM1, and KDR. (See Fig.1D).
Regarding claim 2, following the discussion above, Zhou in view of Yoon render obvious claim 1. Zhou in view of Yoon teach using animal serum rather than human serum. However, it would have been prima facie obvious to one with ordinary skill in the art to substitute the source of the serum from animal to human source in steps ii and v (d) of claim 1, as doing so would make the produced endothelial and smooth muscle like cells pharmaceutically suitable for human transplantation because one would expect such a substitution would not elicit any immune response due to antigens produced by the use of fetal bovine serum.
Regarding claim 3 and 4, following the discussion of claim 1, Zhou teach a culturing medium for urine derived cells that significantly propagate cell numbers required for research or clinical applications. Yoon et al teach the use of lentiviral vector encoding for the ETV2 is sufficient to successfully transdifferentiate progenitor cells into endothelial like cells. Shou et al (2017) and (2019) demonstrate that a vector encoding ETV2 can be used to reprogram different cell types into endothelial cell lineages. Zhang et al teach that UPCs have the potential to be differentiated into endothelial cells. Therefore, it would have been prima facie obvious to one with ordinary skill in the art at the time the invention was filed to substitute the cells of Yoon with urine derived cells of Zhang et al to produce endothelial-like cells and culture them in a first medium that contains the components suggested by Zhou et al. One would have been motivated to do so because the urine derived cells are easily isolated from human urine. Also, using human urine cells would eliminate the pain associated with sampling source cells from the skin to isolate dermal fibroblasts.
Regarding claim 5, following the discussion of claim 1, both Zhou and Yoon disclose the use of DMEM as the base medium of the first growth medium. But they do not explicitly recite its components. However, it is well known in the art that DMEM contains glucose, amino acids, vitamins, glutamine and sodium pyruvate as evidenced by Sigma-Aldrich product information sheet. (See table page 1-3).
Claim 6-8 are rejected under 35 U.S.C. 103 as being unpatentable over Zhou et al (Nature Protocols, 2012) in view of Yoon et al, Shou et al (2017), Shou et al (2019), and Zhang et al , as applied to claims 1-5 above, and further in view of Ganesan et al (Tissue Engineering, 2017) and Zhang et al (WO 2008/153685 A2).
Regarding claim 6, following the discussion above, Zhou et al in view of Yoon render obvious the production of endothelial and smooth muscle like vasculature tissue from urine derived cells. However Zhou and Yoon do not teach folding the endothelial and smooth muscle like vasculature into a three-dimensional structure. Ganesan et al disclose a method of folding human smooth muscle cells derived from umbilical artery and endothelial cells derived from human umbilical vein into three-dimensional sandwich coculture using µ-slides. (See abstract).
Therefore, it would have been prima facie obvious to one with ordinary skill in the art at the time the invention was filed to combine the teachings of Zhou, Yoon, and Ganesan to fold the endothelial and smooth muscle like vascular tissue into a three-dimensional structure such as a blood vessel. Zhou and Yoon teach methods of generating endothelial and smooth muscle cells from urine derived cells. Ganesan et al teach a method of folding the two types of cells into three-dimensional structure. Thus, one would have been motivated to generate the endothelial and smooth muscle like vascular tissue, as disclosed by Zhou and Yoon, and fold them into a three-dimensional structure, as disclosed by Ganesan, because it would have enabled tissue such as engineered blood vessels for implantation into a subject. Combining prior art elements according to known methods to yield predictable results. See MPEP 2143 (I)(A).
Regarding claim 7 and 8, following the discussion of claim 1 above, Zhou in view of Yoon and Ganesan render obvious the production of endothelial and smooth muscle cells from urine derived cells and folding them into three-dimensional structure. Yoon et al further disclose “the use of endothelial cells for treating or preventing a skin condition, vascular condition, heart disease, etc. comprising administering an effective amount of cells to a subject in need for treatment”. ( See paragraph [0025] and claim 8). However, Zhou, Yoon and Ganesan do not teach contacting the cells with a vein, artery, or capillary, etc. Zhang et al teach methods of isolating urine progenitor cell and differentiating them into urothelium, smooth muscle, and endothelium. Zhang et al also disclose methods of treating a subject in need by providing a bladder tissue substrate that includes differentiated urine cells and transplanting the substrate into the patient. (See abstract) . Furthermore, Zhang et al disclose "In further embodiments, formulations of the invention include those for parenteral administration (e.g., administration is carried out intravascularly, either by simple injection, or by injection through a catheter positioned in a suitable blood vessel, such as renal artery". (See page 15, lines 11-17).
