Genetic Engineering Publications - GEG Tech top picks
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How to supercharge cancer-fighting cells: give them stem-cell skills - Nature

How to supercharge cancer-fighting cells: give them stem-cell skills - Nature | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
The bioengineered immune players called CAR T cells last longer and work better if pumped up with a large dose of a protein that makes them resemble stem cells.
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Keeping cells active long enough to eliminate cancer has proved difficult, particularly in solid tumors such as those of the breast and lung. Scientists are therefore looking for better ways to help CAR T cells multiply faster and last longer in the body. To this end, researchers compared samples of CAR T cells used to treat people with leukemia. They analyzed the role of cellular proteins that regulate gene activity and serve as master switches in T cells. They discovered a set of 41 genes that were more active in CAR T cells associated with a good response to treatment than in cells associated with a poor response. All 41 genes appeared to be regulated by a master protein called FOXO1. The researchers then engineered the CAR T cells to produce more FOXO1 than usual. Gene activity in these cells began to resemble that of memory T stem cells, which recognize cancer and respond quickly to it. The researchers then injected the modified cells into mice with different types of cancer. Extra FOXO1 enabled the CAR T cells to better reduce both solid tumors and blood cancers. Moreover, another team also reached the same conclusion by working on gene activity analysis in CAR T cells and also discovered that IL-15 activated genes associated with FOXO1. 

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Genetic Engineering Publications - GEG Tech top picks
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Trisomic rescue via allele-specific multiple chromosome cleavage using CRISPR-Cas9 in trisomy 21 cells | PNAS Nexus | Oxford Academic

Trisomic rescue via allele-specific multiple chromosome cleavage using CRISPR-Cas9 in trisomy 21 cells | PNAS Nexus | Oxford Academic | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Abstract. Human trisomy 21, responsible for Down syndrome, is the most prevalent genetic cause of cognitive impairment and remains a key focus for prenatal
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The present study demonstrates that allele-specific multiple chromosome cleavage by clustered regularly interspaced palindromic repeats Cas9 can achieve trisomy rescue by eliminating the target chromosome from human trisomy 21 induced pluripotent stem cells and fibroblasts.

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Advancing the genetic engineering toolbox by combining AsCas12a knock-in mice with ultra-compact screening | Nature Communications

Advancing the genetic engineering toolbox by combining AsCas12a knock-in mice with ultra-compact screening | Nature Communications | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Cas12a is a next-generation gene editing tool that enables multiplexed gene targeting. Here, we present a mouse model that constitutively expresses enhanced Acidaminococcus sp. Cas12a (enAsCas12a) linked to an mCherry fluorescent reporter. We demonstrate efficient single and multiplexed gene editing in vitro, using primary and transformed cells from enAsCas12a mice. We further demonstrate successful in vivo gene editing, using normal and cancer-prone enAsCas12a stem cells to reconstitute the haematopoietic system of wild-type mice. We also present compact, genome-wide Cas12a knockout libraries, with four crRNAs per gene encoded across one (Scherzo) or two (Menuetto) vectors, and demonstrate the utility of these libraries across methodologies: in vitro enrichment and drop-out screening in lymphoma cells and immortalised fibroblasts, respectively, and in vivo screens to identify lymphoma-driving events. Finally, we demonstrate CRISPR multiplexing via simultaneous gene knockout (via Cas12a) and activation (via dCas9-SAM) using primary T cells and fibroblasts. Our enAsCas12a mouse and accompanying crRNA libraries enhance genome engineering capabilities and complement current CRISPR technologies. Cas12a represents the next generation of gene editing. Here, the authors present the generation and validation of a Cas12a transgenic mouse model. Additionally, the authors create whole-genome Cas12a knockout libraries, and demonstrate their utility across multiple in vitro and in vivo screens.
BigField GEG Tech's insight:

Cas enzymes are needed to cut specific sections of DNA or RNA in experiments using the CRISPR tool. CRISPR is a revolutionary gene-editing tool widely used in cancer research and is currently in the early stages of clinical application in patients. Over the past decade, the Cas enzyme most commonly used with CRISPR, Cas9, has led to many important discoveries in medical research. Today, cancer researchers have developed a next-generation gene-editing tool for modelling and interrogating human disease. These researchers have generated a new preclinical model expressing an improved version of a new genome-engineering enzyme called Cas12a. They also identified the genes that led to accelerated lymphoma growth in the preclinical model using unique CRISPR ‘libraries’ of Cas12a-compatible whole mouse genomes.

