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 .
This action is in response to the papers filed on 09/05/2023. Claims 1, 2, 4, 14, 16, 27-31, 40-42, 45, 47-52 are currently pending as per claims filed on 09/05/2023. Claims 3, 5-13, 15, 17-26, 32-39, 43-44, and 46 are cancelled as per claims filed on 09/05/2023.
Therefore, claims 1, 2, 4, 14, 16, 27-31, 40-42, 45, 47-52 are examine in their merits to which the following grounds of rejection are applicable.
Priority
The instant application is a continuation of 17/902,255 filed 09/02/2022 (abandoned) which is a continuation of PCT/US21/20874 filed on 03/04/2021 which claims benefit of provisional 62/985,876 filed on 03/05/2020.
Thus, the earliest possible priority for the instant application is 03/05/2020.
Claim Objections
Claim 48 is objected to because abbreviations such as LCK, FYN, and ZAP-70 should be spelled out at the first encounter in the claims. Appropriate action is required.
Claim 49 is objected to because abbreviations such as NKG2D. Appropriate action is required.
Claim 52 is objected to because abbreviations such as TIRAP, TRIF, and TRAM should be spelled out at the first encounter in the claims. 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.
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.
Claim 1 is vague and indefinite in the recitation of “expressing a system in a cell”. It is unknown how the system is expressed in the cell. The system could be expressed at the transcript level or protein level. The specification describes viral and non-viral delivery vehicles and targeting moieties that may enhance targeting and binding of the delivery vehicle to the cell. Yet, it is unclear how the system is ultimately “expressed in a cell”. As such the metes and bounds of the claim are indefinite.
Claim 1 is vague and indefinite in the recitation of “contacting the cell with the ligand”. It is unknown whether the contacting of the ligand to the cell occurs intracellularly or extracellularly. Similarly, it is unknown how the ligand would bind to the ligand binding domain of the receptor (i.e. is the binding domain of the receptor intracellular and, if so, how would ligand reach the intracellular binding domain?). As such the metes and bounds of the claim are indefinite.
Claim 1, 2, 4, 16, 27-31 are vague and indefinite in the recitation of “activated” and “activable”. It is unknown how the polypeptides can be activated or are activable. This could be achieved by a conformation change, a chemical modification, it is cleaved, or it binds with another molecule such that it has an altered functionality. As such the metes and bounds of the claim are indefinite.
Claims 1 and 27 are vague and indefinite in the recitation of “…capable of…”, since this phrase refers to a latent ability, and it is unknown whether the ability is expressed or observed in the invention. As such the metes and bounds of the claim are indefinite.
Note, it has been held that the recitation that an element is “capable of” performing a function is not a positive limitation, but only requires the ability to so perform. It does not constitute a limitation in any patentable sense. In re Hutchinson, 69 USPQ 138.
Claim 14 further limits claim 1 which requires “one of the first and second chimeric polypeptides [e.g, “the first chimeric polypeptide] comprises a gene modulating polypeptide (GMP) comprising an actuator moiety linked to a cleavage recognition site”. Claim 14 requires “the first chimeric polypeptide comprises the cleavage moiety”. It is unclear if the cleavage moiety is placed along with the actuator moiety linked to a cleavage recognition site. As such the metes and bounds of the claim are indefinite.
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.
The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph:
Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], 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.
Claim 52 rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends. Claim 52 recites wherein the first adaptor moiety or the second adaptor moiety comprises a list of species, however Claim 50 identically recites wherein the first adaptor or the second adaptor moiety comprises a list of species, thus claim 52 does not limit claim 50. Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements.
Claim Interpretation
The terms "activated" and "activatable" used in the claims are not defined in the specification. It is interpreted that a polypeptide is "activated" or “activatable” if it has an induced expression, a conformation change, a chemical modification, it is cleaved, or it binds with another molecule such that it has an altered functionality.
The term "adaptor" is not defined in the specification and has no established meaning in the art. It is interpreted that any chemical moiety may constitute an "adaptor".
