Abstract Tumors of human non–small cell lung cancer (NSCLC) are heterogeneous but exhibit elevated glycolysis and glucose oxidation relative to benign tissues. Heme is a central molecule for oxidative metabolism ATP generation via mitochondrial phosphorylation (OXPHOS). Here, we showed that levels heme synthesis uptake, heme, oxygen-utilizing hemoproteins, oxygen consumption, generation, key biogenesis regulators were enhanced in NSCLC cells nontumorigenic cells. Likewise, proteins enzymes...

Abstract Lung cancer remains the leading cause of cancer-related death, despite advent targeted therapies and immunotherapies. Therefore, it is crucial to identify novel molecular features unique lung tumors. Here, we show that cyclopamine tartrate (CycT) strongly suppresses growth subcutaneously implanted non-small cell (NSCLC) xenografts nearly eradicated orthotopically NSCLC xenografts. CycT reduces heme synthesis degradation in cells oxygen consumption purified mitochondria. In...

The yeast Gis1 protein is a transcriptional regulator belonging to the JMJD2/KDM4 subfamily of demethylases that contain JmjC domain, which are highly conserved from humans. They have important functions in histone methylation, cellular signaling and tumorigenesis. Besides serving as cofactor many proteins, heme known directly regulate activities proteins ranging regulators potassium channels. Here, we report novel mechanism governing regulation demethylase activities. We found two modules,...

Heme is an essential nutritional, metabolic, and signaling molecule in living organisms. Pathogenic microbes extract heme from hosts to obtain metallonutrient, while fuels mitochondrial respiration ATP generation lung tumor cells. Here, we generated small heme-sequestering proteins (HeSPs) based on bacterial hemophores. These HeSPs contain neutral mutations the heme-binding pocket hybrid sequences hemophores of different bacteria. We showed that bind effectively extracted hemoglobin. They...

The KDM4 histone demethylase subfamily is constituted of yeast JmjC domain-containing proteins, such as Gis1, and human Gis1 orthologues, KDM4A/B/C. proteins have important functions in regulating chromatin structure gene expression response to metabolic nutritional stimuli. Heme acts a versatile signaling molecule regulate cellular diverse organisms ranging from bacteria humans. Here, using purified containing the JmjN/C domain, we showed that heme stimulates activity domains KDM4A Cas well...

Abstract Lung adenocarcinoma (ADC) and squamous cell carcinoma (SCC) are two most common subtypes of lung cancer. Here, to identify new, targetable molecular properties both subtypes, we monitored changes in the levels heme- oxidative phosphorylation (OXPHOS)-related proteins during tumorigenesis. Heme is a central molecule for metabolism ATP generation via OXPHOS. Notably, ADC SCC tumors can be induced genetically engineered KLLuc mouse model harboring G12D Kras mutation conditional Lkb1...

<div>Abstract<p>Tumors of human non–small cell lung cancer (NSCLC) are heterogeneous but exhibit elevated glycolysis and glucose oxidation relative to benign tissues. Heme is a central molecule for oxidative metabolism ATP generation via mitochondrial phosphorylation (OXPHOS). Here, we showed that levels heme synthesis uptake, heme, oxygen-utilizing hemoproteins, oxygen consumption, generation, key biogenesis regulators were enhanced in NSCLC cells nontumorigenic cells. Likewise,...

<div>Abstract<p>Multispectral optoacoustic tomography (MSOT) is an emerging noninvasive imaging modality that can detect real-time dynamic information about the tumor microenvironment in humans and animals. Oxygen enhanced (OE)-MSOT monitor vasculature oxygenation during disease development or therapy. Here, we used MSOT OE-MSOT to examine mice response of human non–small cell lung cancer (NSCLC) xenografts a new class antitumor drugs, heme-targeting agents heme-sequestering...

<p>Fig. S2 The levels of heme in subcellular organelles NSCLC cell lines. (A) Heme cytoplasm. (B) nucleus. (C) endoplasmic reticulum. (D) Golgi. (E) plasma membrane. (F) OXPHOS complex subunit cytochrome c (CYCS) are elevated cells. (G) COX4I1 (H) heme-containing enzyme cyclooxygenase (PTGS2) Data plotted as mean {plus minus} SD. For statistical analysis, the tumorigenic cells were compared to non-tumorigenic with a Welch 2-sample t-test.*, p-value, 0.05, **, p-value < 0.005.</p>

<p>Fig. S1 (A) The levels of the rate-limiting heme biosynthetic enzyme ALAS1 are elevated in NSCLC cells. (B) transporter SLC46A1 (C) degradation HMOX1 Data plotted as mean {plus minus} SD. For statistical analysis, tumorigenic cells were compared to non-tumorigenic with a Welch 2-sample t-test.*, p-value ,0.05, **, < 0.005.</p>

<p>Fig. S4. Proteins and enzymes relating to heme mitochondrial functions are upregulated in human NSCLC tissues relative control normal tissues. The examined proteins include the rate-limiting synthetic enzyme ALAS1 (A), transporter SLC48A1 (B), hemoprotein OXPHOS subunit cytochrome c (CYCS) (C), PTGS2 (D), regulator promoting biogenesis TFAM (E). Shown representative Montages 10X images of tumor tissue sections stained with DAPI or antibodies against indicated proteins. yellow...