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New findings could make off-the-shelf CAR T cell therapy a reality

New findings could make off-the-shelf CAR T cell therapy a reality | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
CAR T cell therapy is one of the most promising new cancer treatments to emerge in recent years. It involves removing a patient's own immune T cells and engineering them to recognize specific targets on the surface of the cancer cell.
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One of the main limitations of CAR-T cells is that the cells are taken from the patient and have to be customized for treatment. This forces patients to wait for their cells to be modified for infusion, which is precious time they may not have. Research has demonstrated a new advance that could allow standard CAR T cells supplied by healthy donors to be used and stored to be ready as soon as the patient needs them. The research has identified a way of modifying donor CAR T cells, known as allogeneic CAR T cells, so that they are accepted by the patient who receives them and persists to fight cancer. The new approach involves equipping CAR T cells with a protein called Nef. This protein has been identified in HIV and is used to evade detection by the immune system. It acts in two ways: it reduces a protein called HLA-I on the surface of CAR-T cells and helps prevent cell suicide called apoptosis in CAR-T cells. The researchers showed that inserting Nef into donor CAR-T cells enabled the cells to survive and remain effective in a mouse cancer model. 

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EZH2 inhibitors enhance effectiveness of cancer immunotherapies

EZH2 inhibitors enhance effectiveness of cancer immunotherapies | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
An emerging class of anticancer drugs called EZH2 inhibitors may greatly enhance the potency of some cancer immunotherapies, according to a preclinical study led by Weill Cornell Medicine lymphoma researchers.
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T-cell immunotherapies, including modified anti-cancer T cells called CAR-T cells, have been shown to reduce or eliminate signs of cancer in some patients with lymphomas and other malignant blood tumors. In most cases, however, signs of cancer eventually reappear. A study published in Cancer Cell suggests that inhibiting EZH2 could help improve the potency and durability of these immunotherapies. EZH2 is an enzyme that normally helps program cellular behavior by controlling the expression of specific genes. Mutations in the EZH2 gene, which make the enzyme more active, are now recognized as common features of lymphoma, and inhibition of this enzyme has been shown to benefit lymphoma patients even when they have non-mutant EZH2. In the study, the researchers found that EZH2 inhibition enhanced the effects of immunotherapies not only by making lymphoma cells more visible to them, but also through multiple other mechanisms, including reducing immunosuppressive regulatory T cells and reprogramming anti-cancer T cells in a way that makes their activity more durable.

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New technology guides immune cells to fight brain cancer and multiple sclerosis

New technology guides immune cells to fight brain cancer and multiple sclerosis | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Researchers have found a way to program immune cells to attack glioblastoma and treat the inflammation of multiple sclerosis in mice.
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Brain cancers are among the most difficult to treat. Surgery and chemotherapy are risky, and drugs often fail to penetrate the brain - a major limitation of CAR-T cancer therapies. To circumvent these problems, scientists have developed a “molecular GPS” for immune cells that guides them with a first molecular code to the brain and a second code to the tumor. The first molecular code is a protein called brevican, which helps form the gelatinous structure of the brain and only appears there. For the second code, they used two proteins found in most brain cancers. The scientists programmed the immune cells to attack only if they first detected brevican, and then either of the brain cancer proteins. They showed that immune cells could eliminate a deadly brain tumor called glioblastoma and prevent recurrence. They have also used these cells to reduce inflammation in a mouse model of multiple sclerosis. This technology will soon be tested in a clinical trial on people with glioblastoma.

