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Article: Primary T-cell-based delivery platform for in vivo synthesis of engineered proteins.

Radhakrishnan, Harikrishnan / Newmyer, Sherri L / Ssemadaali, Marvin A / Javitz, Harold S / Bhatnagar, Parijat

Bioengineering & translational medicine

2023 Volume 9, Issue 1, Page(s) e10605

Abstract: Primary T cell has been transformed into ... ...

Abstract	Primary T cell has been transformed into a
Language	English
Publishing date	2023-10-07
Publishing country	United States
Document type	Journal Article
ISSN	2380-6761
ISSN	2380-6761
DOI	10.1002/btm2.10605
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Article: Lentivirus Manufacturing Process for Primary T-Cell Biofactory Production.

Radhakrishnan, Harikrishnan / Javitz, Harold S / Bhatnagar, Parijat

Advanced biosystems

2020 Volume 4, Issue 6, Page(s) e1900288

Abstract: A process for maximizing the titer of lentivirus particles, deemed to be a necessity for transducing primary cells, is developed. Lentivirus particles, with a set of transgenes encoding an artificial cell-signaling pathway, are used to transform primary ... ...

Abstract	A process for maximizing the titer of lentivirus particles, deemed to be a necessity for transducing primary cells, is developed. Lentivirus particles, with a set of transgenes encoding an artificial cell-signaling pathway, are used to transform primary T cells as vectors for calibrated synthesis of desired proteins in situ, that is, T-cell biofactory cells. The process is also used to generate primary T cells expressing antigen-specific chimeric antigen receptors, that is, CAR T cells. The two differently engineered primary T cells are expanded and validated for their respective functions, that is, calibrated synthesis of desired proteins upon engaging the target cells, which is specific for the T-cell biofactory cells, and cytolysis of the target cells common to both types of cells. The process is compliant with current Good Manufacturing Practices and can be used to support the scale-up for clinical translation.
MeSH term(s)	Cell Engineering ; Humans ; Lentivirus ; Primary Cell Culture ; Receptors, Chimeric Antigen/genetics ; Receptors, Chimeric Antigen/metabolism ; T-Lymphocytes/cytology ; T-Lymphocytes/metabolism ; Transgenes
Chemical Substances	Receptors, Chimeric Antigen
Language	English
Publishing date	2020-05-10
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	2366-7478
ISSN	2366-7478
DOI	10.1002/adbi.201900288
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Article: Electrically regulated cell-based intervention for viral infections.

Newmyer, Sherri / Ssemadaali, Marvin A / Radhakrishnan, Harikrishnan / Javitz, Harold S / Bhatnagar, Parijat

Bioengineering & translational medicine

2022 Volume 8, Issue 2, Page(s) e10434

Abstract: This work reports on an engineered cell that-when electrically stimulated-synthesizes a desired protein, that is, ...

Abstract	This work reports on an engineered cell that-when electrically stimulated-synthesizes a desired protein, that is,
Language	English
Publishing date	2022-11-15
Publishing country	United States
Document type	Journal Article
ISSN	2380-6761
ISSN	2380-6761
DOI	10.1002/btm2.10434
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Article ; Online: Survival and Hematologic Benefits of Romiplostim After Acute Radiation Exposure Supported FDA Approval Under the Animal Rule.

Bunin, Deborah I / Javitz, Harold S / Gahagen, Janet / Bakke, James / Lane, Joan H / Andrews, Dina A / Chang, Polly Y

International journal of radiation oncology, biology, physics

2023 Volume 117, Issue 3, Page(s) 705–717

Abstract: Purpose: Patients exposed to acute high doses of ionizing radiation are susceptible to dose-dependent bone marrow depression with resultant pancytopenia. Romiplostim (RP; Nplate) is a recombinant thrombopoietin receptor agonist protein that promotes ... ...

