CRISPR/Cas 9 Gene-Edited therapeutic / University of Florida, CRISPR Therap - LARVOL DELTA

[VIRTUAL] Nelfinavir Overcomes Proteasome Inhibitor Resistance in Multiple Myeloma By Modulating Membrane Lipid Bilayer Composition and Fluidity (ASH 2020) - "Nelfinavir is particularly effective in the treatment of proteasome inhibitor-refractory multiple myeloma (MM), where the combination of nelfinavir+bortezomib+dexamethasone yielded an overall response rate (ORR, PR or better) > 65% in a Phase II clinical trial...METHODS Proteome-wide affinity-purification of targets binding the nelfinavir active site was combined with genome-wide CRISPR/Cas9-based screening to identify protein partners interacting with nelfinavir and candidate genetic contributors affecting nelfinavir cytotoxicity...Pharmacologic targeting of membrane fluidity is a novel, potent mechanism to achieve anti-cancer activity, in particular against PI-refractory MM. This mechanism explains the clinical activity of nelfinavir in MM treatment as well as the key side effects of nelfinavir during antiretroviral therapy."

Hematological Malignancies • Human Immunodeficiency Virus • Infectious Disease • Metabolic Disorders • Multiple Myeloma • Oncology • ABCB1

Treatment with HIV-protease inhibitor nelfinavir identifies membrane lipid composition and fluidity as a therapeutic target in advanced multiple myeloma. (PubMed, Cancer Res) - "We combined proteome-wide affinity-purification of nelfinavir-interacting proteins with genome-wide CRISPR/Cas9-based screening to identify protein partners that interact with nelfinavir in an activity-dependent manner alongside candidate genetic contributors affecting nelfinavir cytotoxicity. Conversely, depletion of fatty acids/cholesterol pools by the FDA-approved drug ezetimibe showed a synergistic anti-cancer activity with nelfinavir in vitro. These results identify the modification of lipid-rich membranes by nelfinavir as a novel mechanism of action to achieve broad anti-cancer activity, which may be suitable for the treatment of PI-refractory multiple myeloma."

Journal • Hematological Malignancies • Human Immunodeficiency Virus • Infectious Disease • Multiple Myeloma • Oncology • ABCB1

Loss of function of a DMR6 ortholog in tomato confers broad-spectrum disease resistance. (PubMed, Proc Natl Acad Sci U S A) - "We then propose that SlDMR6 duplication in tomato resulted in subsequent subfunctionalization, in which SlDMR6-2 specialized in balancing SA levels in flowers/fruits, while SlDMR6-1 conserved the ability to fine-tune SA levels during pathogen infection of the plant vegetative tissues. Overall, this work not only corroborates a mechanism underlying SA homeostasis in plants, but also presents a promising strategy for engineering broad-spectrum and durable disease resistance in crops."

Journal • Infectious Disease

Biallelic editing of the LOB1 promoter via CRISPR/Cas9 creates canker-resistant 'Duncan' grapefruit. (PubMed, Phytopathology) - "No off-target mutations were detected in #DunYao7. This study represents the first time that CRISPR-mediated genome editing has been successfully used to generate disease-resistant plants for 'Duncan' grapefruit, paving the way for utilizing disease-resistant varieties to control canker."

Journal

Efficient genome editing using endogenous U6 snRNA promoter-driven CRISPR/Cas9 sgRNA in Sclerotinia sclerotiorum. (PubMed, Fungal Genet Biol) - "The U6-sgRNA system exhibited a significantly higher efficiency of genemutation at both loci. This technology provides a simple and efficient strategy for targeted gene mutation and thereby will accelerating the pace of research of pathogenicity and development in this economically important plant pathogen."

Journal

An N-terminal fusion allele to study melanin concentrating hormone receptor 1. (PubMed, Genesis) - "Using CRISPR/Cas9, we inserted the coding sequence of the fluorescent protein mCherry into the N-terminus of Mchr1...However, both lines should aid in studies of MCHR1 function and contribute to our understanding of MCHR1 signaling in the brain. Additionally, this approach could be expanded to aid in the study of other ciliary GPCRs."