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December 9, 2024 7:03 AM
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New approach reinvigorates exhausted T cells to improve tumor control

New approach reinvigorates exhausted T cells to improve tumor control | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
As cancer cells grow, they pump out metabolic byproducts such as lactic acid into the tumor microenvironment. Exhausted T cells -; which have lost their cancer-fighting oomph -; consume this lactic acid, which further saps their energy, according to new research from the University of Pittsburgh and UPMC Hillman Cancer Center.
BigField GEG Tech's insight:

With continued exposure to tumours, T lymphocytes progressively lose their effectiveness due to the expression of co-inhibitory receptors that act as brakes. Exhausted T lymphocytes, which still retain some anti-cancer function, can deteriorate further until they reach a state of terminal exhaustion. Researchers found that a solute transporter called MCT11, which imports lactic acid, was significantly increased in terminally exhausted T cells compared with their progenitor versions, suggesting that lactic acid contributes to loss of function. When the researchers deleted the gene encoding MCT11 in mice or blocked the protein with a monoclonal antibody, the T cells ingested less lactic acid. They showed improved functionality and tumour control in mouse models of melanoma, colorectal carcinoma and head and neck cancer. The researchers found that the MCT11 antibody promoted tumour killing in mice when administered alone but was even more effective when combined with anti-PD1. Through their new spin-out company, Delgoffe and Peralta are now working to optimise the MCT antibody for efficacy on human T cells to test it in future clinical trials. 

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November 18, 2024 6:41 AM
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New platform mimics immune interactions to boost cancer treatment

New platform mimics immune interactions to boost cancer treatment | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Imagine a world where your own immune cells are transformed into cancer-fighting superheroes.
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Current methods of activating T cells do not sufficiently mimic the natural environment in which they interact with another key population of immune cells, a crucial connection for activating T cells and enhancing their ability to fight cancer. In a recent study published in Nature Nanotechnology, researchers introduced a powerful tool to overcome this limitation. Their innovative platform combines a flexible material called graphene oxide with antibodies to closely mimic the natural interactions between immune cells. The researchers anchored two specific antibodies to the graphene oxide. In just 12 days, their platform increased the expansion of T lymphocytes in a blood cell culture by a staggering 100-fold. The technology also enhanced the efficiency of immune cell engineering, resulting in a five-fold increase in CAR-T cell production compared to the standard process. The team also identified several biochemical pathways crucial to T-cell signaling and function activated by their technology, leading to increased growth and efficiency. This breakthrough has the potential to significantly improve CAR-T cell therapy, making it more effective and accessible while also advancing other emerging treatments.

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October 30, 2024 7:29 AM
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Buoyed by ‘milestone’ clinical result, RNA editing is poised to treat diseases | Science | AAAS

Buoyed by ‘milestone’ clinical result, RNA editing is poised to treat diseases | Science | AAAS | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Multiple approaches aim to counteract mutant genes by fixing the faulty RNA they produce
BigField GEG Tech's insight:

The field of RNA editing may be in its infancy, but pharma companies are already testing its use in some types of eye disease and cancer.

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Innovative CAR-T cells effectively target glioblastoma with reduced toxicity

Innovative CAR-T cells effectively target glioblastoma with reduced toxicity | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Glioblastoma is the most common and most aggressive primary brain tumor, with an average survival after diagnosis of less than two years, and against which current treatments remain ineffective.
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Glioblastoma is the most common and aggressive primary brain tumor, with an average survival after diagnosis of less than two years, and against which current treatments remain ineffective. In recent years, immunotherapies have given patients hope, albeit with relatively modest success. One research team has succeeded in identifying a specific marker, PTPRZ1, on the surface of tumour cells, and generating CAR-T cells to destroy them. Moreover, this CAR-T cell-based therapy appears to be able to target diseased tumor cells that do not carry this specific marker, while sparing healthy cells. The team tested the treatment in vivo on mouse models of human glioblastoma. Tumor growth was controlled, remarkably prolonging the life of the mice without signs of toxicity. These results, published in the journal Cancer Immunology Research, represent a first step towards the development of clinical trials with human patients. 

