Browsing by Author "Takundwa, Mutsa M"

Now showing 1 - 11 of 11
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    Advances in CRISPR-Cas systems for blood cancer
    (2024-08) Monchusi, Bernice A; Dube, Phumzile; Takundwa, Mutsa M; Kenmogne, Vanelle L; Thimiri Govinda Raja, Deepak B
    CRISPR-Cas systems have revolutionised precision medicine by enabling personalised treatments tailored to an individual's genetic profile. Various CRISPR technologies have been developed to target specific disease-causing genes in blood cancers, and some have advanced to clinical trials. Although some studies have explored the in vivo applications of CRISPR-Cas systems, several challenges continue to impede their widespread use. Furthermore, CRISPR-Cas technology has shown promise in improving the response of immunotherapies to blood cancers. The emergence of CAR-T cell therapy has shown considerable success in the targeting and correcting of disease-causing genes in blood cancers. Despite the promising potential of CRISPR-Cas in the treatment of blood cancers, issues related to safety, ethics, and regulatory approval remain significant hurdles. This comprehensive review highlights the transformative potential of CRISPR-Cas technology to revolutionise blood cancer therapy.
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    Application of drug repurposing-based precision medicine platform for leukaemia patient treatment
    (2022-10) Kenmogne, Vanelle L; Nweke, EE; Takundwa, Mutsa M; Fru, PN; Thimiri Govindaraj, Deepak B
    Drug resistance in leukaemia is a major problem that needs to be addressed. Precision medicine provides an avenue to reduce drug resistance through a personalised treatment plan. It has helped to better stratify patients based on their molecular profile and therefore improved the sensitivity of patients to a given therapeutic regimen. However, therapeutic options are still limited for patients who have already been subjected to many lines of chemotherapy. The process of designing and developing new drugs requires significant resources, including money and time. Drug repurposing has been explored as an alternative to identify effective drug(s) that could be used to target leukaemia and lessen the burden of drug resistance. The drug repurposing process usually includes preclinical studies with drug screening and clinical trials before approval. Although most of the repurposed drugs that have been identified are generally safe for leukaemia treatment, they seem not to be good candidates for monotherapy but could have value in combination with other drugs, especially for patients who have exhausted therapeutic options. In this review, we highlight precision medicine in leukaemia and the role of drug repurposing. Specifically, we discuss the several screening methods via chemoinformatic, in vitro, and ex vivo that have facilitated and accelerated the drug repurposing process.
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    DirectedCHO: A new miniaturized directed evolution process for phenotype stability trial test of CHO cells before bioreactor scale-up
    (2024-08) Thimiri Govindaraj, Deepak B; Musasira, Nyasha; Takundwa, Mutsa M
    Most of the biopharmaceuticals that are currently on the market are expressed using the Chinese Hamster Ovary (CHO) cell lines. However, the production yield of these biopharmaceuticals is affected due to CHO cellular heterogeneity and challenges in adaptability during the bioreactor scale-up stage. In this communication, we report the protocol for the miniaturized directed evolution process for CHO cells. The results of the directed evolution process would guide adapting the CHO cell line before bioreactor scale-up. With our approach, we have established the protocol that can be used to streamline superior CHO cell lines for biopharmaceutical production which would be the first of its kind in Africa. Our directed evolution protocol includes a method for a low-cost multiplex directed evolution process that can be used on CHO cells using 20 stressors in 8 concentrations and provides stable trial results for the scale-up process. Using our process, we can provide a simple consumable kit that manufacturers can use for the CHO cell phenotype stability test before the scale-up process. With our approach, we would further develop a platform that can streamline superior CHO cell lines for biopharmaceutical production. This approach would be the first of its kind in South Africa/ Africa.