Journal • GPRC6A

Identification and characterization of GLOBE, a major gene controlling fruit shape and impacting fruit size and marketability in tomato. (PubMed, Hortic Res) - "CRISPR/Cas9 knock out plants confirmed this gene as underlying the globe locus...Fruit measurements in three genetic backgrounds evidenced that globe impacts fruit size and several fruit shape attributes, pedicel length/width, and susceptibility of fruit to weather check. The mutant allele of GLOBE appears mostly recessive for all traits except fruit size where it acts additively."

Journal

Targeting histone acetylation dynamics and oncogenic transcription by catalytic P300/CBP inhibition. (PubMed, Mol Cell) - "CRISPR-Cas9 screening identified NCOR1 and HDAC3 transcriptional co-repressors as the principal antagonists of P300/CBP by counteracting acetylation turnover kinetics. Finally, deacetylation of H3K27 provides nucleation sites for reciprocal methylation switching, a feature that can be exploited therapeutically by concomitant KDM6A and P300/CBP inhibition. Overall, this study indicates that the steady-state histone acetylation-methylation equilibrium functions as a molecular rheostat governing cellular transcription that is amenable to therapeutic exploitation as an anti-cancer regimen."

Epigenetic controller • Journal • Hematological Disorders • Hematological Malignancies • Oncology • KDM6A • NCOR1

Simple, efficient and open-source CRISPR/Cas9 strategy for multi-site genome editing in Populus tremula × alba. (PubMed, Tree Physiol) - "A small deletion or a single nucleotide insertion at the single target site were the most common mutations. This CRISPR/Cas9 strategy arises as a rapid, simple, and standardized gene-editing tool to evaluate the gene role in essential developmental programs such as radial cell differentiation of poplar roots."

Journal

[VIRTUAL] A CRISPR/Cas9 Genome Wide Screen and Cryo-EM Structure Reveal Receptor Switching by a Synthetic AAV Variant (ASGCT 2021) - "Taken together, our results provide structure-function correlates for a newly evolved, synthetic AAV variant. Further, our approach underscores the potential for structure-guided evolution to improve AAV transduction through receptor switching and exploitation of cognate cell entry pathways independent of those employed by natural AAV serotypes."

Biomarker • Gastrointestinal Cancer • Gene Therapies • Hepatocellular Cancer • Liver Cancer • Solid Tumor

[VIRTUAL] A Novel GUCY2D(R838S) Knock-In Mouse Model of Autosomal Dominant Cone Rod Dystrophy (CORD6) Displays Progressive Photoreceptor Degeneration/ Dysfunction (ASGCT 2021) - "We have generated a novel R838S-KI mouse model with a genomic profile that mirrors human CORD6. Preliminary results reveal that R838S KI mice exhibit loss of rod structure and function, and aberrant cone function. This differs somewhat from disease progression in CORD6 patients."

Preclinical • Inherited Retinal Dystrophy • Ophthalmology • Retinitis Pigmentosa

Global Phosphoproteomics reveal CDK suppression as a vulnerability to KRAS addiction in Pancreatic Cancer. (PubMed, Clin Cancer Res) - "A link between CDK hyperactivation and mt KRas dependency was uncovered and pharmacologically-exploited to abrogate mt KRas-driven pancreatic cancer in highly relevant models, warranting clinical investigations of AT7519 in pancreatic cancer patients."

Clinical • Journal • CNS Disorders • Gastrointestinal Cancer • Hepatology • Oncology • Pancreatic Cancer • Psychiatry • Solid Tumor • CDK1 • CDK2 • CDK7 • CDK9 • KRAS

The SvFUL2 transcription factor is required for inflorescence determinacy and timely flowering in Setaria viridis. (PubMed, Plant Physiol) - "In S. viridis, perturbation of SvFul2, both through chemical mutagenesis and CRISPR/Cas9-based gene editing, converted a normally determinate inflorescence habit to an indeterminate one, and also repressed determinacy in axillary branch and floral meristems. Our analysis of gene networks connected to disruption of SvFul2 identified regulatory hubs at the intersect of floral transition and inflorescence determinacy, providing insights into the optimization of cereal crop architecture."