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October 4, 2024 5:22 AM
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CRISPR helps brain stem cells regain youth in mice

CRISPR helps brain stem cells regain youth in mice | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Disabling a gene involved in metabolism rejuvenates cells’ ability to spin off new neurons.
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Clues to keeping the brain’s regenerative cells youthful and energetic into old age have emerged by applying CRISPR gene editing to mice

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September 23, 2024 8:16 AM
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Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells | Nature Medicine

Targeting axonal guidance dependencies in glioblastoma with ROBO1 CAR T cells | Nature Medicine | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Resistance to genotoxic therapies and tumor recurrence are hallmarks of glioblastoma (GBM), an aggressive brain tumor. In this study, we investigated functional drivers of post-treatment recurrent GBM through integrative genomic analyses, genome-wide genetic perturbation screens in patient-derived GBM models and independent lines of validation. Specific genetic dependencies were found consistent across recurrent tumor models, accompanied by increased mutational burden and differential transcript and protein expression compared to its primary GBM predecessor. Our observations suggest a multi-layered genetic response to drive tumor recurrence and implicate PTP4A2 (protein tyrosine phosphatase 4A2) as a modulator of self-renewal, proliferation and tumorigenicity in recurrent GBM. Genetic perturbation or small-molecule inhibition of PTP4A2 acts through a dephosphorylation axis with roundabout guidance receptor 1 (ROBO1) and its downstream molecular players, exploiting a functional dependency on ROBO signaling. Because a pan-PTP4A inhibitor was limited by poor penetrance across the blood–brain barrier in vivo, we engineered a second-generation chimeric antigen receptor (CAR) T cell therapy against ROBO1, a cell surface receptor enriched across recurrent GBM specimens. A single dose of ROBO1-targeted CAR T cells doubled median survival in cell-line-derived xenograft (CDX) models of recurrent GBM. Moreover, in CDX models of adult lung-to-brain metastases and pediatric relapsed medulloblastoma, ROBO1 CAR T cells eradicated tumors in 50–100% of mice. Our study identifies a promising multi-targetable PTP4A–ROBO1 signaling axis that drives tumorigenicity in recurrent GBM, with potential in other malignant brain tumors. Functional CRISPR screens in patient-matched pre-treatment and post-treatment glioblastoma models identify the PTP4A–ROBO1 axis as a driver of tumorigenicity and enriched ROBO1 expression in recurrent glioblastoma that can be targeted with CAR T cell therapy.
BigField GEG Tech's insight:

With existing treatments such as surgery, radiotherapy, and chemotherapy, brain tumors often recur, and patients' survival is limited to just a few months. However, a team of researchers has discovered a new pathway cancer cells use to infiltrate the brain. To find this pathway, the researchers used large-scale gene-editing technology to compare genetic dependencies, particularly in glioblastoma at initial diagnosis and after its recurrence following standard treatments. This new pathway is used for axonal guidance, a signaling axis that helps establish standard brain architecture, which can be invaded by cancer cells. Their research also reveals that blocking this pathway using CAR-T cells, mainly targeting the Roundabout Guidance Receptor 1 protein, looks promising for stopping and killing these tumors. The treatment was tested in mouse models on three different types of cancer: adult glioblastoma, adult lung-cerebral metastases, and pediatric medulloblastoma. In all three models, the treatment doubled survival time. In two of the three diseases, the tumor was eradicated in at least 50% of the mice. 

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CD22-targeted CAR-T therapy shows promising results in relapsed large B-cell lymphoma

CD22-targeted CAR-T therapy shows promising results in relapsed large B-cell lymphoma | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
CAR-T cell therapy, which targets a specific protein on the surface of cancer cells, causes tumors to shrink or disappear in about half of patients with large B-cell lymphoma who haven't experienced improvement with chemotherapy treatments.
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One of CAR-T cell therapy targets CD19 on the surface of lymphoma cells. It causes tumors to shrink or disappear in around half of patients with large B-cell lymphoma who have not improved with chemotherapy treatments. If this CAR-T treatment fails or the cancer recurs, as it does in around half of patients, the prognosis is dire. The median survival time after a relapse is around six months. However, a phase 1 clinical trial conducted at Stanford Medicine showed that a new CAR-T cell therapy targeting CD22 on the surface of cancer cells significantly improved outcomes for these patients: more than half of the 38 people enrolled in the trial, 37 of whom had already relapsed after initial CAR-T therapy, experienced a complete response to their cancer. Over half of the patients treated lived for at least two years after treatment.