Abstract	Purpose: Patients exposed to acute high doses of ionizing radiation are susceptible to dose-dependent bone marrow depression with resultant pancytopenia. Romiplostim (RP; Nplate) is a recombinant thrombopoietin receptor agonist protein that promotes progenitor megakaryocyte proliferation and platelet production and is an approved treatment for patients with chronic immune thrombocytopenia. The goal of our study was to evaluate the postirradiation survival and hematologic benefits of a single dose of RP with or without pegfilgrastim (PF; Neulasta, granulocyte colony stimulating factor) by conducting a well-controlled, treatment-concealed, good laboratory practice-compliant study in rhesus macaques that was compliant with the United States Food and Drug Administration Animal Rule regulatory approval pathway. Methods and materials: Irradiated male and female rhesus macaques (20/sex in each of 3 groups: control, RP, and RP + PF) were subcutaneously administered vehicle or RP (5 mg/kg, 10 mL/kg) on day 1 in the presence or absence of 2 doses of PF (0.3 mg/kg, 0.03 mL/kg, days 1 and 8). Total body radiation (680 cGy, 50 cGy/min from cobalt-60 gamma ray source) occurred 24 ± 2 hours previously at a dose targeting 70% lethality for the control cohort over 60 days. The study examined 60-day survival postirradiation as the primary endpoint. Secondary endpoints included incidence, severity, and duration of thrombocytopenia and neutropenia, other hematology parameters, coagulation parameters, and body weight change to provide insights into potential mechanisms of action. Results: Compared with sham-treated controls, treated animals demonstrated a 40% to 55% survival benefit compared with controls, less severe clinical signs, reduced incidence of thrombocytopenia and/or neutropenia, earlier hematologic recovery, and reduced morbidity from bacterial infection. Conclusions: These results were pivotal in obtaining Food and Drug Administration approval in January 2021 for RP's new indication as a single administration therapy to increase survival in adults and pediatric patients acutely exposed to myelosuppressive doses of radiation.
MeSH term(s)	Adult ; Animals ; Humans ; Male ; Female ; Child ; Macaca mulatta ; Recombinant Proteins ; Thrombocytopenia/drug therapy ; Thrombocytopenia/etiology ; Neutropenia/drug therapy ; Hematology ; Radiation Exposure
Chemical Substances	romiplostim (GN5XU2DXKV) ; Recombinant Proteins
Language	English
Publishing date	2023-05-22
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	197614-x
ISSN	1879-355X ; 0360-3016
ISSN (online)	1879-355X
ISSN	0360-3016
DOI	10.1016/j.ijrobp.2023.05.008
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Article ; Online: The progression of radiation injury in a Wistar rat model of partial body irradiation with ∼5% bone marrow shielding.

Beach, Tyler / Bakke, James / Riccio, Ed / Javitz, Harold S / Nishita, Denise / Kapur, Shweta / Bunin, Deborah I / Chang, Polly Y

International journal of radiation biology

2023 Volume 99, Issue 7, Page(s) 1080–1095

Abstract: Purpose: To describe the dose response relationship and natural history of radiation injury in the Wistar rat and its suitability for use in medical countermeasures (MCM) testing.: Materials & methods: In two separate studies, male and female rats ... ...

Abstract	Purpose: To describe the dose response relationship and natural history of radiation injury in the Wistar rat and its suitability for use in medical countermeasures (MCM) testing. Materials & methods: In two separate studies, male and female rats were exposed to partial body irradiation (PBI) with 5% bone marrow sparing. Animals were X-ray irradiated from 7 to 12 Gy at 7-10 weeks of age. Acute radiation syndrome (ARS) survival at 30 days and delayed effects of acute radiation exposure (DEARE) survival at 182 days were assessed. Radiation effects were determined by clinical observations, body weights, hematology, clinical chemistry, magnetic resonance imaging of lung, whole-body plethysmography, and histopathology. Results: Rats developed canonical ARS responses of hematopoietic atrophy and gastrointestinal injury resulting in mortality at doses ≥8Gy in males and ≥8.5 Gy in females. DEARE mortality occurred at doses ≥8Gy for both sexes. Findings indicate lung, kidney, and/or liver injury, and persistent hematological dysregulation, revealing multi-organ injury as a DEARE. Conclusion: The Wistar rat PBI model is suitable for testing MCMs against hematopoietic and gastrointestinal ARS. DEARE multi-organ injury occurred in both sexes irradiated with 8-9Gy, also suggesting suitability for polypharmacy studies addressing the combination of ARS and DEARE injury.
MeSH term(s)	Male ; Female ; Rats ; Animals ; Bone Marrow/radiation effects ; Rats, Wistar ; Acute Radiation Syndrome/etiology ; Acute Radiation Syndrome/pathology ; Gastrointestinal Tract/radiation effects ; Hematopoietic System
Language	English
Publishing date	2023-03-24
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	3065-x
ISSN	1362-3095 ; 0020-7616 ; 0955-3002
ISSN (online)	1362-3095
ISSN	0020-7616 ; 0955-3002
DOI	10.1080/09553002.2023.2188937
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Article: Genetically engineered pair of cells for serological testing and its application for SARS-CoV-2.

Ssemadaali, Marvin A / Arredondo, Juan / Buser, Elise A / Newmyer, Sherri / Radhakrishnan, Harikrishnan / Javitz, Harold S / Dandekar, Satya / Bhatnagar, Parijat

Bioengineering & translational medicine

2023 Volume 8, Issue 3, Page(s) e10508

Abstract: We have developed a serology test platform for identifying individuals with prior exposure to specific viral infections and provide data to help reduce public health risks. The serology test composed of a pair of cell lines engineered to express either a ...