Claim Rejections - 35 USC § 112 – Scope of Enablement
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claim 1, 2, 4, 14, 16, 27, 28-31, 40-42, 45, 47-52 rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for
a system for regulating expression of a target polynucleotide in a human primary T cell wherein the T cell comprises:
at a TCR (as the ligand binding domain);
and the system comprising first chimeric polypeptide with a first Zeta Chain of T Cell Receptor Associated Protein Kinase 70 (ZAP70) as the adaptor moiety that is coupled to a Tobacco Etch Virus (TEV) protease cleavage moiety, optionally operatively coupled to a reporter (e.g., truncated rat nerve growth factor receptor (tNGFR));
a second chimeric polypeptide with Linker for Activation of T Cells (LAT) that is coupled to a Cas enzyme or a modification thereof (e.g., dCas-KRAB; actuator moiety), optionally operatively coupled to a reporter (e.g., Q8),
where upon activation of the TCR, the first chimeric polypeptide binds to an intracellular portion of the TCR (ZAP70 domain binds to the intracellular portion of the TCR), and the second chimeric polypeptide binds to the first chimeric polypeptide (e.g., via binding between ZAP70 and LAT), to promote TEV (cleavage moiety) to release the actuator moiety from the second chimeric polypeptide,
does not reasonably provide enablement for a genus of cells or any origin besides a human primary T cell comprising the TLR (or any other endogenous receptor besides TCR), a genus of adaptor moieties besides ZAP70, GRB2, and LAT, any cleavable moiety actuator moiety besides Cas9 and any actuator moiety besides sgRNA that is release from the from the GMP and capable of regulating the expression of the target polynucleotide in any cell. The claim(s) contains subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention.
The factors to be considered in determining whether undue experimentation is required are summarized in In re Wands, 858 F.2d 731, 737, 8 U.S.P.Q.2d 1400, 1404 (Fed. Cir. 1988) (a) the breadth of the claims; (b) the nature of the invention; (c) the state of the prior art; (d) the level of one of ordinary skill; (e) the level of predictability in the art; (f) the amount of direction provided by the inventor; (g) the existence of working examples; and (h) the quantity of experimentation needed to make or use the invention based on the content of the disclosure. While all of these factors are considered, a sufficient number are discussed below so as to create a prima facie case.
the breadth of the claims; the nature of the invention:
The claims are directed to a method of using a system for regulating expression of a target polynucleotide in a cell, comprising: a first chimeric polypeptide and a second chimeric polypeptide, each with adaptor moieties that are activable (claim 2) and wherein the cell comprises a receptor having a ligand binding domain specific for a ligand, wherein the first and second chimeric polypeptides are activatable upon contacting of the cell by the ligand that binds specifically the ligand binding domain of the endogenous receptor.
The claims are broadly but reasonable interpreted as comprising a genus of cells, a genus of receptors having a ligand binding domain specific for a ligand, a genus of adaptor moieties, a genus of cleavage recognition sites and a genus of cleavage moieties capable of cleaving the cleavage recognition sites of the gene modulating polypeptide (GMP) comprising any and all actuator moieties. The Specification only provides support for T cells comprising a T cell receptor, ZAP70, GRB2 and LAT as adaptor moieties, Cas9 as actuator moiety, and an sgRNA against the polynucleotide sequence PD1 or no sgRNA. The actuator is released from the GMP and capable of regulating the expression of the target polynucleotide in the cell.
the amount of direction provided by the inventor; the existence of working examples:
The specification does not disclose a reduction to practice of any endogenous receptor (i.e. nuclear receptor, cytoplasmic receptor, etc) besides a TCR, any adaptor moiety besides ZAP70, GRB2, and LAT, and any actuator moiety besides Cas9. The specification does disclose the receptor is a T cell receptor (TCR) comprising a transmembrane domain and intracellular signaling domain which is activated intracellularly as recited from claim 45 (page 104-106, para 00270-00276) and provides data to support use of at least an intracellular portion of a T cell receptor via an adaptor moiety adaptor moieties ZAP70, GRB2, and LAT, wherein the actuator moiety Cas9 is coupled to LAT and cleavage moiety is coupled to ZAP70 or GRB2 (Fig 7 data shown below for ZAP70 and GRB2).