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Brainwide silencing of prion protein by AAV-mediated delivery of an engineered compact epigenetic editor

Brainwide silencing of prion protein by AAV-mediated delivery of an engineered compact epigenetic editor | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Prion disease is caused by misfolding of the prion protein (PrP) into pathogenic self-propagating conformations, leading to rapid-onset dementia and death. However, elimination of endogenous PrP halts prion disease progression. In this study, we describ
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Prion disease, which leads to rapid neurodegeneration and death, is caused by the presence of deformed versions of the prion protein. These cause a cascade effect in the brain: the defective prion proteins deform other proteins, and together these proteins not only cease to function correctly but also form toxic aggregates that kill neurons—most conventional drugs work by targeting just one protein. However, in less than two years, using a research tool called CRISPRoff, researchers have developed a set of molecular tools called CHARMs that act upstream, disabling the gene that codes for a defective protein so that the protein is never produced. CHARMs achieve this by epigenetic editing. Unlike gene editing, epigenetic editing does not alter the underlying DNA, and the gene itself remains intact. Furthermore, like gene editing, epigenetic editing is stable, meaning that a gene switched off by CHARM should remain switched off. This would mean that patients would only have to take CHARM once, unlike other drugs. Moreover, to address the transmission problem, the researchers have also designed an AAV vector that can enter the brain more efficiently by exploiting a pathway that naturally transports iron in the brain.  

 

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Strong, flexible ‘nylon’ made by engineered bacteria for the first time - Nature

Strong, flexible ‘nylon’ made by engineered bacteria for the first time - Nature | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
The bioplastic was malleable, but is more expensive to produce than are plastics made from fossil fuels.
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Researchers have genetically engineered microbes to produce a strong, flexible plastic similar to nylon for the first time.

Bacteria have been used to generate polyesters such as polyhydroxyalkanoates (PHAs) in the past but, nylon-like plastics such as those used in clothing and shoe manufacturing have been difficult to create, the authors report in Nature Chemical Biology 

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An ancient RNA-guided system could simplify delivery of gene editing therapies | Broad Institute

An ancient RNA-guided system could simplify delivery of gene editing therapies | Broad Institute | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
The programmable proteins are compact, modular, and can be directed to modify DNA in human cells.
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A vast search of natural diversity has led scientists at MIT’s McGovern Institute and the Broad Institute of MIT and Harvard to uncover ancient systems with the potential to expand the genome editing toolbox. 

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Functional characterization of variants improves cancer risk assessment and treatment

Functional characterization of variants improves cancer risk assessment and treatment | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Findings from a multi-institutional, international study led by researchers from the Mayo Clinic Comprehensive Cancer Center have significantly advanced the understanding of genetic alterations in the BRCA2 gene, a key player in hereditary cancer risk.
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The results of an international multi-institutional study have significantly improved our understanding of genetic alterations in the BRCA2 gene, a key player in hereditary cancer risk. Researchers have carried out a comprehensive functional assessment of all possible variants in the crucial DNA-binding domain of the BRCA2 gene, resulting in the clinical classification of 91% of variants of uncertain significance (VUS) in this part of the gene. This discovery significantly improves the accuracy of genetic testing, and will enable healthcare professionals to offer more accurate risk assessments and personalized treatment plans for people carrying these variants. The study, published in Nature , used CRISPR-Cas9 gene-editing technology to analyze the functional impact of nearly 7,000 BRCA2 gene variants, definitively identifying those that increase cancer risk and those that do not. This new information will remove much of the uncertainty surrounding VUS, enabling more informed decisions to be made about cancer screening, preventative measures and treatment strategies. 

