Spinigerin induces apoptotic like cell death in a caspase independent manner in Leishmania donovani

Sardar, Abul Hasan; Das, Sushmita; Agnihorti, S.; Kumar, Maneesh; Ghosh, Ayan Kr.; Abhishek, Kumar; Kumar, Ashish; Purkait, Bidyut; Ansari, Yousuf; Das, Pradeep

doi:10.1016/j.exppara.2013.10.011

Cited by 26 publications

(20 citation statements)

References 38 publications

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“…8B). Similar observations were reported by Sardar et al, who showed an increase in lipid peroxide level in L. donovani after spinigerin treatment (30).…”

Section: Amphotericin B Drug Resistance In Leishmania Donovanisupporting

confidence: 79%

In Vitro Evaluation of Antileishmanial Activity of Computationally Screened Compounds against Ascorbate Peroxidase To Combat Amphotericin B Drug Resistance

Mansuri

Kumar

Rana

et al. 2017

Antimicrob Agents Chemother

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In visceral leishmaniasis (VL), the host macrophages generate oxidative stress to destroy the pathogen, while Leishmania combats the harmful effect of radicals by redox homeostasis through its unique trypanothione cascade. Leishmania donovani ascorbate peroxidase (LdAPx) is a redox enzyme that regulates the trypanothione cascade and detoxifies the effect of H 2 O 2 . The absence of an LdAPx homologue in humans makes it an excellent drug target. In this study, the homology model of LdAPx was built, including heme, and diverse compounds were prefiltered (PAINS, ADMET, and Lipinski's rule of five) and thereafter screened against the LdAPx model. Compounds having good affinity in terms of the Glide XP (extra precision) score were clustered to select diverse compounds for experimental validation. A total of 26 cluster representatives were procured and tested on promastigote culture, yielding 12 compounds with good antileishmanial activity. Out of them, six compounds were safer on the BALB/c peritoneal macrophages and were also effective against disease-causing intracellular amastigotes. Three out of six compounds inhibited recombinant LdAPx in a noncompetitive manner and also demonstrated partial reversion of the resistance property in an amphotericin B (AmB)-resistant strain, which may be due to an increased level of reactive oxygen species (ROS) and decrease of glutathione (GSH) content. However, inhibition of LdAPx in resistant parasites enhanced annexin V staining and activation of metacaspase-like protease activity, which may help in DNA fragmentation and apoptosis-like cell death. Thus, the present study will help in the search for specific hits and templates of potential therapeutic interest and therefore may facilitate the development of new drugs for combination therapy against VL.KEYWORDS ascorbate peroxidase, phylogenetic analysis, virtual screening and docking, apoptosis, inhibitors, leishmania, enzymatic assay V isceral leishmaniasis (VL) is one of the fatal neglected parasitic diseases and is caused by the flagellated protozoan parasite of the genus Leishmania and family Trypanosomatidae (1). The promastigote forms are transmitted to the host blood through the bite of phlebotomine sand flies (vector) and are differentiated into amastigotes in macrophages, resulting in hepatosplenomegaly (2). According to a WHO report from 2015, approximately 30,000 annual deaths and over 1.3 million new cases were estimated. The countries most affected by VL were noted as Bangladesh, Brazil,

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“…8B). Similar observations were reported by Sardar et al, who showed an increase in lipid peroxide level in L. donovani after spinigerin treatment (30).…”

Section: Amphotericin B Drug Resistance In Leishmania Donovanisupporting

confidence: 79%

In Vitro Evaluation of Antileishmanial Activity of Computationally Screened Compounds against Ascorbate Peroxidase To Combat Amphotericin B Drug Resistance

Mansuri

Kumar

Rana

et al. 2017

Antimicrob Agents Chemother

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“…The two enzymes overexpressed very highly in the early hours of differentiation in amastigotes (Fig. 5B), which demonstrated that PPP is early responsive and aids the parasite to cope with the initial phase of oxidative stress and their expression subsides as the parasite establishes infection in the form of amastigote (36). The phenomenon was found to be unrelated to glucose content of the axenic media as the expression of Leishmania GT2 was found to be unaltered and glucose uptake was not drastically changed (Supplemental Fig.…”

Section: Discussionmentioning

confidence: 99%

“…It can be proposed that an operative circle may be playing between ascorbate-dependent peroxidase, trypanothione reductase, and PPP enzymes to complete the trypanothione reduction machinery as trypanosomatids lack a selenium containing glutathione based peroxidase-reductase system (8). Lipid peroxidation in Ld caused caspase-independent apoptotic cell death when treated with spinigerin, an antimicrobial peptide proposed to have antileishmanial effect through generation of ROS (36).…”

