Metabolic reprogramming is linked to cancer cell growth and proliferation, metastasis, and therapeutic resistance in a multitude of cancers. Targeting dysregulated metabolic pathways to overcome resistance, an urgent clinical need in all relapsed/refractory cancers, remains difficult. Through genomic analyses of clinical specimens, we show that metabolic reprogramming toward oxidative phosphorylation (OXPHOS) and glutaminolysis is associated with therapeutic resistance to the Bruton’s tyrosine kinase inhibitor ibrutinib in mantle cell lymphoma (MCL), a B cell lymphoma subtype with poor clinical outcomes. Inhibition of OXPHOS with a clinically applicable small molecule, IACS-010759, which targets complex I of the mitochondrial electron transport chain, results in marked growth inhibition in vitro and in vivo in ibrutinib-resistant patient-derived cancer models. This work suggests that targeting metabolic pathways to subvert therapeutic resistance is a clinically viable approach to treat highly refractory malignancies.

Kidney failure is common in patients with Coronavirus Disease-19 (COVID-19) resulting in increased morbidity and mortality. In an international collaboration, 284 kidney biopsies were evaluated to improve understanding of kidney disease in COVID-19. Diagnoses were compared to five years of 63,575 native biopsies prior to the pandemic and 13,955 allograft biopsies to identify diseases increased in patients with COVID-19. Genotyping for APOL1 G1 and G2 alleles was performed in 107 African American and Hispanic patients. Immunohistochemistry for SARS-CoV-2 was utilized to assess direct viral infection in 273 cases along with clinical information at the time of biopsy. The leading indication for native biopsy was acute kidney injury (45.4%), followed by proteinuria with or without concurrent acute kidney injury (42.6%). There were more African American patients (44.6%) than patients of other ethnicities. The most common diagnosis in native biopsies was collapsing glomerulopathy (25.8%) which associated with high-risk APOL1 genotypes in 91.7% of cases. Compared to the five-year biopsy database, the frequency of myoglobin cast nephropathy and proliferative glomerulonephritis with monoclonal IgG deposits was also increased in patients with COVID-19 (3.3% and 1.7%, respectively), while there was a reduced frequency of chronic conditions (including diabetes mellitus, IgA nephropathy, and arterionephrosclerosis) as the primary diagnosis. In transplants, the leading indication was acute kidney injury (86.4%), for which rejection was the predominant diagnosis (61.4%). Direct SARS-CoV-2 viral infection was not identified. Thus, our multi-center large case series identified kidney diseases that disproportionately affect patients with COVID-19, demonstrated a high frequency of APOL1 high-risk genotypes within this group, with no evidence of direct viral infection within the kidney.

A rat brain cDNA library was screened by using a mixture of oligonucleotides whose sequences were deduced from the amino acid sequence of a human placental protein-tyrosine-phosphatase (PTPase; EC 3.1.3.48) reported by Charbonneau et al. [Charbonneau, H., The levels of phosphotyrosine found in the proteins of eukaryotic cells are tightly controlled by the opposing actions of protein-tyrosine kinases and protein-tyrosine-phosphatases (PTPases; EC 3.1.3.48). Increases in phosphotyrosine due to the activation of protein-tyrosine kinases are associated with the responses of cells to numerous external stimuli, most notably stimuli that trigger cell proliferation (for reviews see refs. 1 and 2). Under normal conditions, such phosphorylations are transient and are reversed by the actions of cellular PTPases. If the dephosphorylation of phosphotyrosine is prevented by the use of PTPase inhibitors (e.g., orthovanadate), cellular phosphotyrosine increases and certain cells exhibit a transformed phenotype (3). These studies indicate the importance of PTPases to the maintenance of the normal growth properties of cells.Substantial evidence is accumulating to indicate that PTPases constitute a family of important enzymes. Tonks et al. (4,5) recently reported the isolation of a 37-kDa protein from human placenta having PTPase activity. The amino acid sequence of this protein showed sequence similarity to the cytoplasmic domain of a protein found in the immune system known as CD45 (6). CD45 was subsequently shown to have tyrosine-phosphatase activity (7). Streuli et al. (8) MATERIALS AND METHODSMaterials. Restriction endonucleases and modifying enzymes were purchased from either New England Biolabs or United States Biochemical. The rat hypothalamic cDNA library, constructed in a pCD vector, was kindly provided by M. Brownstein (National Institutes of Health). The E. coli expression system using T7 RNA polymerase was provided by S. Tabor (Harvard Medical School).Screening of the cDNA Library. A rat hypothalamic cDNA library was plated at a density of 30,000 colonies per 150-mm plate. Colonies were transferred to nitrocellulose filters and then amplified overnight on chloramphenicol-containing plates (10). A total of 540,000 recombinants were initially screened by the method of Grunstein and Hogness (11), using a 32P-labeled synthetic oligonucleotide probe, d(TTCTGYT-CCCANACCATYTCCCARAA) (Y = pyrimidine, R = purine, N = any nucleoside). The sequence of this oligonucleotide was designed on the basis of the published amino acid sequence (Phe-Trp-Glu-Met-Val-Trp-Glu-Gln-Lys) of human placental PTPase-1B (6). Hybridization was carried out for 44 hr at 55°C in 6x SSC (lx SSC is 150 mM NaCl/15 mM sodium citrate)/0.1 sodium pyrophosphate/0.2% SDS containing heparin at 200 jig/ml and radiolabeled probe at 1 x 107 cpm per filter. The filters were washed four times at 55°C for 30 min in 6x SSC/0.1% SDS and then subjected to autoradiography with an intensifying screen.DNA Sequence Analysis, Northern Hybridization, and in Situ Lo...

Poly(A) RNA was isolated from a rat medullary thyroid carcinoma that exhibited high levels of immunoreactive cholecystokinin (CCK). Double-stranded cDNA was synthesized from the poly(A) RNA and inserted into the Pst I site of pBR322. Bacterial colonies containing CCK cDNA were identified using the hybridization probe d(T-C-C-A-T-C-C-A-N-C-C-C-A-T-G-T-A-G-T-C). The sequence of the probe was deduced from the known amino acid sequence of porcine CCK-8, Asp-Tyr-Met-Gly-Trp-Met-Asp-Phe-NH2. The nucleotide sequence of the cDNA complementary to the mRNA of rat preprocholecystokinin was determined. The cDNA contains 33 nucleotides in the 5'-noncoding region, 199 nucleotides in the 3'-noncoding region, and 345 nucleotides coding for a precursor to CCK, which is 115 amino acids (Mr, 12,826). Examination of the rat CCK gene revealed a suggested transcriptional control sequence analogous to the "TATA" sequence located 33 nucleotides upstream from a proposed transcriptional start site. The amino acid sequence of CCK-39 is flanked by both amino-terminal and carboxyl-terminal extensions. Analysis of CCK mRNA showed that it is approximately equal to 750 nucleotides long. CCK mRNA of the rat brain and intestine appeared to be identical in size to the CCK mRNA of the carcinoma.