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Moreover, the working examples in the specification use specific lentivirus constructs and only human primary T cells. Therefore, it is unknown if 1) a portion of a receptor being from a TLR or any other endogenous receptor (i.e. nuclear and cytoplasmic) besides TCR comprising an extracellular binding domain, transmembrane and intracellular domain, 2) any actuator that is release from the GMP and capable of regulating the expression of the target polynucleotide in the cell other than Cas9 and 3) any other adaptor besides LAT coupled to the Cas9 actuator moiety and ZAP70 or GRB2 coupled to the cleavage moiety, wherein ZAP70 and LAT bind, would exhibit the same phenotypic effect of activating gene regulation system. Outside of this scope of enablement, there is no minimal or maximal phenotypic result which would necessarily tell a skilled artisan the system was able to regulate expression of a target polynucleotide in a cell using at least of portion of a TLR, any adaptor protein besides ZAP70 and GRB2, and any actuator besides Cas9.
the state of the prior art; the level of predictability in the art:
The prior art of Chung (Science. 2019) teaches a method of inducing apoptosis in cancer cells by regulating target gene expression using a synthetic cell system dependent on ErbB receptor signaling (page 1, para 1-3; page 6, col 2, para 1-4). More specifically, the constitutive phosphorylation of the receptor (ErbB is a receptor tyrosine kinase (RTK)) which then bind to phosphotyrosine-binding (PTB) and SH2 domains (i.e. the adaptor moieties) of the first and second polypeptides, are responsible for downstream signaling. This signaling leads to the cleavage of a substrate to permit its gene-regulating activity. However, Chung does not predict that a receptor other than a RTK that is constitutively activated and not contacting the cell with a ligand that binds specifically the ligand binding domain would lead to the ability to activate the synthetic gene regulation cell system.
The prior art of Kipniss (Nature Comm. 2017) teaches a C-terminus of a G-protein coupled receptor (GPCR) is fused with a dCas9 effector which contains a cleavage site. Moreover, an adaptor protein, which interacts during GPCR activation, is fused to a protease (TEVp). To release the dCas9 effector, thus enabling gene expression regulation of a target polynucleotide, the protease cleaves the cleavage sequence placed at the N-terminus of dCas9 (Results, page 2, para 1). However, Kipniss does not predict that a receptor other than a GPCR would lead to the ability to activate the synthetic gene regulation cell system.
Fernandez-Aguilar teaches that ZAP70 phosphorylates the membrane adaptor LAT (Linker for the activation of T cells), leading to the formation of the LAT signalosome, including the SLP-76 cytosolic adaptor and phospholipase C-γ1 (PLC-γ1). Although these activatory signaling pathways have been intensely examined,
many questions remain to be elucidated with regard to the regulatory mechanisms, which prevent excessive immune responses, potentially leading to uncontrolled and potentially harmful immune responses (page 2-3, para 3). Hence, although there is evidence that ZAP70 can interact with other adaptor moieties such as SLP76 and PLC-γ as listed in claim 28, there is less predictability in binding of ZAP70 to other adaptor moieties.
the quantity of experimentation needed to make or use the invention based on the content of the disclosure:
The skilled artisan would be required to perform under levels of experimentation in order to practice the claimed invention. The instant specification does not reduce to practice the claimed invention; the instant specification does not provide guidance on how to reasonably predict if at least a portion of Toll-like receptor (TLR), let alone one the species from the recited group, would permit the system to regulate expression of a target polynucleotide in the same manner (i.e. using chimeric polypeptides with adaptor moieties which are activatable) as using a T-cell receptor (TCR).
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The specification discloses 3 lentivirus constructs that are used for transduction into human primary T cells: Lentivirus #1 construct encoding EFla-LAT-tcs-dCas9-KRAB-P2A-Q8, Lentivirus #2 construct encoding mU6-PD1 gRNA/EF1a-Adaptor-TEV- P2A-tNGFR, and Lentivirus #3 construct encoding EF1a-Adaptor-TEV-P2A-tNGFR, without encoding any sgRNA. Moreover, activation of the human primary T cells is activated by an antigen of TCR (e.g., OKT3/CD28). Flow cytometry data in the specification indicates transduction of aforementioned lentiviruses into engineered primary T cells and positive cells are sorted for further expansion. Moreover, data in the specification indicate reduced gene expression of PD1 (gene of interest as targeted by sgRNA) in engineered primary T cells as indication of successful expression of the gene regulating system in the cell. In other words, the effects elicited by the system are demonstrated by reduction in expression of the PD1 gene in primary human T cells.
Moreover, the specification does not disclose how one would utilize an adaptor protein, as shown in Figure 3B and 3C. The constructs shown in Figure 4 disclose the placement of an adaptor protein (ZAP70, GRB2, and LAT) within the constructs, but do not demonstrate how an adaptor protein outside of the construct (i.e. Fig 3B and 3C) would be used.