Journal • Immunology

The alternative initiation factor eIF2A plays key role in RAN translation of myotonic dystrophy type 2 CCUG•CAGG repeats. (PubMed, Hum Mol Genet) - "To better understand how DM2 LPAC and QAGR RAN proteins are expressed we generated a series of CRISPR/Cas9 edited HEK293T cell lines...For QAGR but not LPAC the eIF2A-dependent increases are not seen when p-eIF2α is blocked. These data highlight the differential regulation of DM2 RAN proteins and eIF2A as a potential therapeutic target for DM2 and other RAN diseases."

Journal • Genetic Disorders • Muscular Dystrophy • Myotonic Dystrophy • EIF2A

[VIRTUAL] Adenylate Kinase 2 Is a Selective Dependency in NSD2-High Multiple Myeloma (ASH 2020) - "Candidate genes were validated using CRISPR-Cas9 gene knockout and shRNA knockdown of individual target genes followed by in vitro competitive growth assays and cell viability assays...In addition, NSD2-high cells displayed higher sensitivity to the proteasome inhibitor bortezomib than NSD2-low cells suggesting elevated levels of endoplasmic reticulum (ER) stress in cells overexpressing NSD2...Conclusions : Our findings indicate that NSD2 high t(4;14) MM may have a vulnerability due to increased proteostatic stress. Accordingly, AK2 inhibition could be used in combination with proteasome inhibitors to treat MM patients with t (4;14) translocations by inducing the accumulation of lethal levels of unfolded proteins."

Hematological Malignancies • Multiple Myeloma • Oncology

[VIRTUAL] NSD2-E1099K Mutation Leads to Glucocorticoid-Resistant B Cell Lymphocytic Leukemia in Mice (ASH 2020) - "Using CRISPR/Cas9-edited isogenic ALL cell lines, we found that NSD2-E1099K mutation drove oncogenic programming by altering chromatin architecture, gene expression and enhancing cell growth, migration and infiltration to the central neural system (CNS)... The NSD2 mutation alters B cell development, particularly in an immunodeficient background and causes B cells to become resistant to glucocorticoids. The inability of the mutation to generate disease on its own except in an immunodeficient background suggests genes that collaborate with NSD2 in ALL may play a role in immune escape."

Acute Lymphocytic Leukemia • Hematological Disorders • Hematological Malignancies • Leukemia • Oncology • Pediatrics • Transplantation • CD19 • CD5 • ETV6 • RUNX1

[VIRTUAL] KDM6A Controls Genes Modulating Immune Surveillance in Multiple Myeloma (ASH 2020) - "We further modeled the loss of KDM6A in female MM cell lines using CRISPR-Cas9 ribonucleotide protein (RNP) technology and by re-expressing KDM6A in cells null for KDM6A... Together, our data demonstrates that KDM6A modulates EMT and directly regulates expression of major regulators of immune surveillance. KDM6A deficiency may drive MM towards an EMT program to promote systemic spread and facilitate escape from the immune surveillance."

Hematological Malignancies • Inflammatory Arthritis • Multiple Myeloma • Oncology • CCL3 • EZH2 • KDM6A • KMT2D • MLL2 • MLL3

[VIRTUAL] Nelfinavir overcomes proteasome inhibitor resistance in multiple myeloma by modulating the composition and fluidity of lipid membranes (DGHO 2020) - "Introduction: Treatment based on the HIV-protease inhibitor nelfinavir is active in > 65% patients with bortezomib-refractory multiple myeloma (MM)... Proteome-wide affinity-purification of targets binding the nelfinavir active site was combined with genome-wide CRISPR/Cas9-based screening to identify protein partners interacting with nelfinavir and candidate genetic contributors affecting nelfinavir cytotoxicity... We here identify pharmacologic targeting of membrane fluidity as a novel, potent mechanism to achieve anti-cancer activity, which is in particular suitable for the treatment of PI-refractory MM. This mechanism explains the clinical activity of nelfinavir in MM treatment as well as the key side effects of nelfinavir during antiretroviral therapy."