<p>Fig. S3 (A) NSCLC cell lines exhibit a varying degree of migration capabilities. (B) invasion The images shown are cells that had migrated across Transwell inserts (in A) or crossed chambers coated with Corning Matrigel matrix and also passed B). At least three independent experiments were carried out for every condition. Data plotted as mean {plus minus} SD. For statistical analysis, the levels in compared to HCC Welch 2-sample t-test.*, p-value, 0.05, **, p-value < 0.005.</p>

<p>Fig. S5 (A) The quantified luminescence signals representing tumor volumes from mice bearing orthotopic H1299 xenografts treated with saline (control), and SA (50 mg/kg, I.V.) every 3 days (n = 6/group). Treatments started 4 after cell implantation. Data are plotted as mean {plus minus} SD. For statistical analysis, the levels in tumors were compared to untreated a Welch 2-sample t-test. difference between control was not statistically significant. (B) sequences of Y. pestis HasA...

<p>Fig. S7. The effect of HSP2 on oxygenation subcutaneous NSCLC A549-luc tumor xenografts after 3 weeks treatment (same tumors as shown in Fig. S1). (A) Representative images spatial distribution oxygen saturation [sO2 (O2)] the challenge and amplitude signal change (Î"sO2) for each group from breathing air to 100% oxygen. sO2 is calculated by subtracting average while sizes were: Control: 111 mm3; HSP2: 51.9 mm3. (B & C) Quantification (O2) Î"sO2 group. For statistical analysis,...

<p>Fig. S8. HSP2 and CycT do not affect iron metabolism hypoxia induction of HIF1�. (A) Iron stain did detect any difference between control treated tumor tissues. Shown are montages, 10X, 40X images control, CycT-treated tissue sections stained with Stain. The blue rectangles in 10X the regions shown images, respectively. Scale bar: montage, 1 mm for controls 500 µm tissues; 200 µm, 50 µm. (B) Representative IHC H1299 NSCLC graphs showing levels transferrin receptor TFRC HSP2,...

<p>Fig. S5. (A & B) The effect of HSP2 on subcutaneous NSCLC H1299-luc tumor xenografts after 1.5 weeks treatment (mid-point for shown in Fig. 2). (A) Photos excised tumors from mice treated with saline (control) or HSP2. (B) Average masses control and A. (C-E) Comparison sO2 Î"sO2 levels mid-point end-point HSP2-treated tumors. (C) Representative images spatial distribution oxygen saturation [sO2 (O2)] the challenge (D) Quantification (E) amplitude signal change (Î"sO2) breathing...

<p>Fig. S2. HSP2 and CycT did not significantly affect blood liver functions. (A) Average red cell count in mice bearing subcutaneous lung tumor xenografts treated with or without CycT. (B) hemoglobin levels (C) serum ALT (alanine transaminase) (D) ATP measured cells from livers excised HSP2, CycT, control saline. Data are plotted as mean {plus minus} SD. For statistical analysis, the were compared to untreated a Welch 2-sample t-test.</p>

<p>Fig. S6 Analysis of correlation between tumor sizes and MSOT metrics. (A, B, & C) sO2 (Air), (O2), Î"sO2 as derived from data. No significant was detected any metric with measured ex vivo. (D) Summary p-values r correlations OE-MSOT metrics sizes. For statistical analysis, the levels in treated tumors were compared to control a Welch 2-sample t-test or Pearson correlation.</p>

<p>Fig. S1. HSP2 effectively suppresses the growth, oxygen consumption rates, and ATP generation in subcutaneous A549-luc NSCLC tumor xenografts. (A) Sizes of H1299 tumors (mm3) from mice treated with for 1.5 weeks (mid-point) or HSP2/CycT 3 (end-point). (B) Photos showing excised NOD SCID saline (control) (25 mg/kg I.V every days) (n=4/group). Average weights control shown A. (C) body bearing (D) Oxygen rates (OCRs) measured cells tumors. (E) levels Data are plotted as mean {plus...

<p>Fig. S3. Treatment with HSP2 or CycT does not affect mitochondrial structural proteins in tumor tissues. (A) Representative IHC images of H1299 NSCLC tissue sections and graphs showing the levels mitochondria marker TOMM20 control, HSP2, CycT-treated tumors. (B) VDAC1 Data are plotted as mean {plus minus} SD. For statistical analysis, treated mice were compared to untreated a Welch 2-sample t-test. No significance was detected any group.</p>

<p>Fig. S4. MSOT images from control and HSP2- or CycT-treated tumors of similar size. The were the same experiment shown in Fig. 2. quantified values for all parameters 2 not changed when tumor sizes matched. (A) spatial distribution baseline deoxyhemoglobin (Hb; blue), oxyhemoglobin (HbO2; red), total hemoglobin (THb). Maps constructed using back-projection spectral-unmixing overlaid on grayscale image corresponding animals each treatment group (control, HSP2 CycT). (B)...

<p>Fig. S8 The effect of HSP2 on the growth and progression H1299 NSCLC lung tumor xenografts blood liver functions in mice. (A) quantified luminescence signals representing volumes. Treatment started at week 3 when tumors have grown significantly BLI were about 5x107 photons/seconds. Data are plotted as mean {plus minus} SD. For statistical analysis, levels treated compared to untreated with a Welch 2-sample t-test. *, p-value < 0.05. (B) Representative H&E images control or...