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Genome editing with the HDR-enhancing DNA-PKcs inhibitor AZD7648 causes large-scale genomic alterations | Nature Biotechnology

Genome editing with the HDR-enhancing DNA-PKcs inhibitor AZD7648 causes large-scale genomic alterations | Nature Biotechnology | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
The DNA-PKcs inhibitor AZD7648 enhances CRISPR–Cas9-directed homology-directed repair efficiencies, with potential for clinical utility, but its possible on-target consequences are unknown. We found that genome editing with AZD7648 causes frequent kilobase-scale and megabase-scale deletions, chromosome arm loss and translocations. These large-scale chromosomal alterations evade detection through typical genome editing assays, prompting caution in deploying AZD7648 and reinforcing the need to investigate multiple types of potential editing outcomes. A compound that enhances homology-directed repair in CRISPR editing leads to genome instability.
BigField GEG Tech's insight:

The CRISPR-Cas9 molecular complex is the tool most widely used by scientists around the world. It cuts double-stranded DNA at the exact point where the genetic material is to be modified. The cut activates two natural repair mechanisms, including homology-directed repair. To induce the cell to use this repair, researchers have recently begun using a molecule called AZD7648, which blocks rapid repair and forces the cell to use homology-directed repair. This approach should accelerate the development of more effective gene therapies. Initial studies on these new approaches have been positive. However, a research group has just discovered that the use of AZD7648 has serious side effects. The modifications would result in the outright deletion of thousands of DNA building blocks, known as bases. Entire chromosome arms were even detached, rendering the genome unstable, with unpredictable consequences for the cells modified by this technique. The study has just been published in the journal Nature Biotechnology. 

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Engineered SNIPRs transform CAR T-cell precision for safer cancer therapy

Engineered SNIPRs transform CAR T-cell precision for safer cancer therapy | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Researchers developed SNIPRs, innovative receptors that sense soluble ligands, enabling precise therapeutic control in CAR T-cells. This breakthrough enhances tumor targeting while reducing off-target effects, paving the way for advanced synthetic biology applications.
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A Nature study introduces Synthetic Intramembrane Proteolysis Receptors (SNIPRs) to improve the precision and safety of CAR-T cell therapies to target solid tumors. This novel receptor system enables engineered cells to detect soluble ligands with high sensitivity and specificity, reducing the risk of off-target effects that often cause severe side effects in conventional therapies. In preclinical models, SNIPR-equipped CAR-T cells demonstrated superior tumor specificity and minimized damage to healthy tissues. This advancement represents a pivotal step toward safer and more effective cellular immunotherapies, particularly for cancers with challenging antigen profiles. profiles.

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December 3, 2024 6:33 AM
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Large DNA deletions occur during DNA repair at 20-fold lower frequency for base editors and prime editors than for Cas9 nucleases | Nature Biomedical Engineering

Large DNA deletions occur during DNA repair at 20-fold lower frequency for base editors and prime editors than for Cas9 nucleases | Nature Biomedical Engineering | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
When used to edit genomes, Cas9 nucleases produce targeted double-strand breaks in DNA. Subsequent DNA-repair pathways can induce large genomic deletions (larger than 100 bp), which constrains the applicability of genome editing. Here we show that Cas9-mediated double-strand breaks induce large deletions at varying frequencies in cancer cell lines, human embryonic stem cells and human primary T cells, and that most deletions are produced by two repair pathways: end resection and DNA-polymerase theta-mediated end joining. These findings required the optimization of long-range amplicon sequencing, the development of a k-mer alignment algorithm for the simultaneous analysis of large DNA deletions and small DNA alterations, and the use of CRISPR-interference screening. Despite leveraging mutated Cas9 nickases that produce single-strand breaks, base editors and prime editors also generated large deletions, yet at approximately 20-fold lower frequency than Cas9. We provide strategies for the mitigation of such deletions. DNA repair after Cas9-mediated double-strand breaks induces large DNA deletions at frequencies 20-fold higher than elicited by base editors and prime editors leveraging Cas9 nickases producing single-strand breaks.
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In a paper recently published  in Nature Biomedical Engineering, Korean researchers report that large DNA deletions (>100 bp) occur during DNA repair at a much lower frequency for core and major editors than for Cas9 nucleases. The team performed in vitro gene editing on a panel of target sites in various cancer cell lines as well as in human embryonic stem cells and primary human T lymphocytes from two donors. DNA deletions were detected using a combination of CRISPR interference screening and an optimised long-range amplicon sequencing workflow that required the development of a k-mer alignment algorithm to simultaneously analyse large DNA deletions and small indels with high accuracy. The team found that Cas9-mediated double-strand breaks induced large deletions at variable frequencies ranging from 0.2% to 17.5% in all six cell lines, excluding primary T lymphocytes, in which the frequency of large deletions was much higher (around 15% on average for both donors).