Section: Discussionmentioning

confidence: 99%

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Metabolic reconfiguration of the central glucose metabolism: a crucial strategy ofLeishmania donovanifor its survival during oxidative stress

et al. 2015

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Understanding the mechanism that allows the intracellular protozoan parasite Leishmania donovani (Ld) to respond to reactive oxygen species (ROS) is of increasing therapeutic importance because of the continuing resistance toward antileishmanial drugs and for determining the illusive survival strategy of these parasites. A shift in primary carbon metabolism is the fastest response to oxidative stress. A 14 CO 2 evolution study, expression of glucose transporters together with consumption assays, indicated a shift in metabolic flux of the parasites from glycolysis toward pentose phosphate pathway (PPP) when exposed to different oxidants in vitro/ex vivo. Changes in gene expression, protein levels, and enzyme activities all pointed to a metabolic reconfiguration of the central glucose metabolism in response to oxidants. Generation of glucose-6-phosphate dehydrogenase (G6PDH) (∼5-fold) and transaldolase (TAL) (∼4.2-fold) overexpressing Ld cells reaffirmed that lethal doses of ROS were counterbalanced by effective manipulation of NADPH:NADP + ratio and stringent maintenance of reduced thiol content. The extent of protein carbonylation and accumulation of lipid peroxidized products were also found to be less in overexpressed cell lines. Interestingly, the LD 50 of sodium antimony gluconate (SAG), amphotericin-B (AmB), and miltefosine were significantly high toward overexpressing parasites. Consequently, this study illustrates that Ld strategizes a metabolic reconfiguration for replenishment of NADPH pool to encounter oxidative

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“…Intracellular Calcium concentration was measured by using the fluorescent probe Fura-2-AM (Sigma Aldrich, USA) as described previously by Sardar et al , 2013 [29]. Ethylene glycol-bis(2-aminoethylether)-N,N,N’,N’-tetraacetic acid (EGTA), a calcium chelator was used as control.…”

Section: Methodsmentioning

confidence: 99%

Intracellular zinc flux causes reactive oxygen species mediated mitochondrial dysfunction leading to cell death in Leishmania donovani

et al. 2017

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Leishmaniasis caused by Leishmania parasite is a global threat to public health and one of the most neglected tropical diseases. Therefore, the discovery of novel drug targets and effective drug is a major challenge and an important goal. Leishmania is an obligate intracellular parasite that alternates between sand fly and human host. To survive and establish infections, Leishmania parasites scavenge and internalize nutrients from the host. Nevertheless, host cells presents mechanism like nutrient restriction to inhibit microbial growth and control infection. Zinc is crucial for cellular growth and disruption in its homeostasis hinders growth and survival in many cells. However, little is known about the role of zinc in Leishmania growth and survival. In this study, the effect of zinc on the growth and survival of L.donovani was analyzed by both Zinc-depletion and Zinc-supplementation using Zinc-specific chelator N, N, N', N'–tetrakis (2-pyridylmethyl) ethylenediamine (TPEN) and Zinc Sulfate (ZnSO4). Treatment of parasites with TPEN rather than ZnSO4 had significantly affected the growth in a dose- and time-dependent manner. The pre-treatment of promastigotes with TPEN resulted into reduced host-parasite interaction as indicated by decreased association index. Zn depletion resulted into flux in intracellular labile Zn pool and increased in ROS generation correlated with decreased intracellular total thiol and retention of plasma membrane integrity without phosphatidylserine exposure in TPEN treated promastigotes. We also observed that TPEN-induced Zn depletion resulted into collapse of mitochondrial membrane potential which is associated with increase in cytosolic calcium and cytochrome-c. DNA fragmentation analysis showed increased DNA fragments in Zn-depleted cells. In summary, intracellular Zn depletion in the L. donovani promastigotes led to ROS-mediated caspase-independent mitochondrial dysfunction resulting into apoptosis-like cell death. Therefore, cellular zinc homeostasis in Leishmania can be explored for new drug targets and chemotherapeutics to control Leishmanial growth and disease progression.

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Spinigerin induces apoptotic like cell death in a caspase independent manner in Leishmania donovani

Cited by 26 publications

References 38 publications

In Vitro Evaluation of Antileishmanial Activity of Computationally Screened Compounds against Ascorbate Peroxidase To Combat Amphotericin B Drug Resistance

In Vitro Evaluation of Antileishmanial Activity of Computationally Screened Compounds against Ascorbate Peroxidase To Combat Amphotericin B Drug Resistance

Metabolic reconfiguration of the central glucose metabolism: a crucial strategy ofLeishmania donovanifor its survival during oxidative stress

Intracellular zinc flux causes reactive oxygen species mediated mitochondrial dysfunction leading to cell death in Leishmania donovani

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