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    Establishment of ex vivo drug sensitivity screening platform for leukaemia and multiple myeloma using a South African patient cohort
    (2023-10) Kenmogne, VL; Takalani, AMT; Nweke, EE; Takundwa, Mutsa M; Fabian, June; Maher, Heather; Du Toit, Justin; Philip-Cherian, V; Fru, PF; Govindaraj, Deepak BT
    Our objective is to develop a functional precision medicine platform designed to directly identify tailored drug regimens that target individual patient cancer cells and give benefit to the same donors by supporting clinical decision-making. We demonstrate our ex vivo drug sensitivity screening platform for precision medicine using Leukaemia and Multiple Myeloma samples from a South African patient cohort as proof of concept.
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    Establishment of ex vivo drug sensitivity screening platform for leukaemia and multiple myeloma using a South African patient cohort
    (2023-10) Kenmogne, VL; Takalani, AMT; Nweke, EE; Takundwa, Mutsa M; Fabian, June; Maher, Heather; Du Toit, Justin; Philip-Cherian, V; Fru, PF; Govindaraj, Deepak BT
    Our objective is to develop a functional precision medicine platform designed to directly identify tailored drug regimens that target individual patient cancer cells and give benefit to the same donors by supporting clinical decision-making. We demonstrate our ex vivo drug sensitivity screening platform for precision medicine using Leukaemia and Multiple Myeloma samples from a South African patient cohort as proof of concept.
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    Evaluation of ssDNA aptamer binding to Aurora Kinase A and its cytotoxic effects on HeLa and MCF-7 cancer cell lines
    (2025-07) Woldekidan, Haregewoin B; Takundwa, Mutsa M; Thimiri Govinda Raj, Deepak B; Woldesemayat, AA
    Purpose: To investigate the binding potential of ssDNA aptamer to Aurora kinase A (AURKA) and assess its inhibitory effect on HeLa and MCF-7 cell lines. Methods: A detailed analysis was conducted on the binding between aptamer and AURKA protein using HDOCK and HADDOCK, with 2D interaction obtained by LIGPLOT+ v1.4. Binding of FAMlabelled ssDNA aptamer to AURKA was analyzed by EVOS-5000 imaging and the inhibitory effects of aptamer and aptamer-drug complexes (Bleomycin Sulfate, Metformin, and Mitomycin C) at different concentrations and times were evaluated by PrestoBlue assay. Results: Docking results showed a HADDOCK score of -82.0 ± 6.4 and a Z-score of -1.5. The HDOCK result showed best docking result (-327.13) at a confidence interval of 0.9719. Free aptamer and its drug complexes effectively inhibited HeLa cell growth, with the highest cytotoxic effect observed at higher concentrations. Free aptamer achieved the highest drug sensitivity score (DSS: 40.7) in HeLa, followed by its bleomycin sulfate and metformin complex (DSS: 31.6). However, MCF-7 exhibited lower sensitivity and DSS score. Conclusion: ssDNA aptamer shows potential in cancer detection and therapy. In vitro and in vivo studies are needed to ascertain the potential of ssDNA aptamers in cancer therapy.
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    Novel strategies for drug repurposing
    (2024) Takundwa, Mutsa M; Thimiri Govindaraj, Deepak B
    Synthetic biology, precision medicine, and nanobiotechnology are the three main emerging areas that drive translational innovation toward commercialization. There are several strategies used in precision medicine and drug repurposing is one of the key approaches as it addresses the challenges in drug discovery (high cost and time). Here, we provide a perspective on various new approaches to drug repurposing for cancer precision medicine. We report here our optimized wound healing methodology that can be used to validate drug sensitivity and drug repurposing. Using HeLa as our benchmark, we demonstrated that the assay can be applied to identify drugs that limit cell proliferation. From a future perspective, this assay can be expanded to ex vivo culturing of solid tumors in 2D culture and leukemia in 3D culture.
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    Patents, ethics, biosafety and regulation using CRISPR technology
    (2021-02) Nxumalo, Precious Z; Takundwa, Mutsa M; Thimiri Govindaraj, Deepak B
    In this review chapter, we provide full comprehensive analysis on the patent, ethics and biosafety regulation with respect to the application of CRISPR technology in mammalian systems. We focused on recent development in CRISPR technology and its patent landscape between countries such as US, European Union, China and Australia. Further, we emphasized on the current scenarios on the ethics regulations with respect to CRISPR research, its applicability in patent and technology transfer. Finally, we elaborated on the biosafety regulation on CRISPR/Cas9 technology application in both mammalian and non-mammalian host system.