Hematological Malignancies • Infectious Disease • Multiple Myeloma • Oncology • ABCB1

[VIRTUAL] Nelfinavir overcomes proteasome inhibitor resistance in multiple myeloma by modulating the composition and fluidity of lipid membranes (DGHO 2020) - "Introduction: Treatment based on the HIV-protease inhibitor nelfinavir is active in > 65% patients with bortezomib-refractory multiple myeloma (MM)... Proteome-wide affinity-purification of targets binding the nelfinavir active site was combined with genome-wide CRISPR/Cas9-based screening to identify protein partners interacting with nelfinavir and candidate genetic contributors affecting nelfinavir cytotoxicity... We here identify pharmacologic targeting of membrane fluidity as a novel, potent mechanism to achieve anti-cancer activity, which is in particular suitable for the treatment of PI-refractory MM. This mechanism explains the clinical activity of nelfinavir in MM treatment as well as the key side effects of nelfinavir during antiretroviral therapy."

Hematological Malignancies • Infectious Disease • Multiple Myeloma • Oncology • ABCB1

[VIRTUAL] Nelfinavir overcomes proteasome inhibitor resistance in multiple myeloma by modulating the composition and fluidity of lipid membranes (DGHO 2020) - "Introduction: Treatment based on the HIV-protease inhibitor nelfinavir is active in > 65% patients with bortezomib-refractory multiple myeloma (MM)... Proteome-wide affinity-purification of targets binding the nelfinavir active site was combined with genome-wide CRISPR/Cas9-based screening to identify protein partners interacting with nelfinavir and candidate genetic contributors affecting nelfinavir cytotoxicity... We here identify pharmacologic targeting of membrane fluidity as a novel, potent mechanism to achieve anti-cancer activity, which is in particular suitable for the treatment of PI-refractory MM. This mechanism explains the clinical activity of nelfinavir in MM treatment as well as the key side effects of nelfinavir during antiretroviral therapy."

Hematological Malignancies • Infectious Disease • Multiple Myeloma • Oncology • ABCB1

[VIRTUAL] Nelfinavir overcomes proteasome inhibitor resistance in multiple myeloma by modulating the composition and fluidity of lipid membranes (DGHO 2020) - "Introduction: Treatment based on the HIV-protease inhibitor nelfinavir is active in > 65% patients with bortezomib-refractory multiple myeloma (MM)... Proteome-wide affinity-purification of targets binding the nelfinavir active site was combined with genome-wide CRISPR/Cas9-based screening to identify protein partners interacting with nelfinavir and candidate genetic contributors affecting nelfinavir cytotoxicity... We here identify pharmacologic targeting of membrane fluidity as a novel, potent mechanism to achieve anti-cancer activity, which is in particular suitable for the treatment of PI-refractory MM. This mechanism explains the clinical activity of nelfinavir in MM treatment as well as the key side effects of nelfinavir during antiretroviral therapy."

Hematological Malignancies • Infectious Disease • Multiple Myeloma • Oncology • ABCB1

[VIRTUAL] Unfolded Protein Response in CRISPR/Cas 9 Gene-Edited Pi*Z Alpha 1-Antitrypsin Hepatocytes and Pi*Z Alpha 1-Antitrypsin Transgenic Mouse Model (ATS-I 2020) - "This study provides new insights regarding PI*Z AAT elicited hepatic UPR. Our data improves understanding of underlying molecular pathological mechanisms of PI*Z AATD. These results may provide novel therapeutic targets for AATD."

Preclinical • Alpha-1 Antitrypsin Deficiency • AAT • PERK

Unfolded Protein Response in CRISPR/Cas 9 Gene-Edited Pi*Z Alpha 1-Antitrypsin Hepatocytes and Pi*Z Alpha 1-Antitrypsin Transgenic Mouse Model (ATS 2020) - "This study provides new insights regarding PI*Z AAT elicited hepatic UPR. Our data improves understanding of underlying molecular pathological mechanisms of PI*Z AATD. These results may provide novel therapeutic targets for AATD."

Preclinical • Alpha-1 Antitrypsin Deficiency • Genetic Disorders • Oncology • Solid Tumor • AAT