Abstract	We have developed a serology test platform for identifying individuals with prior exposure to specific viral infections and provide data to help reduce public health risks. The serology test composed of a pair of cell lines engineered to express either a viral envelop protein (Target Cell) or a receptor to recognize the Fc region of an antibody (Reporter Cell), that is,
Language	English
Publishing date	2023-03-24
Publishing country	United States
Document type	Journal Article
ISSN	2380-6761
ISSN	2380-6761
DOI	10.1002/btm2.10508
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Article ; Online: Delayed effects of radiation exposure in a C57L/J mouse model of partial body irradiation with ~2.5% bone marrow shielding.

Beach, Tyler / Bakke, James / McDonald, J Tyson / Riccio, Edward / Javitz, Harold S / Nishita, Denise / Kapur, Shweta / Bunin, Deborah I / Chang, Polly Y

Frontiers in public health

2024 Volume 12, Page(s) 1349552

Abstract: Introduction: Mouse models of radiation injury are critical to the development of medical countermeasures (MCMs) against radiation. Now that MCMs against hematopoietic acute radiation syndrome (H-ARS) have achieved regulatory approval, attention is ... ...

Abstract	Introduction: Mouse models of radiation injury are critical to the development of medical countermeasures (MCMs) against radiation. Now that MCMs against hematopoietic acute radiation syndrome (H-ARS) have achieved regulatory approval, attention is shifting to develop MCMs against the adverse effects of gastrointestinal acute radiation syndrome (GI-ARS) and delayed effects of acute radiation exposure (DEARE). The C57L/J mouse model of partial body irradiation (PBI) with 2.5% bone marrow shielding (BM2.5) is being leveraged to examine both GI-ARS and DEARE effects. Within days of PBI, mice may develop H- and GI-ARS followed several months later by DEARE as a multi-organ injury, which typically involves the lung and kidney (L- and K-DEARE, respectively). The objective of this manuscript is to describe the dose response relationship and progression of radiation injury in the C57L/J mouse and to evaluate its suitability for use in DEARE MCM testing. Materials and methods: In two separate studies conducted over 2 years, male and female C57L/J mice were exposed to PBI BM2.5 with one hindlimb shielded from radiation, representing ~2.5% bone marrow shielding/sparing. Mice were X-ray irradiated at doses ranging from 9 to 13 Gy at 10 to 12 weeks of age for the purposes of assessing ARS survival at 30 days and DEARE survival at 182 days post-irradiation. Clinical indicators of ARS and DEARE were determined by clinical observations, body weights, hematology, clinical chemistry, magnetic resonance imaging (MRI) of lung, and histopathology of selected tissues. Results: C57L/J mice developed canonical ARS responses of hematopoietic atrophy and gastrointestinal injury resulting in dose dependent mortality at doses ≥11 Gy between 1- and 15-days post-irradiation. In animals that survived ARS, DEARE associated mortality occurred in dose dependent fashion at ≥9 Gy for both sexes between 60- and 159-days post-irradiation with histopathology examinations indicating lung injury as the primary cause of death in moribund animals. Conclusion: The PBI BM2.5 C57L/J mouse model reliably produced known H- and GI-ARS effects at doses greater than those resulting in DEARE effects. Because of this, the C57L/J mouse can be used to test MCMs against L-DEARE injury, while avoiding ARS associated mortality.
MeSH term(s)	Male ; Female ; Mice ; Animals ; Bone Marrow/pathology ; Bone Marrow/radiation effects ; Acute Radiation Syndrome/etiology ; Acute Radiation Syndrome/pathology ; Disease Models, Animal ; Lung/pathology
Language	English
Publishing date	2024-03-12
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2711781-9
ISSN	2296-2565 ; 2296-2565
ISSN (online)	2296-2565
ISSN	2296-2565
DOI	10.3389/fpubh.2024.1349552
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Article ; Online: NK-Cell Biofactory as an Off-the-Shelf Cell-based Vector for Targeted In Situ Synthesis of Engineered Proteins.

Repellin, Claire E / Ssemadaali, Marvin A / Newmyer, Sherri / Radhakrishnan, Harikrishnan / Javitz, Harold S / Bhatnagar, Parijat

Advanced biology

2021 Volume 5, Issue 7, Page(s) e2000298

Abstract: The NK-92MI, a fast-growing cytolytic cell line with a track record of exerting clinical efficacy, is transformed into a vector for synthesizing calibrated amounts of desired engineered proteins at our disease site, that is, NK-cell Biofactory. This ... ...