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    Recent advances in genome annotation and synthetic biology for the development of microbial chassis
    (2023-12) Hamese, Saltiel; Mugwanda, Kanganwiro; Takundwa, Mutsa M; Prinsloo, E; Thimiri Govindaraj, Deepak B
    This article provides an overview of microbial host selection, synthetic biology, genome annotation, metabolic modeling, and computational methods for predicting gene essentiality for developing a microbial chassis. This article focuses on lactic acid bacteria (LAB) as a microbial chassis and strategies for genome annotation of the LAB genome. As a case study, Lactococcus lactis is chosen based on its well-established therapeutic applications such as probiotics and oral vaccine development. In this article, we have delineated the strategies for genome annotations of lactic acid bacteria. These strategies also provide insights into streamlining genome reduction without compromising the functionality of the chassis and the potential for minimal genome chassis development. These insights underscore the potential for the development of efficient and sustainable synthetic biology systems using streamlined microbial chassis with minimal genomes.
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    The first-in-Africa ex vivo drug sensitivity testing platform identifies novel drug combinations for South African leukaemia patient cohort
    (2025-03) Kenmogne, VL; Takundwa, Mutsa M; Nweke, EE; Monchusi, Bernice A; Dube, Phumzile; Maher, H; Du Toit, J; Philip-Cherian, V; Fru, PN; Thimiri Govinda Raj, Deepak B
    In South Africa, leukemia remains a major health concern, posing significant challenges in treatment due to its varied subtypes. There is an unmet need for a testing pipeline that can identify drug effects on patient samples in an ex-vivo setting. Using the pilot study with South African patient samples, this paper reports the development of a drug-sensitivity testing pipeline for studying the drug effects in leukemia patient-derived cells. Forty-one (41) patients with Acute myeloid leukemia (AML) (n = 7), Chronic myelogenous leukemia (CML) (n = 30), and Chronic lymphocytic leukemia (CLL) (n = 4) were recruited for this study. Thirty (30) FDA-approved drugs were utilized for single drug sensitivity screening (DSS) on leukemia patient-derived cells with drug concentrations (1–1000 nM). The single DSS showed a distinct sensitivity pattern with different profiles among patients of the same subtype, confirming the need for precision therapy. This study observed irinotecan, used in solid tumour treatment, demonstrated efficacy in PBMCs in many patient samples compared to conventional leukemia drugs such as nilotinib. For drug combination studies, ten clinically relevant drugs were selected and tested based on the results of single drug sensitivity tests. This pilot study marks a crucial stride towards revolutionizing leukemia treatment in South Africa through an innovative ex vivo drug sensitivity testing platform. This pioneering initiative forms the basis for tailored and effective treatment options holding promise for more personalized treatment. Further exploration and validation of these findings could significantly contribute to cancer precision medicine efforts in South Africa.
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    Treatment strategies for multiple myeloma treatment and the role of high-throughput screening for precision cancer therapy
    (2023-05) Malise, TTA; Nweke, EE; Takundwa, Mutsa M; Fru, PF; Thimiri Govindaraj, Deepak B
    In the past few years, development of approved drug candidates has improved the disease management of multiple myeloma (MM). However, due to drug resistance, some of the patients do not respond positively, while some of the patients acquire drug resistance, thereby these patients eventually relapse. Hence, there are no other therapeutic options for multiple myeloma patients. Therefore, this necessitates a precision-based approach to multiple myeloma therapy. The use of patient's samples to test drug sensitivity to increase efficacy and reduce treatment-related toxicities is the goal of functional precision medicine. Platforms such as high-throughput-based drug repurposing technology can be used to select effective single drug and drug combinations based on the efficacy and toxicity studies within a time frame of couple of weeks. In this article, we describe the clinical and cytogenetic features of MM. We highlight the various treatment strategies and elaborate on the role of high-throughput screening platforms in a precision-based approach towards clinical treatment.