Thus, the skilled artisan would be forced to determine if any portion of a TLR, use of other adaptor proteins besides LAT, ZAP70, and GRB2, and any actuator besides Cas9 would be necessary and sufficient to elicit the effects of gene regulation in with the system. The currently disclosed invention only describes a system where PD1 gene reduction is achieved in human primary T cells. .
the level of one of ordinary skill:
The level of one of ordinary skill is a PhD holder.
Conclusion
When all of the Wands factors are considered together, they establish a prima facie case that the specification is not enabling for the claims. The Specification only provides 3 lentivirus constructs that are used for transduction into human primary T cells wherein the constructs are transfected into said cells and cause decreased PD1 expression upon activation of a ligand-bound TCR. More specifically, the activation of said receptor causes the release of a Cas9 such that it can be guided by the sgRNA to the PD1 gene and decrease its expression. Moreover, the prior art highlights the unpredictability of using any other receptor besides an RTK or a GPCR (as shown by Chung and Kipniss, respectively) for a cellular gene expression regulation system and that there is incomplete knowledge of how the adaptor proteins interact as they could cause a harmful immune response (as shown by Fernandez-Aguilar).
While a lack of a working embodiment cannot be a sole factor in determining enablement, the lack of any working examples, in light of the unpredictable nature of the art and the lack of direction applicants present, provides additional weight to the lack of enablement in consideration of the Wands factors as a whole. Thus, one of ordinary skill in the art would not have had a reasonable expectation of success in making or using the claimed invention.
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.
To the extent that the claims broadly embrace any cell, any receptor having a ligand binding domain specific for a ligand, any adaptor moieties, any cleavage recognition sites and any cleavage moieties, the following rejection applies.
Claim(s) 1, 2, 4, 14, 16, 27-31, 40-42, 45, 47-50, 51 and 52 is/are rejected under 35 U.S.C. 103 as being unpatentable over Chung et al (US Patent Application No. 2019/0256833 A1) and further in view of Qi et al (US Patent Application No. 2020/0071729 A1) and Wang et al (WO 2019/014390 A1).
Regarding claims 1 and 27, Chung teaches a method and a system for targeted treatment of cancer associated with hyperactivity of a receptor tyrosine kinase, wherein the method regulates expression of a target polynucleotide by expressing a system in a cell. Said method comprising:
a first fusion protein comprising a protease connected to a phosphotyrosine binding (PTB) domain capable of binding to a constitutively phosphorylated tyrosine residue on the receptor tyrosine kinase, reading on a chimeric polypeptide comprising a cleavage moiety (protease) and an adaptor protein (e.g, to a phosphotyrosine binding (PTB) domain) of claim 1; and
b) a second fusion protein for administration comprising an SH2 domain connected to i) a substrate comprising a cleavage site recognized by the protease (Summary, para 0007, page 1) and ii) an anti-cancer therapeutic agent (i.e. the anti-cancer therapeutic agent can comprise a complex of a catalytically inactive Cas9 with a guide RNA for activating or repressing expression of a gene of interest; para 0019) wherein cleavage of the substrate at the cleavage site by the protease of the first fusion protein releases the anti-cancer therapeutic agent from the second fusion protein (Summary, para 0007, page 1), reading on a chimeric polypeptide comprising a gene modulating polypeptide comprising an actuator moiety (e.g, a catalytically inactive Cas9 with a guide RNA for activating or repressing expression of a gene of interest ) linked to a cleavage recognition site, which actuator moiety is capable of regulating the expression of the target polynucleotide in the cell of claim 1.
However, Chung does not teach a receptor having a ligand binding domain, contacting the cell with the ligand that binds specifically the ligand binding domain, as the first and second chimeric polypeptide are activated as Chung’s system gets activated by hyperactivity of the ErbB receptor tyrosine kinase.
Qi teaches “a system for regulating expression of a target polynucleotide in a cell. The system comprises: a chimeric receptor polypeptide that undergoes a receptor modification including a conformational change or chemical modification upon binding to a ligand for the GPCR” (Summary, para 0005, page 1; Figure 28A & B show binding of ligand to receptor on the cell shown below).