 

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November 7, 2024 6:46 AM
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Innovative immunotherapy harnesses natural killer T cells to combat solid tumors

Innovative immunotherapy harnesses natural killer T cells to combat solid tumors | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
In the fight against cancer, chimeric antigen receptor T cell (CAR-T) therapy has achieved notable success in treating blood cancers. However, it has been largely ineffective against solid tumors.
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In the fight against cancer, CAR-T cells have achieved remarkable results in treating blood cancers. However, they have proven largely ineffective against solid tumors. CAR-NKT cells, on the other hand, possess innate properties that make them particularly well-equipped to combat solid tumors. Specifically, they express a T cell receptor that recognizes glycolipid antigens presented by CD1d molecules. CAR-NKT cells effectively eliminate CD1d-expressing M2 macrophages in the tumor microenvironment. These macrophages promote tumor growth and suppress immune responses, making their elimination essential for strengthening antitumor immunity. By targeting these cells, CAR-NKT therapy effectively reprograms the tumor microenvironment from a pro-tumor to an anti-tumor state. CAR-NKT cells promote epitope spreading, a process by which the immune system recognizes and attacks new targets, activating T-cell responses. Researchers further demonstrated that combining CAR-NKT cells with PD1 blockade significantly enhances their antitumor activity, and that this activity can be further strengthened by vaccination approaches, such as those using dendritic cells loaded with alpha-galactosylceramide. 

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October 28, 2024 11:13 AM
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RNA Repeat Targeting as Potential New Therapeutic Strategy for ALS and Frontotemporal Dementia

RNA Repeat Targeting as Potential New Therapeutic Strategy for ALS and Frontotemporal Dementia | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Researchers in the US report a potentially new therapeutic strategy for amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Using an optimised Cas13d-based RNA targeting system, they demonstrate significant in vitro and in vivo reduction of the RNA repeat mutations most commonly identified in patients with ALS or FTD.
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The hexanucleotide GGGGCC repeat expansion in the C9orf72 gene is the most common identified mutation in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) patients. This expansion leads to the production of repeat RNA and dipeptide repeat proteins (DPR), both of which contribute to disease and are recognised as important disease markers. In a recent study, a team in the United States demonstrated that CRISPR-Cas13d can be optimised to specifically target GGGGCC repeat RNA to minimise the translation of associated poly-dipeptides.The team shows that the optimised CRISPR-Cas13d system effectively reduced DPR production when delivered via lentiviral particles to various in vitro models, including C9orf72-ALS patient-derived induced pluripotent stem cells (iPSCs) and motor neurons derived from those iPSCs, and that the efficiency of RNA targeting corresponded to Cas13d expression levels. Intracerebroventricular injection of the C9orf72-targeting cargo (delivered via AAV9) in C9orf72 repeat transgenic mice revealed a significant reduction in DPR levels. The team reports that the approach exhibits specificity for expanded repeats, with minimal effect on wild-type C9orf72 transcripts and protein levels.

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October 8, 2024 9:48 AM
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World-first therapy using donor cells sends autoimmune diseases into remission

World-first therapy using donor cells sends autoimmune diseases into remission | Genetic Engineering Publications - GEG Tech top picks | Scoop.it

The treatment’s success in three people raises hopes for mass production of cutting-edge CAR T therapies.