Abstract	The NK-92MI, a fast-growing cytolytic cell line with a track record of exerting clinical efficacy, is transformed into a vector for synthesizing calibrated amounts of desired engineered proteins at our disease site, that is, NK-cell Biofactory. This provides an allogeneic option to the previously published T-cell-based living vector that is limited by high manufacturing cost and product variability. The modularity of this pathway, which combines a "target" receptor with an "effector" function, enables reprogramming of the NK-cell Biofactory to target diseases with specific molecular biomarkers, such as cancer, viral infections, or auto-immune disorders, and overcome barriers that may affect the advancement of NK-cell therapies.
MeSH term(s)	Humans ; Immunotherapy, Adoptive ; Killer Cells, Natural ; Neoplasms ; Receptors, Chimeric Antigen ; T-Lymphocytes
Chemical Substances	Receptors, Chimeric Antigen
Language	English
Publishing date	2021-04-19
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ISSN	2701-0198
ISSN (online)	2701-0198
DOI	10.1002/adbi.202000298
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Article ; Online: Cell-Based Platform for Antigen Testing and Its Application for SARS-CoV-2 Infection.

Ssemadaali, Marvin A / Newmyer, Sherri / Radhakrishnan, Harikrishnan / Arredondo, Juan / Javitz, Harold S / Dandekar, Satya / Bhatnagar, Parijat

Microbiology spectrum

2022 Volume 10, Issue 4, Page(s) e0073122

Abstract: We have engineered a cell that can be used for diagnosing active severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections. Isolation of individuals with active infections offers an effective solution for mitigating pandemics. However, the ... ...

Abstract	We have engineered a cell that can be used for diagnosing active severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections. Isolation of individuals with active infections offers an effective solution for mitigating pandemics. However, the implementation of this practice requires robust infrastructure for rapid and intuitive testing, which is currently missing in our communities. To address this need, we engineered a fast-growing cell line into a cell-based antigen test platform for emerging viruses, i.e., DxCell, that can be rapidly deployed in decentralized health care facilities for continuous testing. The technology was characterized using cells engineered to present spike glycoprotein of SARS-CoV-2 (SARS-CoV-2-Sgp-cells) and Calu-3 host cells infected with competent SARS-CoV-2. Preclinical validation was conducted by directly incubating the DxCell with oropharyngeal swabs from mice infected with SARS-CoV-2. No sample preparation steps are necessary. The DxCell quantitatively detected the SARS-CoV-2-Sgp-cells within 1 h (
MeSH term(s)	Animals ; COVID-19/diagnosis ; COVID-19 Testing ; Mice ; Pandemics ; SARS-CoV-2/genetics ; Specimen Handling
Language	English
Publishing date	2022-07-19
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2807133-5
ISSN	2165-0497 ; 2165-0497
ISSN (online)	2165-0497
ISSN	2165-0497
DOI	10.1128/spectrum.00731-22
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Article ; Online: Romiplostim (Nplate

Bunin, Deborah I / Bakke, James / Green, Carol E / Javitz, Harold S / Fielden, Mark / Chang, Polly Y

International journal of radiation biology

2019 Volume 96, Issue 1, Page(s) 145–154

Abstract: Purpose: ...

Abstract	Purpose:
MeSH term(s)	Animals ; Blood Platelets/cytology ; Blood Platelets/drug effects ; Blood Platelets/radiation effects ; Dose-Response Relationship, Drug ; Drug Interactions ; Female ; Filgrastim/pharmacology ; Male ; Medical Countermeasures ; Mice ; Mice, Inbred C57BL ; Platelet Count ; Polyethylene Glycols/pharmacology ; Receptors, Fc ; Recombinant Fusion Proteins/pharmacokinetics ; Recombinant Fusion Proteins/pharmacology ; Survival Analysis ; Thrombopoietin/pharmacokinetics ; Thrombopoietin/pharmacology ; X-Rays/adverse effects
Chemical Substances	Receptors, Fc ; Recombinant Fusion Proteins ; pegfilgrastim (3A58010674) ; Polyethylene Glycols (3WJQ0SDW1A) ; Thrombopoietin (9014-42-0) ; romiplostim (GN5XU2DXKV) ; Filgrastim (PVI5M0M1GW)
Language	English
Publishing date	2019-04-29
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	3065-x
ISSN	1362-3095 ; 0020-7616 ; 0955-3002
ISSN (online)	1362-3095
ISSN	0020-7616 ; 0955-3002
DOI	10.1080/09553002.2019.1605465
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