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Moreover, Qi teaches “in response to antigen binding, the receptor is modified by phosphorylation in the intracellular region of the receptor (FIG. 8B). Following receptor modification (e.g., phosphorylation), the chimeric adaptor polypeptide is recruited to the receptor as shown in FIG. 8C. When in proximity to the cleavage recognition site, the cleavage moiety can cleave the recognition site to release the actuator moiety from the GMP as shown in FIG. 8D. (Page 34, col 2, para 0202). Additionally, Qi teaches “the cleavage moiety is complexed to a second adaptor polypeptide which binds the chimeric receptor polypeptide when the receptor polypeptide has undergone modification (Page 34, col 2, para 0203).
It would have been prima facie obvious to one of ordinary skill, in the art at the time of the effective filing date, to combine the teachings of a first and second fusion protein comprising an actuator moiety (i.e. anti-cancer agent which can activate or repress expression of a gene of interest) with linked cleavage site and cleavage moiety (i.e. protease), respectively, that is activated by hyperactivity of the ErbB receptor tyrosine kinase from Chung with a gene expression regulating system that uses a receptor that undergoes a modification upon binding of a ligand, thus activating/recruiting adaptor polypeptides from Qi in order to have a dual polypeptide-based gene modification system expressed in a cell that is activatable upon binding of a specific ligand rather than hyperactivity of the ErbB receptor tyrosine kinase. More specifically, by utilizing a receptor with a ligand binding domain from Qi instead of a constitutively active/hyperactive receptor from Chung, one could control the activation of the system by external factors (i.e. ligands binding to the receptor) thus allowing for the system to be used in context beyond constitutively active/hyperactive receptors. A skilled artisan would have had a reasonable expectation of success as using a dual fusion protein/polypeptide approach for gene modification that is activated by ligand binding of a cell receptor was known in the art before the effective filing date of the claimed invention to achieve elicit gene regulatory effects.
Regarding claim 2, the teachings of Chung and Qi render obvious claim 1. Moreover, Chung teaches “contacting the population of cells with an effective amount of a first fusion protein comprising a protease connected to a phosphotyrosine binding (PTB) domain (i.e. a first adaptor moiety) that selectively binds to a phosphorylated tyrosine residue on the hyperactive ErbB receptor tyrosine kinase” (page 2, para 0025), “contacting the population of cells with an effective amount of a second fusion protein comprising an SH2 domain (i.e. a second adaptor moiety) connected to a substrate comprising a cleavage site (page 2, para 0025), and “Oncogenic mutations or overexpression of ErbB (receptor) leads to its constitutive phosphorylation at cytoplasmic tyrosine residues, which then bind to phosphotyrosine-binding (PTB) and SH2 domains” (para 0238, page 23), rendering obvious the first chimeric polypeptide comprises a first adaptor moiety that is activatable to bind a first intracellular domain of the endogenous receptor and wherein the second chimeric polypeptide comprises a second adaptor moiety that is activatable to bind (i) a second intracellular domain of the endogenous receptor.
Regarding claim 4, the teachings of Chung and Qi render obvious claim 1. Moreover, Chung teaches contacting the population of cells with an effective amount of a first fusion protein comprising a protease connected to a phosphotyrosine binding (PTB) domain that selectively binds to a phosphorylated tyrosine residue (i.e. a signaling moiety) on the hyperactive ErbB receptor tyrosine kinase” (page 2, para 0025), rendering obvious wherein the first chimeric polypeptide comprises a first adaptor moiety that is activatable to bind a first downstream signaling moiety of the endogenous receptor. Chung also teaches that the two fusion proteins bind to separate signaling moieties of the receptor (Figure 7A, 7D, 7G), rendering obvious wherein the second chimeric polypeptide comprises a second adaptor moiety that is activatable to bind a second downstream signaling moiety of the endogenous receptor.
Regarding claim 14, the teachings of Chung and Qi render obvious claim 1. Moreover, Chung teaches two fusion proteins: i) a first fusion protein comprising a protease (i.e. cleavage moiety) and ii) a second fusion protein (i.e. GMP) comprising an SH2 domain connected to a substrate comprising a cleavage site recognized by the protease and an anti-cancer therapeutic agent (i.e. actuator moiety). Cleavage of the substrate by the protease of the first fusion protein releases the therapeutic agent from the second fusion protein” (page 10, para 0111), rendering obvious wherein the first chimeric polypeptide comprises the GMP, and wherein the second chimeric polypeptide comprises the cleavage moiety or wherein the second chimeric polypeptide comprises the GMP, and wherein the first chimeric polypeptide comprises the cleavage moiety.