Therefore, it would have been prima facie obvious to one with ordinary skill in the art at the time the invention was filed to combine the teachings of Zhou, Yoon, Ganesan, and Zhang to generate the endothelial and smooth muscle like vascular tissue, as disclosed by Zhou and Yoon, and fold them into a three-dimensional structure, as disclosed by Ganesan, and then implanting them into a subject in need by contacting them with an artery, as disclosed by Zhang et al, because using urine derived cells as autologous endothelial and smooth muscle sources for vessel and tissue engineering strategies can yield a sufficient number of cells via a noninvasive, simple, and low-cost method suitable for rapid clinical translation. Combining prior art elements according to known methods to yield predictable results. See MPEP 2143 (I)(A).
Response to Arguments
Applicant's arguments filed 08/19/2025 have been fully considered but they are not
persuasive.
Applicants argue that the cited references fail to disclose or suggest each and every element of the claims. Specifically, Applicants argue that none of the cited references either alone or in combination disclose or suggest producing vascular tissue that includes both endothelial and smooth muscle cells.
Examiner’s Response to Traversal: Applicant’s arguments have been carefully considered but are not found persuasive. This is because, as noted in the rejection above, Zhou et al in view of Yoon, Shou (2017), Shou (2019), and Zhang et al render obvious the method of reprograming UPCs into endothelial cells as recited in claim 1. While none of the cited references recognize the simultaneous production of smooth muscle cells, the production of these cells is considered to be an inherent property because it naturally flows from performing the method. In this case, prior arts render obvious the active steps of the method, which involves contacting UPCs with a vector encoding ETV2, to reprogram them into endothelial lineages. Applicants are reminded that the recitation of an additional advantage associated with doing what the prior art already suggests does not lend patentability to otherwise unpatentable invention.
As per the MPEP, “ The fact that the inventor has recognized another advantage which would flow naturally from following the suggestion of the prior art cannot be the basis for patentability when the differences would otherwise be obvious. See Ex parte Obiaya, 227 USPQ 58, 60 (Bd. Pat. App. & Inter. 1985).
Applicants also argue that exposing urine-derived cells to ETV2 does not always results in the formation of vascular tissue containing both endothelial and smooth muscle cells, hence it cannot be regarded as an inherent result. Rather, the Applicants argue that this is an unexpected result that demonstrate an unpredictable technical achievement. The Applicants in this argument relied on Le Bras et al’s findings, which demonstrated that ETV2 is specifically involved in endothelial cell differentiation and inhibits smooth muscle cell differentiation.
Examiner’s Response to Traversal: Applicant’s arguments have been carefully considered but are not found persuasive. This is because while Le Bras et al demonstrate that overexpressing ETV2 in Sca1+ adventitial cells leads to endothelial differentiation and decreased expression in muscle-related genes, such as αSMA; however this reduction does not imply a complete halt in the production of smooth muscle cells.(See Fig.4B-D). In fact, the results presented in Figure 4 clearly support that, while the production of smooth muscle cells is significantly decreased; it is not fully abolished, supporting the inherent production of smooth muscle cells by the method steps taught by prior arts. Furthermore, prior art by Zhang et al (of records) teaches methods for differentiating urine-derived cells into two or more lineages, such as urothelium, endothelial, smooth muscle, or interstitial cells.(See “ summary of invention” page 3). Zhang et al teach that urine-derived cells can be transdifferentiated into smooth muscle-like cells by culturing them in DMEM containing 10% fetal bovine serum. (See page 8 ,2nd paragraph). It is noted that Yoon’s method includes culturing the transfected urine-derived cells in the first growth medium, comprising DMEM supplemented with 10% fetal bovine serum (FBS), for 7 days.(See paragraph [0084]). Thus, one would expect the inherent production of smooth muscle cell by the method of Yoon et al, as the presence of DMEM and 10% FBS would be effective at transdifferentiating some cells into smooth muscle cells.
Applicants further argue unexpected results by providing additional evidence that the claimed method steps produce vascular tissue containing reprogrammed endothelial and smooth muscle like cells that can fold and remain for more than 3 months in vivo.
Examiner’s Response to Traversal: Applicant’s arguments have been carefully considered but are not found persuasive. This is because, while the specification indicates that the vascular tissue is retained for 3 months, the specification does not provide any comparison that show how these results differ from the other types of tissue produced by prior art. The specification must include an objective comparison supporting that the claimed method steps produce unexpected results and not an inherent property. Applicants are advised to provide a clear evidence that the method of the cited prior art does not possess a critical characteristic that is possessed by the claimed method, doing so would advance prosecution and might permit allowance of applicants' claimed method. Because the claimed method does not recite or present supportive evidence that would account for such a difference (for example, an objective comparison demonstrating that the vascular tissue produced by the claimed method steps is indeed retained in vivo for a longer period of time than the tissue produced by prior art method steps), the combined teachings of prior arts would be presumed to inherently produce vascular tissue containing both endothelial and smooth muscle cells.
For at least these reasons, applicants' arguments are not persuasive
Conclusion
No claim is allowed.
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/FATIMAH KHALAF MATALKAH/Examiner, Art Unit 1638
/Tracy Vivlemore/Supervisory Primary Examiner, Art Unit 1638