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The treatment’s success in three people raises hopes for mass production of cutting-edge CAR T therapies.

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September 30, 2024 6:02 AM
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New discovery offers hope for overcoming CAR-T cell therapy resistance

New discovery offers hope for overcoming CAR-T cell therapy resistance | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Mayo Clinic researchers mined the molecular foundations of cancer and uncovered a new reason chimeric antigen receptor (CAR-T cell therapy) fails in some patients.
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CAR-T cell therapy works very well in only about a third of cancer patients. One of the main factors in failure is T-cell depletion. This condition causes many patients to relapse within a year of CAR-T cell therapy. In search of new solutions, a research team compared data from patients in remission with those whose CAR-T cell therapy had failed. They also studied how CAR-T cells killed tumors grown in laboratory mice. They compared the results of mice that responded well to CAR-T therapy with those that did not. The team observed an increase in interleukin-4 (IL-4) protein in both human and mouse CAR-T cell depletion samples. The team then used CRISPR gene-editing technology to eliminate or modify the IL-4 protein causing CAR-T cell dysfunction. The researchers also tested monoclonal antibodies to block or neutralize the IL-4 protein. They found that they also rejuvenated CAR-T cells and their ability to block cancer. 

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September 10, 2024 5:06 AM
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CD5 knockout using CRISPR boosts CAR T cell therapy efficacy

CD5 knockout using CRISPR boosts CAR T cell therapy efficacy | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
The effectiveness of CAR T cell therapy against a variety of cancers, including solid tumors, could be boosted greatly by using CRISPR-Cas9 technology to knock out the gene for CD5, a protein found on the surface of T cells, according to a preclinical study from investigators at the University of Pennsylvania's Perelman School of Medicine and Abramson Cancer Center.
BigField GEG Tech's insight:

CAR T cells are T cells designed to attack specific targets on cancer cells. They have produced remarkable results in certain patients with blood cancers. However, they have not worked well against other cancers, particularly solid tumor cancers such as pancreatic cancer, prostate cancer, and melanoma. The researchers looked for techniques to increase the effectiveness of CAR T cell therapy. The study, published today in Science Immunology, suggests that using CRISPR-Cas9 technology to knock out the CD5 gene could be a first-rate technique. By shedding light on the hitherto obscure role of the CD5 protein present on the surface of T cells, the researchers discovered that it functions as a robust immune checkpoint, limiting the efficiency of T lymphocytes. By deleting it, they showed that the anti-cancer activity of CAR T cells improved considerably in various preclinical cancer models. A Phase I clinical trial of CD5-knockout CAR T cells will soon begin enrolling patients with CD5-bearing T-cell lymphoma.

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September 6, 2024 3:50 AM
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Cellular immunotherapy offers hope for solid tumors

Cellular immunotherapy offers hope for solid tumors | Genetic Engineering Publications - GEG Tech top picks | Scoop.it
Early findings from a small clinical trial provide evidence that a new cellular immunotherapy approach may be effective in treating metastatic solid tumors. In the trial, researchers from the National Institutes of Health (NIH) genetically engineered normal white blood cells, known as lymphocytes, from each patient to produce receptors that recognize and attack their specific cancer cells.
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To date, no effective cell therapy for solid cancers has been found. However, initial results from a small clinical trial show that a new cellular immunotherapy approach could effectively treat metastatic solid tumors. Researchers collected lymphocytes from each patient's tumor as part of the trial. They then used sophisticated molecular characterization techniques to identify and isolate the receptors on these lymphocytes, called T-cell receptors, which recognized specific changes in each patient's tumor. After genetically sequencing these receptors, they used a retrovirus to insert the receptor genes into normal lymphocytes taken from each patient's circulating blood. The initial results came from people with metastatic colorectal cancer who had already undergone several previous treatments. Personalized immunotherapy shrank tumors in several patients and prevented tumors from growing back for up to seven months. The results were published on 11 July 2024 in Nature Medicine.

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