Regarding claim 16, the teachings of Chung and Qi render obvious claim 1. Moreover, Qi teaches “the cleavage moiety is complexed with a second adaptor polypeptide that binds the chimeric receptor polypeptide in response to the receptor modification” (page 1, Summary, para 005), rendering obvious wherein the first and second chimeric polypeptides are activatable upon the contacting step (b) to form a signaling complex of the receptor.
Regarding claim 28, the teachings of Chung and Qi render obvious claim 27. Moreover, Chung teaches “contacting the population of cells with an effective amount of a first fusion protein comprising a protease connected to a phosphotyrosine binding (PTB) domain (i.e. a first adaptor moiety) that selectively binds to a phosphorylated tyrosine residue on the hyperactive ErbB receptor tyrosine kinase (i.e. first intracellular domain)” (page 2, para 0025), rendering obvious the first chimeric polypeptide comprises a first adaptor moiety that is activatable to bind a first intracellular domain of the endogenous receptor.
Regarding claim 29, the teachings of Chung and Qi render obvious claim 27 and 28. Moreover, Chung teaches “contacting the population of cells with an effective amount of a second fusion protein comprising an SH2 domain (i.e. a second adaptor moiety) connected to a substrate comprising a cleavage site (page 2, para 0025), and “Oncogenic mutations or overexpression of ErbB (receptor) leads to its constitutive phosphorylation at cytoplasmic tyrosine residues (i.e. second intracellular domain of the endogenous receptor), which then bind to phosphotyrosine-binding (PTB) and SH2 domains” (para 0238, page 23), rendering obvious wherein the second chimeric polypeptide comprises a second adaptor moiety that is activatable to bind (i) a second intracellular domain of the endogenous receptor.
Regarding claim 30, the teachings of Chung and Qi render obvious claim 27. Moreover, Chung teaches “contacting the population of cells with an effective amount of a first fusion protein comprising a protease connected to a phosphotyrosine binding (PTB) domain that selectively binds to a phosphorylated tyrosine residue (i.e. first downstream signaling moiety) on the hyperactive ErbB receptor tyrosine kinase” (page 2, para 0025), rendering obvious the first chimeric polypeptide comprises a first adaptor moiety that is activatable to bind a first downstream signaling moiety of the endogenous receptor.
Regarding claim 31, the teachings of Chung and Qi render obvious claim 27 and 30. Moreover, Chung teaches “contacting the population of cells with an effective amount of a second fusion protein comprising an SH2 domain (i.e. a second adaptor moiety) connected to a substrate comprising a cleavage site (page 2, para 0025), and “Oncogenic mutations or overexpression of ErbB (receptor) leads to its constitutive phosphorylation at cytoplasmic tyrosine residues (i.e. second intracellular domain of the endogenous receptor), which then bind to phosphotyrosine-binding (PTB) and SH2 domains” (para 0238, page 23), rendering obvious wherein the second chimeric polypeptide comprises a second adaptor moiety that is activatable to bind a (i) the first adaptor moiety, (ii) the first downstream signaling moiety, or (iii) a second downstream signaling moiety of the endogenous receptor.
Regarding claim 40, the teachings of Chung and Qi render obvious claim 27. Moreover, Chung teaches two fusion proteins: i) a first fusion protein comprising a protease (i.e. cleavage moiety) and ii) a second fusion protein (i.e. GMP) comprising an SH2 domain connected to a substrate comprising a cleavage site recognized by the protease and an anti-cancer therapeutic agent (i.e. actuator moiety). Cleavage of the substrate by the protease of the first fusion protein releases the therapeutic agent from the second fusion protein” (page 10, para 0111), rendering obvious wherein the first chimeric polypeptide comprises the GMP, and wherein the second chimeric polypeptide comprises the cleavage moiety.
Regarding claim 41, the teachings of Chung and Qi render obvious claim 27. Moreover, Chung teaches two fusion proteins: i) a first fusion protein comprising a protease (i.e. cleavage moiety) and ii) a second fusion protein (i.e. GMP) comprising an SH2 domain connected to a substrate comprising a cleavage site recognized by the protease and an anti-cancer therapeutic agent (i.e. actuator moiety). Cleavage of the substrate by the protease of the first fusion protein releases the therapeutic agent from the second fusion protein” (page 10, para 0111), rendering obvious wherein the first chimeric polypeptide comprises the GMP, and wherein the second chimeric polypeptide comprises the cleavage moiety.
Regarding claim 42, the teachings of Chung and Qi render obvious claim 27. Moreover, Qi teaches “the cleavage moiety is complexed with a second adaptor polypeptide that binds the chimeric receptor polypeptide in response to the receptor modification” (page 1, Summary, para 005), rendering obvious wherein the first and second chimeric polypeptides are activatable upon the contacting to form a signaling complex of the receptor.
Regarding claim 45, the teachings of Chung and Qi render obvious claim 27. However, Chung and Qi do not teach the receptor comprises at least a portion of T cell receptor (TCR), and wherein the TCR comprises a co-receptor of TCR, comprising CD3, CD4, or CD8.
However, Wang teaches “regulating expression of a target gene in a cell comprising (a) a transmembrane receptor comprising a ligand binding domain and a signaling domain, wherein the signaling domain activates a signaling pathway of the cell upon binding of a ligand to the ligand binding domain” (Summary, page 1, para 0005) and that the receptor can be immune receptors, such as T cell receptors (TCR) (page 50-51, para 00176) and a primary signaling domain (of the TCR) can comprise a signaling domain of CD8 (para 00280, page 99).
It would have been prima facie obvious to one of ordinary skill, in the art at the time of the effective filing date, to combine the teachings of Chung and Qi with the teaching of T cell receptors for regulating target gene expression in a cell from Wang to conditionally regulate gene expression using the dual polypeptide system via ligand-binding of a T cell receptor. One would be motivated to do so to precisely modulate gene expression through a TCR, which can be endogenous to the cell.
Regarding claim 47, the teachings of Chung and Qi render obvious claim 27. However, Chung and Qi do not teach an intracellular domain the receptor comprises at least one immunoreceptor tyrosine-based activation motif (ITAM).
Wang teaches the signaling domain comprises an immunoreceptor tyrosine-based activation motif or ITAM (para 00280, page 99).
It would have been prima facie obvious to one of ordinary skill, in the art at the time of the effective filing date, to combine the teachings of Chung and Qi with the teaching of a signaling domain comprising an ITAM from Wang to activate a signaling pathway of a cell upon binding of a ligand to the ligand binding domain. One would be motivated to do so to cause a robust signaling cascade and elicit an intracellular response that leads to gene expression modulation via the system.
Regarding claim 48, the teachings of Chung and Qi render obvious claim 27 and 28. Moreover, Qi teaches an adaptor polypeptide comprises a protein containing an SH2 domain, such as ZAP70 (page 33, para 0195), rendering obvious wherein the first adaptor moiety or the second adaptor moiety comprises one of the listed species, ZAP-70.
Regarding claim 49, the teachings of Chung and Qi render obvious claim 27.
However, Chung and Qi do not teach wherein the receptor comprises as least a portion of NKG2D.
Wang teaches a co-stimulatory signaling domain comprising a signaling domain of a molecule including NKG2D (Page 101-102, para 00283).
It would have been prima facie obvious to one of ordinary skill, in the art at the time of the effective filing date, to modify the teachings of Chung and Qi with the teachings of using a signaling domain from NKG2D in the receptor from Wang to have an alternative domain for cell activation. One would be motivated to do so to maximize approaches to activate the signaling cascade from the receptor and achieve gene expression regulation via the system.
Regarding claim 50, the teachings of Chung, Qi, and Wang render obvious claim 47. Moreover, Qi teaches an adaptor polypeptide comprises a protein containing an SH2 domain, such as ZAP70 (page 33, para 0195), rendering obvious the first adaptor moiety or the second adaptor moiety comprises DAP10, DAP12, PI3K, GRB2, VAV1, SYK, ZAP-70, a fragment thereof, or a combination thereof.
Regarding claim 52, the teachings of Chung, Qi, and Wang render obvious claim 50, 47, and 27. Moreover, Qi teaches a chimeric adaptor polypeptide comprises a
receptor-interacting serine/threonine-protein kinase 1 (RIP1 or RIPK1) (page 33, para 0196)., rendering obvious wherein the first adaptor moiety or the second adaptor moiety comprises RIPK1.
Conclusion
Claims 1, 2, 4, 14, 16, 27-31, 40-42, 45, 47-52 are rejected.
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/JULIANA IRENE CANDELARIA/Examiner, Art Unit 1634
/MARIA G LEAVITT/Supervisory Patent Examiner, Art Unit 1634